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PMC11276641_p20
|
PMC11276641
|
sec[2]/sec[0]/p[1]
|
3.1. Effects of Green Light Replacing Some Red and Blue Light on the Morphology and Photosynthesis of Melon Seedlings under Drought Stress
| 4.15625 |
biomedical
|
Study
|
[
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[
0.99951171875,
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The photosynthetic efficiency of plants is reduced under drought stress, and one of the main reasons is the decrease in gsw, which limits the gas exchange between plants and the outside world, resulting in lower uptake . The experimental results of this study showed that on the 3rd day of the initial drought period, stomatal conductance decreased significantly in the treatment with the addition of green light compared to the control, which was attributed to the fact that the addition of green light reversed the blue-light-mediated stomatal opening effect , resulting in a significant decrease in gsw, which led to a decrease in E in the melon seedlings. On the 6th day of treatment, Pn, gsw, and E were significantly higher in the treatment with added green light than in the CK treatment. It was hypothesized that melon seedlings under the CK treatment were significantly affected by drought stress, and they transpirated a large amount of water at the early stage of the treatment, so there was not enough water in the plants to maintain normal photosynthesis in the late stage of the treatment, whereas the group treated with added green light had a lower rate of transpiration at the early stage, and thus there was still enough water to maintain photosynthesis in the late stage.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p21
|
PMC11276641
|
sec[2]/sec[1]/p[0]
|
3.2. Effect of Green Light Replacing Some Red and Blue Light on Antioxidant System of Melon Seedlings under Drought Stress
| 4.195313 |
biomedical
|
Study
|
[
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A reactive oxygen species (ROS) plays a key role in plant response to abiotic stresses, and it mainly acts as a signaling molecule and plays an important regulatory role in plant adaptation to stress, but it also causes some degree of toxic effects to the plants . Under drought stress, this will lead to the production of and increase in ROS in plant tissues . ROS concentration will increase in cells and lead to the lipid peroxidation of membranes and may even lead to the oxidative damage of cells . When drought stress leads to the destruction of biofilm structure, MDA and electrolyte permeability can be used as characterization indicators to show the degree of damage it suffers . The results showed that on the 9th day of treatment, the green light replacing part of the red and blue light treatment showed a significant decrease in MDA and electrolyte osmolality compared with that under the control treatment, indicating that the addition of green light mitigated the degree of membrane lipid peroxidation caused by drought in melon seedlings. Compared with the CK treatment, the H 2 O 2 and O 2 ∙− content of the green light treatment showed a significant decrease, and the results of DAB and NBT staining of melon seedling leaves also confirmed this result, indicating that with the increase in the proportion of green light substitution, the ROS content of melon seedlings showed a decreasing trend.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p22
|
PMC11276641
|
sec[2]/sec[2]/p[0]
|
3.3. Effect of Green Light Replacing Some Red and Blue Light on Stomatal Morphology of Melon Seedlings under Drought Stress
| 4.136719 |
biomedical
|
Study
|
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Plants respond to various environmental stimuli through changes in defense cell volume expansion, which leads to stomatal opening and closing . Light is one of the most dynamic environmental signals and plays a key role in photosynthesis and stomatal movement , so light environmental conditions can be altered thereby changing stomatal opening to improve photosynthesis and water use efficiency . Red and blue light regulate stomatal movement through different regulatory pathways , and green light can reverse blue-light-induced stomatal opening . Therefore, green light can be used as a signal to control stomatal transpiration and plant water consumption . The experimental results showed that the addition of green light in the treatment had a significant effect on the stomatal pore length, pore width, and pore area of melon seedlings, and the pore area showed a significant decreasing trend as the proportion of green light replacement increased ( Table 2 ). It has been shown that exposing mature leaves to low light conditions triggers long-range signals that control stomatal development and leads to a decrease in stomatal density in young leaves . Ma et al. showed that stomatal density would increase with the increase in the percentage of green light . However, this phenomenon was not observed in this experiment, probably due to the fact that the plants were under drought stress, their growth and development were slow, and the treatment time was relatively short, so the phenomenon of stomatal density change was not observed.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p23
|
PMC11276641
|
sec[2]/sec[3]/p[0]
|
3.4. Effect of Green Light Percentage on Transcriptome of Phytohormones in Melon Seedlings under Drought Stress
| 4.441406 |
biomedical
|
Study
|
[
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In response to drought-induced oxidative damage, plants have evolved many defense strategies. Phytohormones respond in a highly synergistic manner to signals stimulated by the external environment, which is essential for plant growth and development , and are involved in multiple signaling pathways, which help plants to maintain a certain level of resilience under suboptimal conditions . In the untargeted metabolome assay for plants, the results showed large differences in ABA, JA, and SA in the two treatments, and the assay results for metabolites showed that the three phytohormones processed differential genes in three pathway nodes. In the ABA pathway, significant differential genes were seen in SnRK2 . Li et al. showed that SnRK2.6 interacts with phyB and is involved in light-induced stomatal opening and has a negative role in red-light-induced stomatal opening, and it has been shown that the ABA receptor interacts with the SnRK2-type kinase and inhibits the PP2C phosphatase activity, releases active SnRK2 kinase phosphorylation, and activates SLAC1 channels, leading to reduced guard cell swelling and stomatal closure , and so SnRK2 plays a key role in the ABA-regulated stomatal opening and closing pathway. In this experiment, the intensity of red and blue light in the light plasma was subsequently reduced as the proportion of green light substitution increased. The transcription factor MYC2 is an important regulator of the JA signaling pathway and plays a crucial role in the regulation of plant growth and stress tolerance. Under light/dark conditions, stomatal closure in the lower epidermis of MYC2-silenced leaves was weaker, water loss was faster, and drought tolerance was lower compared with that of the control plant leaves , and the study of poplar by Xia et al. The authors of showed that MYC2 regulates stomatal density and water use efficiency by targeting EPF2/EPFL4/EPFL9 and that overexpression of MYC2 reduces stomatal density and increases plant water use efficiency. It has been shown that the transcription factor MYC2 is involved in the development of Arabidopsis seedlings under blue light . The presence of a significantly up-regulated differential gene in MYC2 in this experiment indicated that stomatal opening was significantly affected by the JA pathway under different light-quality treatments. Previous studies have shown that MYC2 is involved in leaf stomatal development, and Ma et al. showed a significant change in stomatal density in a short-term drought treatment of cucumber seedlings in the treatment where green light replaced part of the red/blue light compared to the control treatment , but this phenomenon was not observed in this experiment. SA has been shown to ameliorate drought stress in plants by inducing stomatal closure in intact leaves and directly controlling stomatal movement by increasing the levels of ROS and nitric oxide (NO) in the guard cells , and it has been shown that TAG, with light-induced stomatal opening, is mainly mediated by PHOT1 and PHOT2 blue light receptors , and the results of this experiment showed that green light instead of partial blue light treatment resulted in the presence of a significantly up-regulated differential gene in TAG within the SA pathway, which enhanced the drought tolerance of melon seedlings. In addition to ABA, JA, and SA, T3 and CK treatments showed significant differential genes in multiple hormone signaling pathways of IAA, CK, GA, BR, and ETH , which play important regulatory roles in plant drought stress , and all are regulated by light signaling . Recently, it has been shown that green light is involved in the construction of plant photomorphology by regulating Arabidopsis hypocotyl elongation through BR . In this study, the partial replacement of red and blue light treatments by green light resulted in significant differential genes at several nodes of the BR pathway in melon seedlings, including BKI1, BAK1, and BSK, and it is hypothesized that BR hormones play an important role in the green-light-induced regulation of stomata to elicit resistance responses.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276641_p24
|
PMC11276641
|
sec[2]/sec[4]/p[0]
|
3.5. Effect of Green Light Percentage on Phytohormone Metabolome of Melon Seedlings under Drought Stress
| 4.328125 |
biomedical
|
Study
|
[
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0.0003986358642578125,
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[
0.9990234375,
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] |
Drought stress treatments have significant effects on phytohormone metabolism , and changes in the light environment have a greater impact on plant metabolism, and we focused on differential metabolites in the pathway of phytohormone biosynthesis . The results indicated that changes in the light environment mainly affected 13 differential metabolites. Among them, Aspartate, jasmonic acid, citric acid, Alpha-D-Glucose, tryptophan, and AMP differed significantly. When exposed to stress, plants accumulate large amounts of metabolites, especially amino acids. A series of studies have shown a close correlation between changes in Aspartate (Asp) content and plant stress . And, the degree of opening of plant stomata affects the accumulation of ASP, as changes in its level may also be related to malic acid activity . Ma et al. showed that increasing the proportion of green light in the plant’s light environment under drought stress affected the stomatal opening of cucumber seedlings and down-regulated the transcript levels of Aluminum-Activated Malate Transporter 9 (CsALMT9) , so the results of our experiments indicated that the addition of the green light treatment for JA is involved in a variety of abiotic stresses, including drought stress . JA pretreatment significantly increased drought tolerance in barley , and JA increased drought tolerance in cauliflower by activating the enzymatic antioxidant system . Our experimental results showed that JA content was significantly increased by adding green light treatment. Citric acid plays a key role in the initial stage of the TCA cycle. When present in excess, citric acid inhibits glycolysis, stimulates gluconeogenesis, and hinders downstream TCA reactions. Plants assume “dormancy” to prevent cells from wasting large amounts of energy and to ensure faster recovery under normal conditions . FRANCO et al. showed that citric acid accumulation correlates with plant photosynthesis and that at high PPFD, leaves of Clusia accumulated large amounts of malic and citric acids . Plants did not absorb green light, and the intensity of light (red and blue) effective for plant photosynthesis decreased under the T3 treatment compared to the control. Therefore, we found that the citric acid content was reduced under the T3 treatment compared to the CK group. Alpha D-glucose is a common reducing monosaccharide. The results of Lu et al. showed that the content of reducing sugars increased in rice under drought conditions and the content of reducing monosaccharides such as erythrulose, mannose, alpha D-glucose, and galactose increased in the metabolism group, and a variety of monosaccharides participated in the redox reactions of plants as cofactors and antioxidants. The results of this experiment showed a decrease in alpha D-glucose content under the T3 treatment compared to the CK treatment, indicating that the addition of green light reduced the degree of drought stress in plants . When plants are subjected to stress, they will accumulate more tryptophan, and as tryptophan is a target of oxidation, the free amino acid may provide a buffer between ROS and proteins in the chloroplast where both tryptophan and ROS are synthesized . Our results showed that the decrease in tryptophan content under the T3 treatment compared to the CK treatment could be attributed to the addition of green light reducing the stress level of seedlings, resulting in a decrease in the content of O 2 ∙− and H 2 O 2 , the main members participating in redox reactions. PAP has been reported to be a secondary messenger of ABA signaling that regulates the response to drought and oxidative stress , and AMP is a by-product of PAP that plays a role as a monomer in RNA production. It is a component of many metabolic processes . The results of this experiment showed a significant increase in AMP content under the CK treatment compared to the T3 treatment, indicating a more verified drought stress.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p25
|
PMC11276641
|
sec[3]/sec[0]/p[0]
|
4.1. Experimental Materials
| 3.902344 |
biomedical
|
Study
|
[
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This experiment was conducted in 2022 in the Biological and Environmental Engineering Laboratory of Facility Agriculture, College of Horticulture, Northwest A&F University. The test material was melon ‘Emerald’ ( Cucumis melo ), and the seeds were purchased from Shandong Zibo Harvest Seed Industry Science and Technology Co. Melon seeds were soaked in distilled water for 8 h and then germinated in the dark for 48 h. After germination, they were sown in 10 cm × 10 cm nutrient pots and grown in an artificial climatic chamber with a light intensity of 150 μmol/(m 2 ·s), day/night temperatures of 25 °C/20 °C, a photoperiod of 12 h/12 h, and a relative humidity of 60%, and were watered with 1/4 Hoagland melon nutrient solution (pH 6.5 ± 0.1, EC of 2.2–2.5 ms/cm) until the substrate reached water holding saturation.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p26
|
PMC11276641
|
sec[3]/sec[1]/p[0]
|
4.2. Experimental Design
| 3.515625 |
biomedical
|
Study
|
[
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[
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] |
After growing to 3 leaves and 1 heart for about 20 d in the artificial climate chamber, the seedlings with consistent growth were selected and transplanted into a cultivation frame equipped with an LED light source, in which the day and night temperatures were 28 °C/25 °C, relative humidity was 40%~50%, and drought stress was applied in the form of no watering, with a total of 9 d for the drought treatment.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276641_p27
|
PMC11276641
|
sec[3]/sec[1]/p[1]
|
4.2. Experimental Design
| 3.529297 |
biomedical
|
Study
|
[
0.9384765625,
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] |
[
0.9970703125,
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] |
We controlled the light quality through the LED light board; the LED light board consists of different light qualities of LED lamp beads, with a light board connected to the controller that can independently control the intensity of each light quality and light time. LED light panels provide a specific spectrum of light, and the quality, duration and intensity of the LED light is controlled through a controller.Optical panels and controllers manufactured by the Yinbian company (Xi’an, Shaanxi, China) Reflective films were posted all around the LED panels to ensure uniform radiation in all places. The PPFD of all treatments was controlled to be 250 μmol/(m 2 ·s), the red-to-blue ratio (R/B) was guaranteed to be 4:1 for all treatments, and the proportion of green light (G) was adjusted ( Table 3 ). The spectrogram under each treatment is shown in Figure 7 .
