The impact of chronic restraint stress on the estrous cycle in NMRI female mice

Introduction: Stress represents a set of reactions in the organism activated by external factors. In order to maintain homeostasis and protect the organism, numerous mechanisms for adaptation to stress evolved. Stressors that act in short-term period cause acute stress reaction with generally positive effect on organism. When the stressor persists, and the organism fails to respond to the challenge, chronic stress develops, leading to pathological conditions, such as women's menstrual cycle disorders. Aim: To examine the impact of chronic restraint stress on the estrous cycle in NMRI female mice. Material and methods: A number of 12 mature female NMRI mice were randomly divided into control (n = 6) and experimental (n = 6) group. The induction of stress was performed for the experimental group by using restrain chambers 2 hours daily in 14 days. From the beginning of the experiment, vaginal lavages were taken from all mice for making smears that were analyzed to determine estrous cycle stages. For analysis of chronic stress effect, the frequency of estrus stages alternation observed in experimental group compared to control was examined. Results: The estrous cycle was observed and divided into proestrus, estrus, metestrus and diestrus. During the adaptation period, prolonged diestrus was dominantly present in both groups. When stress was induced, in the experimental group animals the absence of diestrus stage and oscillation to other stages was obseved, in contrast to the control group, where the diestus stage was frequently observed. Frequency of diestrus stage deviation in stressed animals was shown to be statistically significant (p < 0