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p28
|
PMC11276641
|
sec[3]/sec[2]/sec[0]/p[0]
|
4.3.1. Morphometric Indicators
| 4.046875 |
biomedical
|
Study
|
[
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[
0.99853515625,
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0.0002371072769165039,
0.00004309415817260742
] |
After 9day of treatment, five plants were randomly selected and their aboveground height was determined using a scale; stem thickness was determined using vernier calipers; the area of each leaf was determined using a leaf area meter (AM350 Portable Leaf Area Meter, Hoddesdon, UK); the aboveground fresh weight was determined using an electronic balance; and the aboveground weight of the plants was determined by placing the plants in the oven at 105 °C for 15 min after being stored in the oven and then dried at 80 °C to a constant weight for the determination of their dry weights.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p29
|
PMC11276641
|
sec[3]/sec[2]/sec[1]/p[0]
|
4.3.2. Measurement of Photosynthetic Properties
| 4.082031 |
biomedical
|
Study
|
[
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[
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] |
Measurements of gas exchange were carried out on day 3, day 6, and day 9 of the treatments. Five seedlings were randomly selected from each treatment, and their 2nd true leaves were taken to determine the transpiration rate (E), net photosynthetic rate (Pn), intercellular CO 2 concentration (Ci), and stomatal conductance (gsw) using Plant Photosynthesizer 6800 . The temperature of the leaf chamber was set at 24 °C, the CO 2 level at 400 µmol/mol, and the relative humidity at 60%, and the light source for the determination was set to be R90B10 with a light intensity of 1000 µmol/(m 2 ·s).
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p30
|
PMC11276641
|
sec[3]/sec[2]/sec[2]/p[0]
|
4.3.3. Stomatal Morphology and Electrolyte Permeability (REC)
| 4.125 |
biomedical
|
Study
|
[
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[
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] |
The stomatal pore size was observed on the 9th day of treatment. Three seedlings were randomly selected from each treatment, and the same leaf position of the 2nd leaf was taken to make clinical mounts; the taken leaves were placed on transparent adhesive tape, lightly pressed to make a tight bond, and then lightly scraped with a razor blade to remove the chloroplastic tissues, and the mounts were pasted to the slides. The sections were observed under a microscope (Olympus BX63, Olympus LS, Tokyo, Japan), and stomatal size was measured at 40×. Ten fields of view were selected for each mount, and 20 stomata were selected for each field of view. Pore length and pore width were measured using the ImageJ software (64-bit), and pore area was calculated. Pore area (μm 2 ) = π × pore length/2 × pore width/2.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276641_p31
|
PMC11276641
|
sec[3]/sec[2]/sec[2]/p[1]
|
4.3.3. Stomatal Morphology and Electrolyte Permeability (REC)
| 4.105469 |
biomedical
|
Study
|
[
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] |
The 2nd leaf was selected for each treatment and leaf discs of 1 cm diameter were made using a hole punch in the remaining leaf parts avoiding the main leaf veins. Ten leaf discs were placed in a test tube containing 10 mL of distilled water and immersed in darkness at room temperature for 4 h, during which they were shaken 3–5 times, after which their conductivity was determined as S1 by a conductivity meter, and then the tubes were water-bathed at 100 °C for 20 min, and then shaken after cooling, and then the conductivity was determined as S2. Relative conductivity (%) = (S1 − S0)/(S2 − S0) × 100%, where S0 is the distilled water’s conductivity.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p32
|
PMC11276641
|
sec[3]/sec[2]/sec[3]/p[0]
|
4.3.4. Measurement of Malondialdehyde (MDA) and Superoxide Dismutase (SOD) Enzyme Activity
| 4.09375 |
biomedical
|
Study
|
[
0.99951171875,
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[
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The MDA content was determined by a spectrophotometer using an MDA content assay kit (Solebo, Beijing, China). Weigh 0.1 g of tissue and add 1 mL of extraction solution for ice bath homogenization, centrifuge at 8000× g for 10 min at 4 °C, take the supernatant, and add the reagents sequentially according to the procedure of the manual in a 100 °C water bath for 60 min and then 10,000× g . After centrifugation at room temperature for 10 min, measure the absorbance at 532 nm and 600 nm, and calculate the MDA content according to the manual.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276641_p33
|
PMC11276641
|
sec[3]/sec[2]/sec[3]/p[1]
|
4.3.4. Measurement of Malondialdehyde (MDA) and Superoxide Dismutase (SOD) Enzyme Activity
| 4.082031 |
biomedical
|
Study
|
[
0.99951171875,
0.0002467632293701172,
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] |
[
0.98046875,
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0.0002434253692626953
] |
The content of the SOD activity was determined by a spectrophotometer using a SOD activity assay kit (Solebo, Beijing, China). Weigh 0.1 g of tissue and add 1 mL of extraction solution for ice bath homogenization, centrifuge at 8000× g for 10 min at 4 °C, take the supernatant, add the reagents according to the steps in the instructions in sequence, measure the absorbance at 560 nm after 30 min in a 37 °C water bath, and calculate the SOD activity according to the instructions.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276641_p34
|
PMC11276641
|
sec[3]/sec[2]/sec[4]/p[0]
|
4.3.5. Determination of Hydrogen Peroxide (H 2 O 2 ) and Superoxide Anion (O 2 ∙− ) Content
| 4.054688 |
biomedical
|
Study
|
[
0.99951171875,
0.0002396106719970703,
0.0003325939178466797
] |
[
0.966796875,
0.031951904296875,
0.00079345703125,
0.00029087066650390625
] |
The H 2 O 2 content was determined by a spectrophotometer using an H 2 O 2 content assay kit (Solebo, Beijing, China). Weigh 0.1 g of tissue and add 1 mL of extraction solution for ice bath homogenization, centrifuge at 8000× g at 4 °C for 10 min, take the supernatant, add the reagents sequentially according to the steps in the instruction manual, measure the absorbance at 415 nm, and calculate the H 2 O 2 content according to the instruction manual.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276641_p35
|
PMC11276641
|
sec[3]/sec[2]/sec[4]/p[1]
|
4.3.5. Determination of Hydrogen Peroxide (H 2 O 2 ) and Superoxide Anion (O 2 ∙− ) Content
| 4.050781 |
biomedical
|
Study
|
[
0.99951171875,
0.00022292137145996094,
0.0002994537353515625
] |
[
0.9794921875,
0.0197601318359375,
0.0006818771362304688,
0.00024127960205078125
] |
The O 2 ∙− content was determined by a spectrophotometer using an O 2 ∙− content assay kit (Solebo, Beijing, China). Weigh 0.1 g of tissue and add 1 mL of extraction solution for ice bath homogenization, centrifuge at 12,000 rpm and 4 °C for 20 min, take the supernatant, add the reagents sequentially according to the instruction, measure the absorbance at 530 nm, and calculate the O 2 ∙− content according to the instruction.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276641_p36
|
PMC11276641
|
sec[3]/sec[2]/sec[4]/p[2]
|
4.3.5. Determination of Hydrogen Peroxide (H 2 O 2 ) and Superoxide Anion (O 2 ∙− ) Content
| 2.595703 |
biomedical
|
Study
|
[
0.96923828125,
0.0006856918334960938,
0.0301666259765625
] |
[
0.9560546875,
0.042236328125,
0.0011730194091796875,
0.0003714561462402344
] |
On the 9th day of the treatments carried out, the 2nd true leaves of three melon seedlings from each treatment were selected and sampled for tissue staining. DAB staining and NBT staining were referred to by Hu et al. .
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p37
|
PMC11276641
|
sec[3]/sec[2]/sec[5]/p[0]
|
4.3.6. RNA Extraction and Transcriptome Sequencing
| 4.191406 |
biomedical
|
Study
|
[
0.9990234375,
0.00024044513702392578,
0.0007495880126953125
] |
[
0.9990234375,
0.0005106925964355469,
0.00020253658294677734,
0.00004965066909790039
] |
The 2nd leaf of the melon seedlings under CK and T3 treatments was taken after 9 d of drought treatment, and the total RNA was extracted using Trizol reagent (Thermofisher, Waltham, MA, USA) according to the steps provided by the manufacturer. After extraction of the total RNA, mRNA was purified using Dynabeads Oligo (dT) (Thermofisher, Waltham, MA, USA) from the total RNA (5 µg). The total RNA extracted from each leaf sample was used for RNA-Seq library construction and sequencing by Lianchuan Biotechnology Co. (Beijing, China). After purification, mRNA was fragmented into short fragments using divalent cations at a high temperature using RNA Fragmentation Module at 94 °C for 5–7 min. Then, cDNA was generated by reverse transcription with SuperScript™ II reverse transcriptase , RNase H , and dUTP solution . U-labeled second-strand DNA was synthesized with R0133 The ligated products were amplified by PCR after the treatment of U-labeled second-strand DNA with the thermal UDG enzyme . The average insert fragment size of the final cDNA library was 300 ± 50 bp for the average insert length of the cDNA library. Finally, 2 × 150 bp paired-end sequencing (PE150) was performed on IlluminaNovaseq™6000 (LC-Bio Technology CO., Hangzhou, China).
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276641_p38
|
PMC11276641
|
sec[3]/sec[2]/sec[6]/p[0]
|
4.3.7. Metabolomics Analysis and Differential Metabolite Identification
| 4.226563 |
biomedical
|
Study
|
[
0.99853515625,
0.0003402233123779297,
0.0009732246398925781
] |
[
0.9990234375,
0.0003180503845214844,
0.0004987716674804688,
0.00005644559860229492
] |
The 2nd leaves of melon seedlings under the CK and T3 treatments were taken after 9 days of drought treatment. A total of 100 μL of the sample was taken and mixed with 400 μL of extraction solution (MeOH:ACN, 1:1( v / v )); the extraction solution contained deuterated internal standards, and the mixed solution was vortexed for 30 s, sonicated for 10 min in 4 °C water bath, and incubated for 1 h at 40 °C to precipitate proteins. Then, the samples were centrifuged at 12,000 rpm (RCF = 13,800× g , R = 8.6 cm) for 15 min at 4 °C. The supernatant was transferred to a fresh glass vial for analysis. The quality control (QC) sample was prepared by mixing an equal aliquot of the supernatant of the samples. All samples were collected by an LC-MS system following machine commands. All chromatographic separations were performed using an UltiMate 3000UPLC system (Thermofisher, Waltham, MA, USA) with a reversed-phase separation on an ACQUITY UPLCT3 column (100 mm × 2.1 mm, 1.8 µm, Waters, Milford, MA, USA). The metabolites eluting from the column were detected using a high-resolution tandem mass spectrometer TripleTOF 6600 (SCIEX, Carlsbad, CA, USA). The collected mass spectrometry data were preprocessed using XCMS software (4.7). It was then processed with the XCMS, CAMERA, and metaX toolboxes implemented in the R software (4.2.1). The ions were identified by combining retention time (RT) and m / z data. The metabolites were annotated using the online KEGG and HMDB databases, and the exact molecular mass data ( m / z ) of the samples were matched to the data in the database. Statistical analysis of the data was mainly conducted using the R software (version4.0); a clustering heatmap was plotted by the R package pheatmap; PCA analysis and significantly different protein analyses were conducted using the R package metaX (2.67); PLSDA analysis was performed using the R package ropls, and VIP values of each variable were calculated; correlation analysis was performed using the Pearson correlation coefficient of R package cor, and a p -value < 0.001 was calculated using a T-test obtained by a p -value < 0.05 and a multiplicity with a difference > 1.2; and the VIP calculated using PLSDA analysis was satisfied simultaneously to screen the final significantly different metabolites. Differential enrichment analysis of the KEGG pathway was performed based on the hypergeometric test, and the functional entries with a p -value < 0.05 from the statistical test were the functional entries significantly enriched for differential proteins.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p39
|
PMC11276641
|
sec[3]/sec[2]/sec[7]/p[0]
|
4.3.8. Real-Time Quantitative RT-PCR Assay
| 4.109375 |
biomedical
|
Study
|
[
0.99951171875,
0.0002263784408569336,
0.0003981590270996094
] |
[
0.99951171875,
0.00029468536376953125,
0.00023448467254638672,
0.00004607439041137695
] |
In total, 2 g of leaf tissue was taken into a 2 mL centrifuge tube and quickly frozen in liquid nitrogen, followed by high-frequency shaking using a high-throughput grinder. The reliability of RNA-Seq data was verified by qRT-PCR using nine randomly selected differential genes. Primers were designed by Primer Premier 5.0, and the primer sequences are shown in Table S1 . The total leaf RNA was extracted using the RBA extraction kit (Aidab, Beijing, China) according to the standard step-by-step method of the reagent. A reverse transcription kit (Cofitt, Hangzhou, China) was used. The extracted RNA was reverse transcribed into cDNA using a reverse transcription kit (Cofitt, Hangzhou, China) according to the standard method of the reagent, and the relative expression of the genes was analyzed using the 2 −ΔΔCt analysis method in a 20 μL reaction system according to the 2× qPCR SmArt Mix (SYBR Green) kit (Descartes Biologicals, Shanghai, China).
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276641_p40
|
PMC11276641
|
sec[3]/sec[3]/p[0]
|
4.4. Statistical Analysis
| 1.821289 |
biomedical
|
Study
|
[
0.984375,
0.0019121170043945312,
0.013580322265625
] |
[
0.5888671875,
0.40625,
0.003200531005859375,
0.001819610595703125
] |
Data were collated and plotted using Microsoft Office Excel 2020 and analyzed for significant differences using IBM SPSS Statistics 25.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11276641_p41
|
PMC11276641
|
sec[4]/p[0]
|
5. Conclusions
| 4.148438 |
biomedical
|
Study
|
[
0.99560546875,
0.0004093647003173828,
0.0037593841552734375
] |
[
0.9990234375,
0.00026154518127441406,
0.0004355907440185547,
0.00004309415817260742
] |
On the basis of determining the ratio of red and blue light and photosynthetically active radiation, increasing the ratio of green light instead of red and blue light improved the drought tolerance of melon seedlings; at the same time, it reduced the electrolyte permeability and MDA content of melon seedling leaves and increased O 2 ∙− and H 2 O 2 content, which helped to alleviate the damage of short-term drought to the cell membranes of melon seedlings. The addition of green light initially decreased leaf stomatal conductance and reduced water loss. The transcriptomic and metabolomic results indicated that the process of green light-regulated enhancement of drought tolerance in plants is regulated by multiple phytohormones. This study provides a new perspective for thinking about improving plant water utilization in facilities.
|
[
"Xue Li",
"Shiwen Zhao",
"Qianqian Cao",
"Chun Qiu",
"Yuanyuan Yang",
"Guanzhi Zhang",
"Yongjun Wu",
"Zhenchao Yang"
] |
https://doi.org/10.3390/ijms25147561
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p0
|
PMC11276653
|
sec[0]/p[0]
|
1. Introduction
| 4.257813 |
biomedical
|
Review
|
[
0.99853515625,
0.0009713172912597656,
0.0004200935363769531
] |
[
0.145751953125,
0.049591064453125,
0.8017578125,
0.0027217864990234375
] |
Parkinson’s disease (PD) is a progressive neurodegenerative disorder that is characterized by four major motor symptoms: resting tremor, bradykinesia, rigidity, and loss of postural reflexes . Nonmotor symptoms, such as various psychiatric symptoms and autonomic dysfunction, are also associated with PD, considerably contributing to a decline in quality of life in patients . Neuronal loss in the substantia nigra, which causes striatal dopamine deficiency, and intracellular inclusions that contain aggregates of α-synuclein, are neuropathological hallmarks of PD .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p1
|
PMC11276653
|
sec[0]/p[1]
|
1. Introduction
| 4.222656 |
biomedical
|
Review
|
[
0.99560546875,
0.0023136138916015625,
0.0021610260009765625
] |
[
0.0295562744140625,
0.0014905929565429688,
0.96875,
0.0004115104675292969
] |
Among nonmotor symptoms of PD, apathy consists of a set of behavioral, affective, and cognitive features. Apathy can be defined as a state of low motivation that manifests as a decrease in goal-directed behaviors. It can be variably characterized by a reduction in interests and blunted emotions that cannot be attributed to lower levels of consciousness, cognitive impairment, or emotional distress . The comorbidity of apathy is not limited to advanced stages of PD. It can also be present from early stages of the disease, and its prevalence is reported to range from 16.5% to 60% of PD patients . In PD patients, apathy-associated complications are important and play a key role in daily functioning. For example, apathy can lead to a poor response to motor symptom treatment and greater healthcare spending of both patients and their caregivers, and it can negatively influence general cognitive function and financial capacity performance . Additionally, apathy was reported to be an independent nonmotor symptom that determined poor health-related quality of life in patients with early PD . Although apathy causes such significant losses in the lives of patients, there is no specific evidence-based treatment .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p2
|
PMC11276653
|
sec[0]/p[2]
|
1. Introduction
| 4.195313 |
biomedical
|
Review
|
[
0.99560546875,
0.0007395744323730469,
0.0034942626953125
] |
[
0.3388671875,
0.0048828125,
0.65576171875,
0.0004496574401855469
] |
Various symptoms of apathy can be classified into several subtypes. The responsible neural systems, comprising multiple anatomical sites, for each subtype are assumed to be different and partially interact . Several classifications of apathy have been proposed in the literature, which are generally classified into “cognitive”, “emotional-affective (motivational)”, and “self(auto)-activation”, and there is a tendency to classify symptoms that are related to social interaction separately as “social” apathy . Dysfunction of the mesocorticolimbic pathway leads to emotional-affective apathy, and dysfunction of the dorsolateral prefrontal cortex (PFC) and caudate nucleus leads to cognitive apathy . Although social apathy has not been investigated sufficiently in previous work, the anterior cingulate cortex (ACC) and PFC have been suggested to play a role in processing social information and social interaction . Each apathy subtype exhibits different kinds of symptoms. For example, disruption of the planning aspect of goal-directed behavior leads to cognitive apathy and clinically manifests as lower interest and lower curiosity . Emotional-affective apathy exhibits a blunted emotional response when the patient is confronted with upsetting news or watches something humorous. For example, patients can exhibit little concern for the health and well-being of themselves or their family members . In social apathy, a low level of engagement in social interactions (e.g., patients cannot initiate conversations without being prompted) may be observed .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276653_p3
|
PMC11276653
|
sec[0]/p[3]
|
1. Introduction
| 2.554688 |
biomedical
|
Other
|
[
0.99072265625,
0.0006737709045410156,
0.0086669921875
] |
[
0.208984375,
0.76513671875,
0.0246734619140625,
0.0009050369262695312
] |
Based on these considerations, a single agent is expected to not be effective for treating all symptoms of apathy. Effective treatments may differ depending on symptom subtypes. To develop therapies based on different types of symptoms, correspondence between apathy subtypes and symptoms needs to be investigated in experimental animals.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p4
|
PMC11276653
|
sec[0]/p[4]
|
1. Introduction
| 2.439453 |
biomedical
|
Study
|
[
0.99462890625,
0.0004267692565917969,
0.0047607421875
] |
[
0.65087890625,
0.30859375,
0.0389404296875,
0.0013751983642578125
] |
Experiments in animals are indispensable in the search for effective treatments for symptoms of apathy. Several animal studies have investigated psychiatric symptoms of PD, including apathy . However, few animal studies have focused on behavior from the standpoint of apathy subtypes, including cognitive, emotional-affective, and social aspects of apathy .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p5
|
PMC11276653
|
sec[0]/p[5]
|
1. Introduction
| 4.117188 |
biomedical
|
Study
|
[
0.99951171875,
0.00026535987854003906,
0.00016796588897705078
] |
[
0.99951171875,
0.00021266937255859375,
0.0003333091735839844,
0.00008088350296020508
] |
In the present study, we generated a mouse model of PD with local 6-hydroxydopamine (6-OHDA) administration, which induces dopaminergic neuronal cell death, bilaterally in the dorsal striatum. Based on previous reports , we hypothesized that the corresponding neural network, including the dorsal striatum, would be disrupted and result in behavioral abnormalities that are attributable to the cognitive apathy subtype. We examined which subtype of apathy is actually observed by dopaminergic neuronal loss in the dorsal striatum and the behavioral symptoms that are elicited.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p6
|
PMC11276653
|
sec[0]/p[6]
|
1. Introduction
| 3.074219 |
biomedical
|
Study
|
[
0.9931640625,
0.0002963542938232422,
0.006679534912109375
] |
[
0.9814453125,
0.01428985595703125,
0.00403594970703125,
0.00021851062774658203
] |
Anhedonia was originally conceptualized only as an inability to experience pleasure, but it is now recognized as including the loss of motivation to act in pursuit of pleasure. Although apathy and anhedonia represent different symptoms, recent studies suggest that they partly overlap and share neural mechanisms that are related to reward processing . Therefore, anhedonia was also evaluated in the present study as an apathy-like behavior.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276653_p7
|
PMC11276653
|
sec[0]/p[7]
|
1. Introduction
| 2.078125 |
biomedical
|
Study
|
[
0.99072265625,
0.0019969940185546875,
0.0074310302734375
] |
[
0.89404296875,
0.09906005859375,
0.00524139404296875,
0.001773834228515625
] |
The experimental concept and design are depicted in Figure 1 A,B.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p8
|
PMC11276653
|
sec[1]/sec[0]/p[0]
|
2.1. Decrease in Tyrosine Hydroxylase (TH) Immunostaining in the Caudate Putamen (CPu) and Substantia Nigra Compacta (SNc) by 6-OHDA Injections
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
0.00028014183044433594,
0.00018084049224853516
] |
[
0.99951171875,
0.0002510547637939453,
0.0002429485321044922,
0.00007599592208862305
] |
One mouse in the 6-OHDA group exhibited almost no TH-positive neuronal loss in the SNc, suggesting a failure of 6-OHDA lesioning. Therefore, this mouse was excluded from all evaluations. We analyzed the loss of TH-positive neurons in the CPu and nucleus accumbens (NAcc) as the cephalic slice and in the SNc and ventral tegmental area (VTA) as the caudal slice . Histological analysis revealed the significant loss of TH-positive pixels (mean ± standard error of the mean [SEM]) in the CPu and cell counts in the SNc in the 6-OHDA group compared with the sham group. In the NAcc and VTA, no significant reductions in pixels or cell counts were found in the 6-OHDA group compared with the sham group . These results indicate that TH neurons in the CPu and SNc, but not in the VTA or NAcc, were dominantly lesioned with 6-OHDA in our mouse model.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p9
|
PMC11276653
|
sec[1]/sec[1]/p[0]
|
2.2. Motor Performance Decline in the 6-OHDA Group
| 4.199219 |
biomedical
|
Study
|
[
0.99951171875,
0.0003459453582763672,
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] |
[
0.9990234375,
0.0002028942108154297,
0.00048160552978515625,
0.00007420778274536133
] |
To ensure validity of the mouse model as a model of PD, three motor performance tests (rotarod, pole, and balance beam) were conducted to confirm motor dysfunction. In the rotarod test, the sham group did not change significantly (before sham lesioning: 250.63 ± 19.96 s; after sham lesioning: 259.54 ± 22.58 s; p = 0.84), but the 6-OHDA group exhibited impairments after lesioning . After lesioning, a comparison between the sham group and 6-OHDA group showed a significant decrease in motor performance in the 6-OHDA group . In the pole test, the time required for the mice to orient themselves in a downward direction, T turn , did not indicate impairments in either group before or after lesioning (sham group: before sham lesioning: 2.65 ± 0.32 s, after sham lesioning: 2.29 ± 0.16 s, p = 0.38; 6-OHDA group: before lesioning: 2.61 ± 0.28 s, after lesioning: 3.15 ± 0.39 s, p = 0.47). A comparison between the sham group and 6-OHDA group showed no significant impairments in the 6-OHDA group ( p = 0.062). For the time required for the mice to descend to the base of the pole, T down , in the pole test, neither group changed significantly (sham group: before sham lesioning: 9.26 ± 0.80 s, after sham lesioning: 8.48 ± 0.45 s, p = 0.38; 6-OHDA group: before lesioning: 9.39 ± 0.51 s, after lesioning: 10.28 ± 0.37 s, p = 0.15). After lesioning, a comparison of the sham group and 6-OHDA group showed a significant increase in time in the 6-OHDA group . In the balance beam test, there was no significant change in the sham group (before sham lesioning: 9.33 ± 0.78 s, after sham lesioning: 8.42 ± 0.63 s; p = 0.313). A longitudinal comparison within the 6-OHDA group showed a decline in times, although this was not significant (before lesioning: 8.43 ± 0.31 s; after lesioning: 11.32 ± 1.08 s; p = 0.051). A postoperative comparison of the sham group and 6-OHDA group showed a significant increase in time in the 6-OHDA group . The 6-OHDA group exhibited a decline in most of the motor performance tests, indicating that they could be used as a PD model.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p10
|
PMC11276653
|
sec[1]/sec[2]/p[0]
|
2.3. Anhedonia-like Behavior Suggested by the Novelty-Suppressed Feeding Test (NSFT) but Not by the Sucrose Preference Test (SPT) in the 6-OHDA Group
| 3.837891 |
biomedical
|
Other
|
[
0.99560546875,
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] |
[
0.0909423828125,
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] |
The sucrose preference test (SPT) is a behavioral test that assesses anhedonia, and various protocols exist . Rodents have an innate preference for palatable, sweet solutions. This preference is thought to correlate with the pleasant sensation that is experienced by animals when they ingest sucrose. Therefore, lower sucrose preference is interpreted as a state of lower pleasure or anhedonia . The novelty-suppressed feeding test (NSFT) is used to assess anxiety but is also used to assess a component of anhedonia .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p11
|
PMC11276653
|
sec[1]/sec[2]/p[1]
|
2.3. Anhedonia-like Behavior Suggested by the Novelty-Suppressed Feeding Test (NSFT) but Not by the Sucrose Preference Test (SPT) in the 6-OHDA Group
| 4.070313 |
biomedical
|
Study
|
[
0.99951171875,
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] |
[
0.99951171875,
0.00039839744567871094,
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0.00004965066909790039
] |
In the SPT, the sucrose preference index (percentage) was not different between the sham and 6-OHDA groups . In the NSFT, one mouse in the sham group and three mice in the 6-OHDA group reached the 6 min cutoff time before eating the food pellet. The mean latency to reach the pellet and take a bite with the forelimbs was significantly longer in the 6-OHDA group . The result of the SPT did not suggest anhedonia, but the NSFT did.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276653_p12
|
PMC11276653
|
sec[1]/sec[3]/p[0]
|
2.4. Anxiety Was Not Observed but Novelty-Seeking Behavior Decreased in the 6-OHDA Group
| 4.175781 |
biomedical
|
Study
|
[
0.99951171875,
0.0002123117446899414,
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] |
[
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] |
The open-field test (OFT) can be used as a test of anxiety because as an anxiety-like behavior, mice do not prefer entering the central area of a test space . In the OFT, the number of times each mouse entered the four center square zone was not different between the sham and 6-OHDA groups . The time each mouse was in the four center square zone was also not different . There was no difference in the total distance traveled between the sham and 6-OHDA groups , suggesting that spontaneous locomotor activity was not affected by 6-OHDA. The hole-board test (HBT) is a behavioral test that evaluates novelty-seeking behavior . A decrease in the number of head-dipping behaviors in the novel environment within the time limit suggests a decrease in innate novelty-seeking behavior in rodents. In the HBT, the number of times the mice introduced their head in the holes significantly decreased in the 6-OHDA group compared with the sham group . Furthermore, statistical analysis of the correlation between the HBT results and pathological analysis results in the 6-OHDA group using the Pearson correlation coefficient showed a significant positive correlation with SNc cell counts . These results indicated a significant decrease in novelty-seeking behavior but not anxiety-like behavior.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p13
|
PMC11276653
|
sec[1]/sec[4]/p[0]
|
2.5. Self-Care Behavior Was Maintained in the 6-OHDA Group
| 4.125 |
biomedical
|
Study
|
[
0.9990234375,
0.00030231475830078125,
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] |
[
0.99951171875,
0.0001475811004638672,
0.00025916099548339844,
0.00004887580871582031
] |
As a component of apathy-like symptoms, self-care activity was assessed in the nest-building test (NBT) and splash test (ST). The NBT can be used to examine apathy symptoms , although a wide variety of causes, such as general condition, distress, and pain, may also provoke deficits in nesting behavior . The ST is used to examine apathy symptoms and self-care behavior . In the NBT, the nest-building score decreased in the 6-OHDA group (1.45 ± 0.17) compared with the sham group (2.13 ± 0.30), but this difference was not statistically significant . In the ST, the latency to the first grooming bout was longer in the 6-OHDA group (6.23 ± 0.37 s) compared with the sham group (2.99 ± 1.48 s), but this difference was also not statistically significant ( p = 0.074), and the SDs were large. The total number of grooming behaviors was also not significantly different between the sham and 6-OHDA groups . In the NBT, a nonsignificant tendency toward a decrease in nesting activity was observed. No apparent decline in grooming behavior was indicated in the ST. In summary, these results did not suggest an obvious impairment in self-care behavior.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p14
|
PMC11276653
|
sec[1]/sec[5]/p[0]
|
2.6. Object/Inanimate Novelty Was Impaired but Social Novelty Was Maintained in the 6-OHDA Group
| 4.109375 |
biomedical
|
Study
|
[
0.99951171875,
0.0003094673156738281,
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] |
[
0.99951171875,
0.00018584728240966797,
0.0003027915954589844,
0.00005453824996948242
] |
As social interaction was assessed in the three-chamber test. The first session (session 1) assessed preference for a social object (mouse) compared with a novel, nonanimal object. Another session (session 2) assessed preference for a newer individual rather than a familiar one . The time that the head of the mouse was within the zone 30 mm around the holding cage (interaction time) was assessed. In session 1, assessment of sociability, the interaction time for the novel object significantly decreased in the 6-OHDA group compared with the sham group . There was no significant difference in interaction time for the social object between the two groups . These results indicate that social behavior was maintained in both groups of mice. However, the exploration time for a novel object significantly decreased in the 6-OHDA group. In session 2, the social novelty preference test, the interaction time for the familiar mouse and stranger mouse did not significantly differ between the two groups . These results indicate that social novelty seeking was maintained.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p15
|
PMC11276653
|
sec[2]/p[0]
|
3. Discussion
| 4.101563 |
biomedical
|
Study
|
[
0.99951171875,
0.000278472900390625,
0.00021183490753173828
] |
[
0.99951171875,
0.00012803077697753906,
0.0004494190216064453,
0.00006628036499023438
] |
In the present study, we adopted 6-OHDA lesioning methods that have been used extensively in previous studies as a mouse model of PD, including studies of nonmotor symptoms . Although negative opinions have begun to be advocated in recent years, male mice have been utilized to avoid confounding effects of the female estrous cycle and other potential variables that may interfere with experimental outcomes . Following this conventional practice, we used only male mice in the present experiments. The validity of the PD model was confirmed by motor tests and pathological evaluations. Behavioral tests of apathy-like behavior were then conducted. Significant differences were found in the HBT and three-chamber test, suggesting a reduction in object/inanimate novelty seeking in the 6-OHDA group. Significant differences were also found in the NSFT. The present study is novel and valuable in that animal behavioral experiments showed that reduced novelty-seeking behavior seems to be a symptom with a strong cognitive apathy component among the apathy symptoms of PD. Furthermore, it is interesting to note that while object/inanimate novelty seeking decreased, social novelty was maintained.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p16
|
PMC11276653
|
sec[2]/p[1]
|
3. Discussion
| 4.101563 |
biomedical
|
Study
|
[
0.99951171875,
0.00019073486328125,
0.00017774105072021484
] |
[
0.99951171875,
0.00020694732666015625,
0.000347137451171875,
0.000064849853515625
] |
The administration of 6-OHDA reduced dopaminergic neurons in the CPu and SNc to 69% and 68% of the sham group, respectively. The percentage of dopamine neuronal loss in the striatum was reported to be 63–74% in a previous study . The degree of dopaminergic neuronal loss in the present study was at the same level and is considered to correspond to the early stage of PD. Motor performance significantly decreased in our model, indicating that the PD mouse model may mimic an earlier stage of PD. In the pole test, T turn measures the time required for the mouse to change head direction, and T down measures the time to descend the pole. The present results showed significant differences only in T down and not in T turn , which may indicate mild motor symptoms in the present mouse model.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p17
|
PMC11276653
|
sec[2]/p[2]
|
3. Discussion
| 4.09375 |
biomedical
|
Study
|
[
0.99951171875,
0.00014853477478027344,
0.00015175342559814453
] |
[
0.9990234375,
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0.0005164146423339844,
0.00007551908493041992
] |
Apathy is reported to occur at any stage of PD in human patients, even in the prodromal stage when motor symptoms are not evident . Based on the results of the motor function tests and pathological evaluation, our model mice correspond to the early stage of PD. Notably, 6-OHDA-lesioned mice exhibited motor symptoms despite a lesser degree of dopaminergic degeneration compared with human PD patients . This may be because 6-OHDA administration induced rapid dopaminergic denervation in animals, whereas this process takes several years to decades in humans.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p18
|
PMC11276653
|
sec[2]/p[3]
|
3. Discussion
| 4.339844 |
biomedical
|
Study
|
[
0.99951171875,
0.00033593177795410156,
0.000263214111328125
] |
[
0.9990234375,
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] |
In the experiments of apathy-like behavior, we found that the 6-OHDA group exhibited significant impairments in novelty seeking. Notably, PD patients may exhibit low novelty-seeking behavior . Although the underlying neural mechanism of novelty-seeking behavior in PD patients is largely unknown, it is assumed that the midbrain dopaminergic system plays an essential role . The activity of VTA/SNc dopaminergic neurons has been reported to correlate with an increase or a decrease in novelty-seeking behavior in mice . The present PD mouse model exhibited dopaminergic neuronal loss in the dorsal striatum and impairments in novelty seeking. Low novelty seeking, which was observed in the present study, is considered a symptom with a strong cognitive apathy component in PD. The dorsal striatum, which was lesioned in the present study, is thought to be involved in cognitive apathy . Additionally, a detailed anatomical study found that the dorsolateral PFC is involved in executive processing in cooperation with the ACC in cognitive apathy. The dorsolateral PFC projects neural circuits to the dorsal striatum , which was the target of 6-OHDA lesioning in our PD model. Therefore, our mice may be considered a model of cognitive apathy, especially low novelty seeking. With regard to novelty seeking, recent meta-analyses revealed that in personality profiles, PD patients tend to exhibit significantly lower tendencies to explore novelty compared with healthy controls . These changes in personality and motivational tendencies are known as hypodopaminergic behavioral patterns . Dopamine receptor agonist administration in PD patients who performed a feedback-based probabilistic classification task showed an increase in novelty seeking . Thus, lower novelty seeking may be a more universal problem among PD patients in general, with or without concomitant apathy. Few studies have examined novel exploratory behavior in animal models of PD . Thus, the present results provide important insights into this universal problem among PD patients.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276653_p19
|
PMC11276653
|
sec[2]/p[4]
|
3. Discussion
| 3.806641 |
biomedical
|
Study
|
[
0.9990234375,
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] |
[
0.9990234375,
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0.000056862831115722656
] |
The latency to eat food was significantly longer in the 6-OHDA group in the NSFT, suggesting that these mice exhibited anhedonia. In contrast, no difference in the percentage of sucrose solution consumption was found between the sham group and 6-OHDA group in the SPT, another test of anhedonia.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276653_p20
|
PMC11276653
|
sec[2]/p[5]
|
3. Discussion
| 4.078125 |
biomedical
|
Study
|
[
0.9990234375,
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] |
[
0.99951171875,
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] |
In the SPT, the score dispersion was particularly high in the sham group. This may be attributable to bias that is caused by some factors. With regard to the test duration, one rat study suggested that 1 h is insufficient because the animals drink water regardless of taste immediately after 24 h of water deprivation because of thirst . Hence, we conducted a 2 h SPT, although this may have not been sufficient and led to the score dispersion. Furthermore, we imposed water deprivation according to previous studies that reported that 10–15% of mice do not drink water without first being water deprived ; thus, we imposed water deprivation to avoid this caveat. Nevertheless, water deprivation was shown to lead to an increase in the preference threshold for the sucrose solution and may affect the rate of development of preference and thus the overall results .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p21
|
PMC11276653
|
sec[2]/p[6]
|
3. Discussion
| 2.587891 |
biomedical
|
Study
|
[
0.9775390625,
0.0004799365997314453,
0.0217742919921875
] |
[
0.98876953125,
0.0106201171875,
0.0003685951232910156,
0.00014662742614746094
] |
A motivational factor was added in the NSFT. In the NSFT, the desire to eat under fasting conditions conflicts with the fear of novel spaces . Although the fear of novel spaces has some similarities to the OFT, no anxious behavior was suggested in the present study. However, the reduction in exploratory behavior, as suggested by the HBT results, may have influenced the NSFT in the 6-OHDA group.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276653_p22
|
PMC11276653
|
sec[2]/p[7]
|
3. Discussion
| 3.953125 |
biomedical
|
Study
|
[
0.9990234375,
0.00018322467803955078,
0.0007853507995605469
] |
[
0.9990234375,
0.0004036426544189453,
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0.00004887580871582031
] |
Additionally, motor impairments (i.e., the latency of action) may more greatly affect performance in the NSFT than in the ST. However, the total distance traveled and latency from the start of the test to transfer to another zonein the OFT did not significantly differ between the 6-OHDA and sham groups. This suggests that the motor effect was not considerable. However, we did not assess these parameters in the NSFT. Another possibility is that the conflicting results between the NSFT and SPT might be explained by lower motivation to eat or a decline in the awareness of one’s hunger state, but further studies are required to investigate this possibility.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p23
|
PMC11276653
|
sec[2]/p[8]
|
3. Discussion
| 4.0625 |
biomedical
|
Study
|
[
0.9990234375,
0.00017905235290527344,
0.0007152557373046875
] |
[
0.99951171875,
0.00024962425231933594,
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0.00004571676254272461
] |
Interestingly, a decrease in object/inanimate novelty seeking was suggested by the HBT results and session 1 in the three-chamber test, but social novelty seeking was maintained in the 6-OHDA group in session 2 in the three-chamber test. The possibility that VTA/SNc dopaminergic neurons also mediate social novelty has been proposed . However, a recent study suggests that another midbrain region, the interpeduncular nucleus, but not the VTA, controls social novelty familiarization . These findings and the present results suggest that different neural mechanisms beyond the dorsal striatum may play important roles in social novelty seeking.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p24
|
PMC11276653
|
sec[2]/p[9]
|
3. Discussion
| 4.082031 |
biomedical
|
Study
|
[
0.99951171875,
0.00014853477478027344,
0.00031685829162597656
] |
[
0.9990234375,
0.0004608631134033203,
0.00028228759765625,
0.00006008148193359375
] |
As described above, the present mouse model of PD exhibited cognitive apathy-like behavior. However, emotional-affective apathy was not observed. Indeed, emotional-affective apathy is mediated by the mesocorticolimbic pathway or the reward system. The involved regions include the orbitomedial PFC, ACC, ventral striatum, ventral pallidum, and dopaminergic midbrain neurons. This system is mediated by the amygdala, hippocampus, thalamus, lateral habenular nucleus, dorsal PFC, pedunculopontine nucleus, and raphe nucleus in the brainstem . In the present study, the lesion was predominantly located in the dorsal striatum and there is little direct involvement with the foci responsible for the emotional-affective apathy proposed above. Therefore, this type of apathy is less likely to be related to our model mouse.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p25
|
PMC11276653
|
sec[2]/p[10]
|
3. Discussion
| 4.109375 |
biomedical
|
Study
|
[
0.99951171875,
0.00027370452880859375,
0.00025010108947753906
] |
[
0.99951171875,
0.00013184547424316406,
0.0003612041473388672,
0.00005817413330078125
] |
The present study has several limitations. One limitation of the present study was that cholinergic neurons were not manipulated or assessed in the present mouse model. Cholinergic neurons are also engaged in the nervous system to impact apathy, especially the cognitive subtype . Although studies of the relationship between neuropsychiatric symptoms of PD and cholinergic dysfunction are limited, an interplay between cholinergic dysfunction and neuropsychiatric symptoms has been suggested . On the other hand, some previous studies reported the compensatory upregulation of acetylcholine in PD patients . Abnormalities in dopamine neurons are also associated with executive dysfunction or cognitive dysfunction . Therefore, we presume that dopamine loss itself may cause symptoms of apathy, regardless of changes in cholinergic function. Future studies will reveal whether the cholinergic system is involved in the appearance of apathy-like symptoms in these mice. Next, we describe limitations on the experimental method. Although the number of mice that were tested in the present study may be relatively small, recent studies of psychiatric symptoms that used the 6-OHDA lesioning model of PD in mice had group sizes of 6 to 11, reporting significant differences in the behavioral results . When possible, it is preferable to reduce the number of animals used from an animal ethics standpoint. Even if we used desipramine, we cannot totally exclude possible toxic effects on noradrenergic neurons, which would affect PD symptoms . Anxiety-like behavior was not suggested by the OFT in the present study. Other tests, such as the elevated plus maze test, would have been a good additional control for the absence of anxiety-like behavior.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p26
|
PMC11276653
|
sec[3]/sec[0]/p[0]
|
4.1. Animals
| 2.898438 |
biomedical
|
Study
|
[
0.99853515625,
0.0006995201110839844,
0.0008502006530761719
] |
[
0.8076171875,
0.189208984375,
0.0016727447509765625,
0.0015439987182617188
] |
Eight-week-old male C57BL/6J mice (21–25 g; CLEA Japan, Tokyo, Japan) were housed in our animal facility that was maintained at 23 °C ± 1 °C and 55% ± 5% relative humidity under a 12 h/12 h light/dark cycle (lights on at 8:00 AM, off at 8:00 PM). Food and water were freely available. The experimental procedures and housing conditions were approved by the Institutional Animal Care and Use Committee (Animal Experimentation Ethics Committee, Tokyo Metropolitan Institute of Medical Science; approval no. 23-009). All animals were cared for and treated humanely in accordance with our institutional animal experimentation guidelines.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p27
|
PMC11276653
|
sec[3]/sec[1]/p[0]
|
4.2. Experimental Design
| 4.0625 |
biomedical
|
Study
|
[
0.99951171875,
0.0002856254577636719,
0.00025153160095214844
] |
[
0.99951171875,
0.00020253658294677734,
0.00021779537200927734,
0.000057697296142578125
] |
The experiments began after 2 weeks of habituation (days 1–14) to our animal facility. Three motor performance tests, including the rotarod, pole, and balance beam tests, were conducted before 6-OHDA lesioning (pretest). A total of 21 mice were assigned to sham (n = 8) and 6-OHDA lesion (n = 13) groups to ensure that there were no differences in the motor performance tests. The 6-OHDA lesioning procedure was then performed. After 3 weeks of recovery from lesioning, when the effect of 6-OHDA was expected to stabilize , motor performance tests were conducted again (posttest), and then seven tests that are related to apathy-like behavior were conducted. To avoid influencing each other, the behavioral experiments were conducted during different periods of time, with intervals of a few days between them, based on previous reports . We then performed TH immunostaining. All raw data are provided in Supplementary Materials .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p28
|
PMC11276653
|
sec[3]/sec[2]/p[0]
|
4.3. 6-Hydroxydopamine Lesioning (Day 29)
| 4.175781 |
biomedical
|
Study
|
[
0.99853515625,
0.001377105712890625,
0.00019538402557373047
] |
[
0.99853515625,
0.0010929107666015625,
0.0003104209899902344,
0.00021374225616455078
] |
All mice were intraperitoneally (i.p.) anesthetized with a mixture of Domitor (1.0 mg/mL; Nippon Zenyaku Kogyo, Fukushima, Japan), midazolam (5 mg/mL; Astellas Pharma, Tokyo, Japan), butorphanol (5.0 mg/mL; Meiji Seika Pharma, Tokyo, Japan), and water and mounted in a stereotaxic frame (Muromachi Kikai, Tokyo, Japan). All mice were administered 25 mg/kg desipramine (Sigma-Aldrich, St. Louis, MO, USA) that was dissolved in saline 30 min before the injection of 6-OHDA (Sigma-Aldrich) to prevent lesions of noradrenergic fibers. 6-OHDA was dissolved in saline that contained 0.02% ascorbic acid. The 6-OHDA group mice received bilateral injections of 1 μL of 6-OHDA (4 μg/μL) in the dorsolateral striatum according to the following coordinates relative to the bregma, with reference to a previous report : anterior/posterior, +0.5 mm; medial/lateral, ±2.2 mm; dorsal/ventral, −3.0 mm. Control sham-lesioned mice were injected with the same volume of vehicle (0.9% saline and 0.02% ascorbic acid). After surgery, the animals were allowed to recover for 3 weeks. Immediately and the next day after surgery, all mice received fluid therapy (5% sterile glucose solution, i.p., 10 mL/kg) to prevent dehydration. Among all individual mice, one mouse from the 6-OHDA group died 13 days after lesioning. The postoperative survival rate in the 6-OHDA group was 92.3%.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p29
|
PMC11276653
|
sec[3]/sec[3]/sec[0]/p[0]
|
4.4.1. Accelerating Rotarod (Days 15 and 16 for the Pretest and Days 50 and 51 for the Posttest)
| 4.070313 |
biomedical
|
Study
|
[
0.99951171875,
0.0002536773681640625,
0.00021064281463623047
] |
[
0.9990234375,
0.0007686614990234375,
0.0003788471221923828,
0.0000680685043334961
] |
The accelerating rotarod test was performed using a rotarod apparatus (MK630B, Muromachi Kikai). The apparatus consisted of a computer-controlled motor-driven single rotating rod (30 mm diameter). The rotation speed of the rod accelerated from 4 rotations per minute (rpm) to 40 rpm over the next 5 min. Mice that fell from the rotating rod were detected automatically by a pressure plate at the base of the apparatus. On day 1 of the test (training), the mice were placed on the rotarod for three trials at 15 min intervals. On day 2 (test), the mice were placed on the rotarod for three trials as described above, and the average latency to fall from the rotating rod was recorded for each individual .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p30
|
PMC11276653
|
sec[3]/sec[3]/sec[1]/p[0]
|
4.4.2. Pole Test (Days 18 and 19 for the Pretest and Days 53 and 54 for the Posttest)
| 4.074219 |
biomedical
|
Study
|
[
0.99951171875,
0.0002505779266357422,
0.00031566619873046875
] |
[
0.9990234375,
0.0007085800170898438,
0.00028228759765625,
0.000056684017181396484
] |
A pole test apparatus, consisting of a wooden vertical pole (8 mm diameter, 600 mm height) with a small disk at the top to prevent mice from climbing over the pole, was placed in the mouse’s home cage with soft bedding. The mice were placed individually on top of the pole with their head oriented upward and allowed to descend to the floor. On day 1 (training), the mice underwent five trials to learn how to descend the pole. On day 2 (test), five pole descent trials were performed, and the average time required for the mice to orient themselves in a downward direction (T turn ) and descend to the base of the pole (T down ) were measured . If the mice slipped down the pole or stopped during a trial, then the results were excluded from the analysis.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p31
|
PMC11276653
|
sec[3]/sec[3]/sec[2]/p[0]
|
4.4.3. Balance Beam Test (Days 22–24 for the Pretest and Days 57–59 for the Posttest)
| 4.070313 |
biomedical
|
Study
|
[
0.99951171875,
0.0002884864807128906,
0.000377655029296875
] |
[
0.9990234375,
0.0005774497985839844,
0.0002294778823852539,
0.000058710575103759766
] |
A custom-built wooden bar (1 m length, 6 mm width) was placed 500 mm above a desk, with one end placed in a dark escape home cage. Testing was performed across 3 consecutive days. During the first 2 days (training), the mice were habituated to the escape cage for 2 min, then placed at the starting point individually, and trained to cross the balance beam to reach the escape cage three times. On the third day (test), the mice were placed at the starting point and allowed to cross the balance beam. Each mouse was tested three times, and the average time to cross the beam was measured . If a mouse stopped in the middle of the bar for more than 2 s, then the results were excluded from the analysis.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p32
|
PMC11276653
|
sec[3]/sec[4]/sec[0]/p[0]
|
4.5.1. Sucrose Preference Test (Days 79–81)
| 4.058594 |
biomedical
|
Study
|
[
0.99951171875,
0.00023186206817626953,
0.00022423267364501953
] |
[
0.99951171875,
0.00031375885009765625,
0.0003154277801513672,
0.00005692243576049805
] |
Anhedonia-related behavior was measured using the SPT . After an 18 h period of water deprivation, each mouse was exposed to two bottles, one containing tap water and the other containing 1% sucrose solution. During the 2 h experimental time (9:00–11:00 AM), each mouse could drink freely from two bottles. The positions of the two bottles were exchanged at 10:00 AM (1 h after the start of the experiment). We conducted a control experiment to check for water leakage from the bottles and confirmed that leakage was negligibly small. Water consumption was measured by weighing the bottle before and after the test. Sucrose preference (expressed as a percentage) was calculated according to the following formula: sucrose solution intake/(sucrose solution + tap water intake).
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p33
|
PMC11276653
|
sec[3]/sec[4]/sec[1]/p[0]
|
4.5.2. Novelty-Suppressed Feeding Test (Days 64 and 65)
| 3.875 |
biomedical
|
Study
|
[
0.99951171875,
0.00020253658294677734,
0.0004475116729736328
] |
[
0.998046875,
0.0016107559204101562,
0.0003275871276855469,
0.00006645917892456055
] |
The mice were food deprived 18 h before the NSFT. The mice were placed in the same corner of a novel, brightly lit plastic cage (290 mm × 460 mm × 260 mm) that all tested mouse had never experienced before the test. A single food pellet that was placed in the center for a 6 min test, and the latency to take a bite of the pellet while holding it with the forelimbs was recorded .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p34
|
PMC11276653
|
sec[3]/sec[4]/sec[2]/p[0]
|
4.5.3. Open-Field Test (Day 86)
| 4.03125 |
biomedical
|
Study
|
[
0.99951171875,
0.00022685527801513672,
0.0002732276916503906
] |
[
0.9990234375,
0.0005283355712890625,
0.0002713203430175781,
0.000057578086853027344
] |
The mice were placed individually in the center of a box that was constructed with a white acrylic wall (400 mm × 400 mm × 300 mm) at the bottom. The activity of the mice was observed for 5 min and automatically recorded and analyzed using animal behavior analysis software (ANY-MAZE version 6.1, Stoelting, Wood Dale, IL, USA). In the analysis, the area was divided into 16 equal squares. The number of times each mouse entered the four center square zone (25% of total area), the time that the mice were in the four center square zone, and the total distance traveled in the test were calculated .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p35
|
PMC11276653
|
sec[3]/sec[4]/sec[3]/p[0]
|
4.5.4. Hole-Board Test (Day 89)
| 3.660156 |
biomedical
|
Study
|
[
0.9990234375,
0.0002409219741821289,
0.0009469985961914062
] |
[
0.9892578125,
0.0100250244140625,
0.00044918060302734375,
0.00014960765838623047
] |
The HBT was performed with an HBT apparatus (ST-1/WII, Muromachi Kikai). The apparatus consisted of a gray vinyl chloride box (400 mm × 400 mm × 300 mm) that had four holes in the floor. Each mouse was placed inside the box, and the number of times the mouse introduced its head in these holes was automatically counted by infrared sensors. The test lasted 3 min .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p36
|
PMC11276653
|
sec[3]/sec[4]/sec[4]/p[0]
|
4.5.5. Nest-Building Test (Day 71)
| 4.105469 |
biomedical
|
Study
|
[
0.99951171875,
0.000324249267578125,
0.00040030479431152344
] |
[
0.9990234375,
0.0004982948303222656,
0.0002703666687011719,
0.000056803226470947266
] |
We conducted the NBT according to a previously reported protocol . Each mouse was placed in a new cage in which the entire floor was covered with a Palsoft sheet (i.e., a sheet-type bedding material; Oriental Yeast, Tokyo, Japan) for 5 h (10:00 AM–3:00 PM). At the end of the test, each cage was photographed from two angles: top and side. Nest-building activity was assessed by visual examination of photographs of the formation of a nest according to a previously described scoring method (0, no sign of interaction or manipulation of the bedding material; 1, interaction with the nesting material is evident, but the material has not been gathered to a nest site; 2, nesting material has been gathered to form a flat nest with incomplete walls; 3, nesting material has been gathered to form a nest site and which has identifiable walls that form a “cup” or “bowl” shape; 4, bedding material has been gathered to form a nest site in the cage, and its walls do not constitute an incomplete dome structure; 5, nesting material has been gathered to form a nest site in the cage, and its walls completely enclose a hollow nest) . In addition to this six-point (0–5) scale, 0.5 was added to the integers for intermediate cases.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p37
|
PMC11276653
|
sec[3]/sec[4]/sec[5]/p[0]
|
4.5.6. Splash Test (Day 68)
| 3.697266 |
biomedical
|
Study
|
[
0.9990234375,
0.000431060791015625,
0.000583648681640625
] |
[
0.9951171875,
0.004444122314453125,
0.00048661231994628906,
0.00014865398406982422
] |
The ST started by spraying a 10% sucrose solution twice on the dorsal coat of the mouse. Because of the high viscosity of the sucrose solution, it dirties the mouse’s fur, which induces grooming behavior. After spraying the sucrose solution, behavior was videotaped for 5 min. The latency to the first bout of grooming behavior and total number of grooming bouts were assessed .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p38
|
PMC11276653
|
sec[3]/sec[4]/sec[6]/p[0]
|
4.5.7. Three-Chamber TEST (Days 73–75)
| 3.925781 |
biomedical
|
Study
|
[
0.9990234375,
0.00019931793212890625,
0.0006299018859863281
] |
[
0.99853515625,
0.0013885498046875,
0.0002460479736328125,
0.00005799531936645508
] |
The three-chamber test was conducted using a sociability apparatus (SC-03M, Muromachi Kikai) to assess sociability and social novelty preference . The apparatus consisted of three chambers. The dividing walls of the chamber were constructed of transparent plastic with small square openings (50 mm × 30 mm) that allowed access to each chamber (200 mm × 400 mm × 220 mm). The test mice were first placed in the middle chamber and allowed to explore the entire test chamber for 10 min (habituation).
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276653_p39
|
PMC11276653
|
sec[3]/sec[4]/sec[6]/p[1]
|
4.5.7. Three-Chamber TEST (Days 73–75)
| 4.097656 |
biomedical
|
Study
|
[
0.99951171875,
0.0002963542938232422,
0.0002627372741699219
] |
[
0.99951171875,
0.0003418922424316406,
0.0002884864807128906,
0.00005733966827392578
] |
After that habituation, the three-chamber test consisting of two sessions of 10 min each was conducted. Immediately after the 10 min period, the test mice were placed in a clean holding cage. A small round wire cage (90 mm diameter, 185 mm height, with vertical bars 7 mm apart) was located in the left and right chambers, and a 25 mm sponge cube (novel object) or a 14-week-old male C57BL/6J mouse with no prior contact with the test mice was enclosed in each of the wire cages. Next, the test mice were returned to the middle chamber and allowed to explore for 10 min (sociability test; session 1). After the session, the 14-week-old male C57BL/6J mouse was again placed in the holding cage (familiar), and a second unfamiliar mouse (stranger) was enclosed in the other wire cage on the opposite side. The test mice were placed in the middle chamber and had a choice between the first, already investigated familiar mouse and the novel unfamiliar mouse for 10 min (social novelty preference test; session 2). These experiments were automatically recorded and analyzed using animal behavior analysis software (ANY-MAZE). In the analysis, the interaction zone was set at 30 mm around the holding cage. As an interaction time, the total time that the head of the test mouse was within the zone was counted.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p40
|
PMC11276653
|
sec[3]/sec[5]/p[0]
|
4.6. Immunohistochemistry
| 4.21875 |
biomedical
|
Study
|
[
0.99951171875,
0.0004229545593261719,
0.00016307830810546875
] |
[
0.998046875,
0.0011615753173828125,
0.0005078315734863281,
0.00013768672943115234
] |
After the behavioral tests, the mice were deeply anesthetized with pentobarbital, and then the transcardiac perfusion with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde (PFA) was conducted, using a perfusion pressure pump. The PFA-perfusioned brain was removed and immersed in 4% PFA overnight. The solution was replaced with PBS and stored at 4 °C. Paraffin-embedded tissue sections (5 μm thick) were cut with a sliding microtome. For the pathological evaluation, two different levels of slices were configured according to a mouse brain atlas : (i) CPu and NAcc as the cranial level (bregma +0.86 mm) and (ii) SNc and VTA as the caudal level (bregma −3.08 mm). Immunohistochemistry was conducted as described previously . Briefly, the paraffin-embedded sections were deparaffinized, rehydrated, and immersed in distilled water. The sections were then autoclaved in 0.01 M citrate buffer (pH 6.0) for antigen activation. The sections were immersed in 0.3% hydrogen peroxide to remove endogenous peroxidase and treated with 5% normal goat serum for blocking. The sections were incubated with mouse anti-TH diluted in PBS with 1% fetal bovine serum. Incubations with primary antibodies were performed overnight at 4 °C. The sections were then incubated for 1 h at room temperature with biotinylated secondary antibody (goat anti-mouse immunoglobulin G, 1:200; Vector Laboratories, Newark, CA, USA) diluted in PBS. Afterward, the sections were incubated with avidin peroxidase (1:100 in PBS) for 50 min at room temperature. The reaction product was visualized using diaminobenzidine (Sigma) in TBS with 0.0024% H 2 O 2 .
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276653_p41
|
PMC11276653
|
sec[3]/sec[6]/p[0]
|
4.7. Quantitative Analysis of TH-Positive Cells
| 4.140625 |
biomedical
|
Study
|
[
0.99951171875,
0.00026988983154296875,
0.00021350383758544922
] |
[
0.99951171875,
0.0003306865692138672,
0.00025463104248046875,
0.00006091594696044922
] |
The immunostained slides were photographed using an inverted fluorescence phase-contrast microscope (BZX800, Keyence, Osaka, Japan). For assessment of the CPu and NAcc, bregma +0.86 mm levels were taken as the cephalic slice. For assessment of the SNc and VTA, bregma −3.08 mm levels were taken as the caudal slice. In each slice, a mouse brain atlas was used as a reference for the identification of areas to be evaluated . All of the following steps were performed using ImageJ version 1.53e (National Institutes of Health, Bethesda, MD, USA). For the cephalic slice images, the photographs were converted to 16-bit monochrome images. Background regions were defined in slices that did not contain dopaminergic nerves (lateral and medial septal nucleus). The mean + 2 standard deviations of the contrast in the background region was used as the threshold, and the number of pixels in the target region that showed a contrast higher than the threshold was counted as positive for TH staining. For the caudal slice images, we used a cell count method for TH-positive cells. In the 6-OHDA group, a mouse with TH+ cell counts in the SNc that were >80% of the average of the Sham group was excluded from the analysis because of insufficient lesioning.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276653_p42
|
PMC11276653
|
sec[3]/sec[7]/p[0]
|
4.8. Statistical Analysis
| 3.941406 |
biomedical
|
Study
|
[
0.99951171875,
0.00016760826110839844,
0.00025153160095214844
] |
[
0.99853515625,
0.0008711814880371094,
0.0005135536193847656,
0.00006365776062011719
] |
The statistical analysis was performed using R version 4.0.5 software (R Core Team ; R Foundation for Statistical Computing, Vienna, Austria). The results are presented as the mean ± standard error of the mean (SEM). The assumption of normality was tested for all continuous variables by evaluating the frequency distribution histogram and conducting the Shapiro–Wilk test. For comparisons of means between the sham and 6-OHDA groups, normally distributed data were analyzed using Student’s t -test or Welch’s t -test. The Mann–Whitney U test was conducted for non-normally distributed variables. For comparisons of results over time within the same group, normally distributed data were analyzed using a paired t -test and Wilcoxon signed-rank sum test for non-normally distributed variables. All of the tests were two-tailed. Values of p < 0.05 were considered statistically significant.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276653_p43
|
PMC11276653
|
sec[4]/p[0]
|
5. Conclusions
| 4.097656 |
biomedical
|
Study
|
[
0.99951171875,
0.0001742839813232422,
0.000152587890625
] |
[
0.9990234375,
0.00048470497131347656,
0.0006284713745117188,
0.00008946657180786133
] |
The present mouse model of PD that was induced by bilateral 6-OHDA injections in the dorsal striatum predominantly exhibited a decrease in novelty seeking as a symptom that is related to the cognitive apathy component of PD. Although object/inanimate novelty seeking decreased, social novelty seeking was maintained, suggesting the involvement of different neural circuits beyond the dorsal striatum. The present mouse model of PD may contribute to future therapeutic investigations of apathy based on its subdomains, especially in the treatment of cognitive-like apathy symptoms, including impairments in novelty seeking.
|
[
"Masato Okitsu",
"Masayo Fujita",
"Yuki Moriya",
"Hiroko Kotajima-Murakami",
"Soichiro Ide",
"Rika Kojima",
"Kazunari Sekiyama",
"Kazushi Takahashi",
"Kazutaka Ikeda"
] |
https://doi.org/10.3390/ijms25147993
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p0
|
PMC11276665
|
sec[0]/p[0]
|
1. Introduction
| 4.058594 |
biomedical
|
Review
|
[
0.9931640625,
0.0043792724609375,
0.0025997161865234375
] |
[
0.02105712890625,
0.123291015625,
0.85400390625,
0.0017271041870117188
] |
Pregnancy is characterized by a cascade of physiological changes and adaptations aimed at supporting the growth and development of the fetus . From conception to birth, the pregnant woman navigates a complex interplay of hormonal, metabolic, and immunological shifts, all orchestrated to nurture and sustain the developing life within. During this time, the maternal body provides the necessary nutrients, oxygen, and protection for the growing fetus to thrive. However, the marvel of pregnancy comes with inherent risks and challenges, highlighting the critical nature of this period . Complications arising during pregnancy can have far-reaching implications, both impacting immediate health outcomes and predisposing individuals to a spectrum of chronic diseases later in life . Therefore, ensuring maternal health during pregnancy is essential for the successful progression of gestation and for laying the foundation for the long-term well-being of both the mother and child.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11276665_p1
|
PMC11276665
|
sec[0]/p[1]
|
1. Introduction
| 4.238281 |
biomedical
|
Study
|
[
0.99951171875,
0.00020301342010498047,
0.00016438961029052734
] |
[
0.9912109375,
0.0032482147216796875,
0.0055389404296875,
0.00020563602447509766
] |
The placenta is critical to a healthy pregnancy outcome since it takes on the multifaceted role of supporting fetal growth and ensuring the well-being of both the developing fetus and the mother . Moreover, it also acts as an endocrine organ by synthetizing and secreting various proteins, including PP13, a member of the galectin family . Secreted by placental syncytiotrophoblasts, PP13 enters maternal circulation as early as the 5th week of gestation . In healthy pregnancies, PP13 plasma concentrations rise progressively throughout gestation, peaking at around 400 pg/mL by term . The PP13 content of the placenta is approximately 2.5 mg by the end of gestation, thereby constituting about 7% of placental proteins .
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p2
|
PMC11276665
|
sec[0]/p[2]
|
1. Introduction
| 4.367188 |
biomedical
|
Study
|
[
0.99951171875,
0.00038361549377441406,
0.00021147727966308594
] |
[
0.87744140625,
0.0017242431640625,
0.12005615234375,
0.0005445480346679688
] |
The significance of PP13 lies in its association with preeclampsia (PE) , a hypertensive disorder of pregnancy, characterized by onset in the first trimester and often leading to maternal and perinatal morbidity and mortality . Research indicates a stark decrease in PP13 levels in the maternal circulation during the first trimester of pregnancies complicated by P. This is in contrast to healthy pregnancies, where concentrations steadily increase . Altered levels of PP13 mRNA and protein, as well as polymorphic variants found in women with PE, suggest a potential role for PP13 in the pathogenesis of this disorder . PE poses significant risks to maternal and fetal health, including increased cardiovascular disease risk and improper vascular remodeling, which can lead to intrauterine growth restriction. Correct vascular remodeling during pregnancy is essential for maintaining adequate utero-placental blood flow, with the uterine circulation undergoing expansive remodeling to accommodate the increased demand for oxygen and nutrients required for fetal growth .
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p3
|
PMC11276665
|
sec[0]/p[3]
|
1. Introduction
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
0.0002884864807128906,
0.0002377033233642578
] |
[
0.7734375,
0.004413604736328125,
0.2218017578125,
0.0005345344543457031
] |
PP13 modulates several key processes that are critical for ensuring a successful pregnancy, including maternal immune responses to prevent maternal–fetal rejection , modulate oxidative stress and inflammation to mitigate oxidative damage and inflammatory responses , promote cytotrophoblast invasion into the myometrium, and ensure spiral artery remodeling . Animal studies have also shown that PP13 modulates endothelial cell function , promotes angiogenesis, and facilitates the vascular remodeling necessary for proper placental and fetal development .
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p4
|
PMC11276665
|
sec[0]/p[4]
|
1. Introduction
| 4.082031 |
biomedical
|
Study
|
[
0.99951171875,
0.0002391338348388672,
0.0001163482666015625
] |
[
0.998046875,
0.00033283233642578125,
0.0016689300537109375,
0.00010251998901367188
] |
Despite these findings, our understanding of how PP13 affects the maternal vasculature during pregnancy is incomplete. Therefore, the objective of this study was to investigate the effects of PP13 on the vasculature of pregnant and non-pregnant women and to elucidate its mechanisms of action and potential implications in pregnancy-related vascular physiology. A comprehensive understanding of how PP13 affects maternal vessels is essential for elucidating the pathophysiology of pregnancy-related vascular disorders. This research may provide valuable insights into the diagnostic and therapeutic applications of PP13 in pregnancy-related disorders.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276665_p5
|
PMC11276665
|
sec[1]/p[0]
|
2. Results
| 4.121094 |
biomedical
|
Study
|
[
0.99951171875,
0.0002663135528564453,
0.00014913082122802734
] |
[
0.9990234375,
0.00022542476654052734,
0.0005130767822265625,
0.00008088350296020508
] |
PP13 and its vehicle were assessed for their effects on uterine arteries that were isolated from pregnant women. The results demonstrated a concentration-dependent vasodilatiory effect, with a maximal dilation of 14 ± 5.4% at a concentration of 10 −8 M. The EC 50 value for PP13 was calculated to be 1.7 × 10 −12 M. In contrast, the administration of the vehicle did not induce any significant effect on arterial diameter . These findings indicate the potent vasodilatory action of PP13 and highlight the biological activity of PP13 in regulating uterine vascular tone.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p6
|
PMC11276665
|
sec[1]/p[1]
|
2. Results
| 3.628906 |
biomedical
|
Study
|
[
0.9990234375,
0.0003008842468261719,
0.0006704330444335938
] |
[
0.99853515625,
0.0010080337524414062,
0.0003032684326171875,
0.00008338689804077148
] |
PP13 and its vehicle were also evaluated on subcutaneous arteries obtained from pregnant women. Unlike uterine arteries, neither PP13 nor its vehicle significantly affected subcutaneous arterial function . These results suggest the specificity of PP13 action on uterine arteries in this experimental context.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276665_p7
|
PMC11276665
|
sec[1]/p[2]
|
2. Results
| 4.140625 |
biomedical
|
Study
|
[
0.99951171875,
0.0002636909484863281,
0.00014090538024902344
] |
[
0.9990234375,
0.00023496150970458984,
0.000518798828125,
0.00008183717727661133
] |
Given the evidence for a vasodilatory role for PP13 in uterine arteries, we sought to investigate its potential counteractive effects against the potent vasoconstrictor U46619, a thromboxane receptor agonist. Uterine arteries, isolated from pregnant women, were exposed to U46619 in the presence and absence of PP13. As anticipated, U46619 induced a concentration-dependent contraction of uterine arteries. However, intriguingly, the contraction was significantly attenuated in the presence of PP13 . The maximum U46619-contraction was reduced by 71% (from 40 ± 6.3% to 12 ± 4.8%) by PP13 addition. These findings highlight the vasomodulatory potential of PP13 in regulating uterine vascular tone.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p8
|
PMC11276665
|
sec[1]/p[3]
|
2. Results
| 4.117188 |
biomedical
|
Study
|
[
0.99951171875,
0.0002658367156982422,
0.00015175342559814453
] |
[
0.9990234375,
0.00024187564849853516,
0.00045013427734375,
0.00007289648056030273
] |
To determine whether PP13’s effect on vascular tone is dependent on pregnancy, we conducted experiments using uterine arteries isolated from non-pregnant women. Interestingly, PP13 reduced U46619-induced contraction in these arteries as well, but the change was not significant, as depicted in Figure 4 . PP13 was less effective in attenuating U46619 contraction in non-pregnant women than in pregnant women. In uterine arteries taken from non-pregnant women, PP13 reduced the maximum U46619-contraction by only 32% (from 72 ± 9.2% to 49 ± 14.6%). Thus, its vasodilatory effect is potentiated in the pregnant state.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p9
|
PMC11276665
|
sec[1]/p[4]
|
2. Results
| 4.117188 |
biomedical
|
Study
|
[
0.99951171875,
0.0002359151840209961,
0.0001952648162841797
] |
[
0.99951171875,
0.00019478797912597656,
0.00034999847412109375,
0.00006008148193359375
] |
To elucidate the mechanism by which PP13 counteracts U46619-induced contraction in uterine arteries isolated from pregnant women, we investigated whether PP13 acts through the nitric oxide pathway. To this end, uterine arteries were preincubated with specific inhibitors of the enzyme NOS, L-NAME, and L-NNA and then exposed to U46619 in either the presence or absence of PP13 . Although the concentration–response curve for U46619 contraction remained significantly ( p < 0.05) reduced, the counteracting effects of PP13 were much lower than those reported in Figure 3 ( p < 0.01).
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p10
|
PMC11276665
|
sec[1]/p[5]
|
2. Results
| 4.136719 |
biomedical
|
Study
|
[
0.99951171875,
0.0002300739288330078,
0.0001628398895263672
] |
[
0.99853515625,
0.0009708404541015625,
0.0005025863647460938,
0.00010663270950317383
] |
This indicates that the inhibition of NO production in part prevents the observed antagonistic action of PP13 on U46619-induced contraction and suggests that PP13 may exert its vasomodulatory effects by modulating the production or availability of nitric oxide, most likely via the endothelium.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p11
|
PMC11276665
|
sec[1]/p[6]
|
2. Results
| 4.140625 |
biomedical
|
Study
|
[
0.99951171875,
0.0002532005310058594,
0.00019252300262451172
] |
[
0.99951171875,
0.0002110004425048828,
0.000347137451171875,
0.00006395578384399414
] |
To further elucidate the transduction pathway through which PP13 exerts its effects, we investigated its action in the presence of a specific inhibitor of guanylate cyclase, ODQ. Uterine arteries were preincubated with ODQ to inhibit the production of the nucleotide cGMP. In the presence of ODQ, the dose–response curve for U46619 contraction in PP13 treated vessels remained significantly reduced ( p < 0.01), as shown in Figure 6 . These results suggest that PP13’s mechanism of action is cGMP-independent and most likely affects vascular smooth muscle.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p12
|
PMC11276665
|
sec[2]/p[0]
|
3. Discussion
| 4.171875 |
biomedical
|
Study
|
[
0.99951171875,
0.0003192424774169922,
0.00016021728515625
] |
[
0.9990234375,
0.00020170211791992188,
0.0005602836608886719,
0.00008541345596313477
] |
This study investigated the impact of PP13 on human vasculature, focusing specifically on both uterine (reproductive) and subcutaneous (systemic) arteries isolated from pregnant and non-pregnant women. The results revealed several key findings: (1) PP13 exhibited pregnancy-specific vasodilatory action in uterine arteries, while no such effect was observed in subcutaneous arteries, indicating the selectivity of PP13 action on the reproductive vasculature; (2) PP13 mitigated the vasoconstriction induced by U46619, which was particularly pronounced during pregnancy; and (3) this was achieved partially via its stimulation of the NO pathway.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p13
|
PMC11276665
|
sec[2]/p[1]
|
3. Discussion
| 4.042969 |
biomedical
|
Study
|
[
0.99951171875,
0.0001863241195678711,
0.00024056434631347656
] |
[
0.9990234375,
0.0007266998291015625,
0.0003559589385986328,
0.00006318092346191406
] |
This study was performed on small subcutaneous and uterine arteries with a lumen diameter of less than 250 μm. These vessels are recognized as pivotal regulators of vascular resistance, thereby controlling blood flow to downstream organs and tissues. Specifically, uterine arteries are responsible for supplying blood to the uterus, a role which becomes even more critical during pregnancy as they orchestrate circulation to the feto-placental unit.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
PMC11276665_p14
|
PMC11276665
|
sec[2]/p[2]
|
3. Discussion
| 4.230469 |
biomedical
|
Study
|
[
0.99951171875,
0.00037980079650878906,
0.0003249645233154297
] |
[
0.8056640625,
0.0023441314697265625,
0.19140625,
0.00046443939208984375
] |
It is well established that profound adaptations must occur in the structure and function of the maternal uterine circulation during pregnancy to sustain the metabolic demands of fetal development and optimize utero-placental blood flow . Regulated by the complex interplay of hormonal, mechanical, and paracrine factors, these include angiogenesis, growth, vasodilation, and alterations in vascular compliance and tone . Together, these changes lead to the substantial and progressive enhancement of utero-placental blood flow, which increases approximately 10–20 times above non-pregnant uterine flow levels. Studies utilizing Doppler ultrasound and MRI have reported uterine blood flow values of around 50 mL/min in non-pregnant women, which escalate to levels of 500–600 mL/min during late pregnancy . This significant increase underscores the dynamic nature of the tissue and the vascular adaptations necessary to sustain fetal growth and development.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p15
|
PMC11276665
|
sec[2]/p[3]
|
3. Discussion
| 4.167969 |
biomedical
|
Study
|
[
0.99951171875,
0.0001773834228515625,
0.00013494491577148438
] |
[
0.99755859375,
0.00080108642578125,
0.0016422271728515625,
0.00009399652481079102
] |
The placenta assumes a central role in orchestrating physiological changes within the maternal uterine vasculature during pregnancy through its secretion of hormones, growth factors, and cytokines that exert endocrine and paracrine effects . Given the marked decrease it induces in the plasma concentration in PE , evaluating the effects of PP13 on the maternal vasculature is paramount. Notably, preeclampsia is linked to impaired uterine vascular remodeling, resulting in elevated vascular resistance and reduced utero-placental blood flow .
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p16
|
PMC11276665
|
sec[2]/p[4]
|
3. Discussion
| 4.078125 |
biomedical
|
Study
|
[
0.99951171875,
0.00018596649169921875,
0.00014030933380126953
] |
[
0.99853515625,
0.0006241798400878906,
0.0007948875427246094,
0.00008368492126464844
] |
The effects of PP13 on subcutaneous and uterine arteries were assessed using pressure myography, a well-established ex vivo technique renowned for its ability to closely replicate in vivo physiological conditions. This method, which allows for the examination of vascular responses in the presence of a true transmural pressure, is more reflective of the in vivo situation than traditional wire myography.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p17
|
PMC11276665
|
sec[2]/p[5]
|
3. Discussion
| 4.117188 |
biomedical
|
Study
|
[
0.99951171875,
0.00018990039825439453,
0.00017571449279785156
] |
[
0.99951171875,
0.00023090839385986328,
0.0003333091735839844,
0.000056684017181396484
] |
Our findings underscored the specificity of PP13 action on uterine arteries. This aligns with PP13’s role as a placental protein that is primarily involved in reproductive processes. Although prior studies demonstrated PP13-induced vasodilation in rat uterine arteries, as well as in mesenteric arteries , this study is the first to examine PP13’s effects on human uterine arteries, a critical distinction given that preeclampsia occurs exclusively in humans.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999999 |
PMC11276665_p18
|
PMC11276665
|
sec[2]/p[6]
|
3. Discussion
| 4.382813 |
biomedical
|
Study
|
[
0.99951171875,
0.0002560615539550781,
0.0001531839370727539
] |
[
0.998046875,
0.0009741783142089844,
0.001003265380859375,
0.0001360177993774414
] |
Moreover, PP13’s ability to counteract U46619-induced vasoconstriction, which is particularly enhanced during pregnancy, may be facilitated by gestation-associated vascular conditions, such as increased endothelial NO levels in uterine arteries . The endothelial cells lining the lumen of all blood vessels, including uterine arteries, are surrounded by vascular smooth muscle cells (VSMCs). These endothelial cells serve as sensors and effectors of multiple inputs, such as pressure, flow (shear stress), and vasoactive substances present in the bloodstream. In turn, they release various mediators, including NO, prostaglandins, and other hyperpolarizing factors, which induce the relaxation of VSMCs (endothelium-dependent dilation).
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p19
|
PMC11276665
|
sec[2]/p[7]
|
3. Discussion
| 4.097656 |
biomedical
|
Study
|
[
0.99951171875,
0.00021529197692871094,
0.0001863241195678711
] |
[
0.9990234375,
0.0005640983581542969,
0.00048279762268066406,
0.0000908970832824707
] |
Consistent with previous findings in rats, our results indicate that the vasodilatory actions of PP13 in human vessels are mediated through NO, indicating likely endothelium-dependent action and that this effectively opposes the action of vascoconstrictor influences .
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p20
|
PMC11276665
|
sec[2]/p[8]
|
3. Discussion
| 4.175781 |
biomedical
|
Study
|
[
0.99951171875,
0.000274658203125,
0.0002434253692626953
] |
[
0.931640625,
0.0528564453125,
0.01486968994140625,
0.0006275177001953125
] |
During pregnancy, the expression of endothelial nitric oxide synthase (eNOS) undergoes significant upregulation, primarily from elevated estrogens and shear stress . This upregulation is pivotal for augmenting endothelial vasodilatory influence on the adjacent VSM. As a result, uterine vascular resistance is reduced, leading to enhanced blood flow to the uterus and the feto-placental unit.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p21
|
PMC11276665
|
sec[2]/p[9]
|
3. Discussion
| 4.320313 |
biomedical
|
Study
|
[
0.99951171875,
0.0002434253692626953,
0.00014138221740722656
] |
[
0.994140625,
0.000568389892578125,
0.005367279052734375,
0.00014853477478027344
] |
The effects of NO are conventionally attributed to the activation of soluble guanylyl cyclase, which leads to the formation of cyclic guanosine-3,5-monophosphate (cGMP). However, our findings indicate that PP13-NO-induced vasodilation operates through a pathway that is independent of cGMP. This suggests the existence of alternative mechanisms that contribute to the vasodilatory effects of nitric oxide beyond the traditional NO-cGMP pathway. Emerging evidence has shed light on these alternative signaling pathways. For instance, nitric oxide has been shown to directly stimulate potassium channels and calcium channels . These findings underscore the complexity of nitric oxide signaling and its diverse physiological implications, paving the way for further exploration into the intricate and interactive mechanisms underlying vascular regulation.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p22
|
PMC11276665
|
sec[2]/p[10]
|
3. Discussion
| 3.810547 |
biomedical
|
Study
|
[
0.99951171875,
0.00015842914581298828,
0.00028634071350097656
] |
[
0.998046875,
0.0005807876586914062,
0.0011730194091796875,
0.00007528066635131836
] |
Although this study provided some insight into the role of NO in PP13’s vasodilatory effect, it did not delve into the precise molecular mechanisms underlying this action and further investigations are warranted to elucidate the specific pathways involved. Another limitation to consider is that our studies focused on isolated arteries, which cannot fully replicate the complex vascular environment in vivo.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p23
|
PMC11276665
|
sec[2]/p[11]
|
3. Discussion
| 3.982422 |
biomedical
|
Study
|
[
0.99951171875,
0.0002658367156982422,
0.00011545419692993164
] |
[
0.990234375,
0.0028533935546875,
0.0067901611328125,
0.0002014636993408203
] |
In conclusion, the insights gained from this study offer valuable groundwork for future investigations into a potential therapeutic role for PP13 in treating preeclampsia, a condition that affects approximately 8% of women worldwide and is still without a definitive cure. Clinical studies are warranted to ascertain the efficacy and safety of PP13 administration in preeclamptic women, ensuring positive outcomes for both maternal and fetal health.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p24
|
PMC11276665
|
sec[3]/sec[0]/p[0]
|
4.1. Patient Recruitment and Selection
| 3.931641 |
biomedical
|
Study
|
[
0.9931640625,
0.0066070556640625,
0.0004074573516845703
] |
[
0.99755859375,
0.0017795562744140625,
0.0003046989440917969,
0.0004131793975830078
] |
This study involved recruiting patients undergoing cesarean delivery or hysterectomy, all within reproductive age (ranging from 20 to 45 years). A total of 17 patients participated, comprising 5 non-pregnant individuals and 12 pregnant patients with normal pregnancies. Patients were informed about the project upon hospitalization and provided with detailed explanations. Those who agreed to participate underwent a health assessment and provided informed consent. Ethical approval was obtained from the local Research Ethics Committee [approval number: Prot. 8177, 11 February 2022)] in adherence with the principles outlined in the Declaration of Helsinki. Exclusion criteria included twin pregnancies and pre-existing conditions, such as chronic arterial hypertension, autoimmune, renal, or hepatic diseases, ensuring that enrolled patients were free from debilitating pathologies.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p25
|
PMC11276665
|
sec[3]/sec[1]/p[0]
|
4.2. Samples Collection
| 3.988281 |
biomedical
|
Study
|
[
0.99951171875,
0.0005927085876464844,
0.0001386404037475586
] |
[
0.99658203125,
0.0027294158935546875,
0.0003993511199951172,
0.00023162364959716797
] |
Samples of subcutaneous fat and myometrium (~1 cm 3 ) were obtained from consenting healthy women at the Department of Gynecology and Obstetrics, Annunziata Hospital of Cosenza, Italy. Tissues were immediately immersed in cold HEPES–physiological salt solution (HEPES-PSS), kept on ice, and transported within 30 min to the vascular physiology laboratory for arteriole isolation.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p26
|
PMC11276665
|
sec[3]/sec[2]/p[0]
|
4.3. Vessel Preparation
| 4.03125 |
biomedical
|
Study
|
[
0.99853515625,
0.0012083053588867188,
0.0004930496215820312
] |
[
0.51318359375,
0.48291015625,
0.002361297607421875,
0.0018033981323242188
] |
In the laboratory setting, tissues were initially rinsed with fresh cold HEPES-PSS to ensure the removal of any blood residue from the sample surface. Subsequently, the tissues were delicately transferred into small glass dissecting dishes (90 mm) filled with cold HEPES-PSS solution. Once in the dish, they were carefully positioned at the bottom and secured in place using insect pins. The bottom of the dishes was covered with a smooth layer of silicone elastomer (Sylgard 184) to provide a stable working surface.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p27
|
PMC11276665
|
sec[3]/sec[2]/p[1]
|
4.3. Vessel Preparation
| 3.830078 |
biomedical
|
Study
|
[
0.9990234375,
0.00040912628173828125,
0.00037097930908203125
] |
[
0.93115234375,
0.06683349609375,
0.001079559326171875,
0.000736236572265625
] |
Arterioles were dissected under a stereomicroscope (Leica MZ 12.5 Ergo) utilizing fine forceps (Dumont #55) and sharp Vannas spring scissors (Fine Science Tools, Foster City, CA, USA), ensuring the preservation of their structural integrity and minimizing any potential damage.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999997 |
PMC11276665_p28
|
PMC11276665
|
sec[3]/sec[3]/p[0]
|
4.4. Pressure Myography
| 4.113281 |
biomedical
|
Study
|
[
0.99951171875,
0.0003769397735595703,
0.0001811981201171875
] |
[
0.99755859375,
0.0020961761474609375,
0.0003294944763183594,
0.00013816356658935547
] |
Artery segments, cut free of surrounding adipose and connective tissue, were isolated and transferred to a pressure myograph chamber (Instrumentation and Model Facility, University of Vermont, Burlington, VT, USA). Cannulation was conducted at both ends of the vessel between two opposing glass cannulas filled with HEPES-PSS, requiring the use of forceps with fine tips to ensure precision. Arterial segments of a sufficient length, approximately 3 mm, were mounted to ensure that there was an undamaged segment between the two cannulas.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p29
|
PMC11276665
|
sec[3]/sec[3]/p[1]
|
4.4. Pressure Myography
| 4.199219 |
biomedical
|
Study
|
[
0.99951171875,
0.0005779266357421875,
0.0001499652862548828
] |
[
0.998046875,
0.0013380050659179688,
0.0005898475646972656,
0.00018703937530517578
] |
To elaborate on the cannulation process, one end of the vessel was mounted and tied onto glass pipettes using thin nylon suture thread, and any residual blood within the lumen was gently flushed out with the solution. Subsequently, the other end of the vessel was tied onto the downstream cannula, which was closed off to establish a “blind sack” preparation. Meanwhile, the inflow cannula, connected to a pressure-servo system (Living Systems Instrumentation, Burlington, VT, USA), was adjusted to a physiological pressure level of 50 mmHg to mimic in vivo conditions. This pressure-servo system was coupled with a pressure transducer to monitor the pressure applied to the vessels throughout the experiment. Each vessel was evaluated for leaks and, if present, discarded as these can influence vasodilation and vasoconstriction. The preparation was continuously superfused with HEPES-PSS, warmed to 37 °C and possessing a pH of 7. Arterial lumen diameter was continuously monitored using video microscopy, coupled with specialized data-acquisition software (Ionoptix 6.3.4.69, Westwood, MA, USA), to track the relative positions of the inner and/or outer vascular walls.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p30
|
PMC11276665
|
sec[3]/sec[4]/p[0]
|
4.5. Experimental Protocol
| 4.152344 |
biomedical
|
Study
|
[
0.99951171875,
0.00031256675720214844,
0.00018548965454101562
] |
[
0.99951171875,
0.00024962425231933594,
0.000377655029296875,
0.00007510185241699219
] |
Arteries were tested for viability using vasoactive agents U46619 (constrictor) and acetylcholine (dilator). Non-responsive vessels were excluded. Functioning arteries were equilibrated for 30–45 min and subjected to PP13 testing using two protocols: (1) vessels pre-constricted with U46619 were exposed to increasing PP13 concentrations (10 −13 –10 −8 M); and (2) a concentration–response curve for U46619 was generated in the absence vs. presence of PP13 (10 −8 M, pre-incubated for 1 h). At the end of the experiment, vessels were superfused with HEPES-PSS without calcium plus the phosphodiesterase inhibitor, papaverine (10 −4 M), to induce maximal vasodilation and to measure the maximal vessel diameter. The underlying molecular mechanism of PP13 action was investigated using the following pharmacological inhibitors: (1) Nω-nitro-L-arginine methylester (L-NAME 10 −4 M) and Nω-nitro-l-arginin (LNNA 10 −4 M) for the eNOS enzyme, and (2) 1H-oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) for the guanylate cyclase enzyme. Uterine arteries were preincubated with inhibitors for 20 min and then constricted with U46619 before undergoing testing with PP13.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999994 |
PMC11276665_p31
|
PMC11276665
|
sec[3]/sec[5]/p[0]
|
4.6. Chemicals and Solutions
| 3.908203 |
biomedical
|
Study
|
[
0.99951171875,
0.0001951456069946289,
0.0004074573516845703
] |
[
0.9833984375,
0.015960693359375,
0.00042700767517089844,
0.0001850128173828125
] |
The HEPES-PSS solution consisted of the following components in mM: 141.8 NaCl; 4.7 KCl; 1.7 MgSO 4 ; 0.5 EDTA; 2.8 CaCl 2 ; 10.0 HEPES; 1.2 KH 2 PO 4 ; 5.0 glucose. The solutions were prepared in deionized water and titrated with sodium hydroxide to a physiologic pH of 7. Chemicals were purchased from Sigma-Aldrich (Milan, Italy), Fisher Scientific (Milan, Italy), and Cayman Chemical Co. (Hamburg, Germany), unless otherwise specified. All drugs tested were administered from stock solutions, prepared daily, while PP13, U46619 and ODQ stock solutions were stored in aliquots and were thawed as needed.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999996 |
PMC11276665_p32
|
PMC11276665
|
sec[3]/sec[5]/p[1]
|
4.6. Chemicals and Solutions
| 1.742188 |
biomedical
|
Other
|
[
0.99267578125,
0.0008897781372070312,
0.006198883056640625
] |
[
0.38427734375,
0.6103515625,
0.00319671630859375,
0.0021343231201171875
] |
Recombinant PP13 was obtained from Protein Laboratories Hylabs, Ltd. (Rehovot, Israel). in collaboration with HyLaboratories Ltd. (Rehovot, Israel). The protein was dissolved in sterile water (vehicle).
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999998 |
PMC11276665_p33
|
PMC11276665
|
sec[3]/sec[6]/p[0]
|
4.7. Statistical Analyses
| 4.039063 |
biomedical
|
Study
|
[
0.99951171875,
0.0002391338348388672,
0.0001379251480102539
] |
[
0.9990234375,
0.0004277229309082031,
0.0004906654357910156,
0.00007593631744384766
] |
The vasodilatory effect of PP13 was expressed as a percentage of maximal diameter, which was determined in the presence of the relaxing solution. EC 50 was calculated as the concentration that induced 50% of the maximum response. The area under the concentration–response curve (AUC) was compared across experimental conditions. Data were expressed as means ± standard error mean (SEM), with statistical significance set at p < 0.05 using an unpaired Student’s t -test.
|
[
"Mariacarmela Gatto",
"Milena Esposito",
"Michele Morelli",
"Silvia De Rose",
"Sveinbjorn Gizurarson",
"Hamutal Meiri",
"Maurizio Mandalà"
] |
https://doi.org/10.3390/ijms25147522
|
N/A
|
https://creativecommons.org/licenses/by/4.0/
|
en
| 0.999995 |
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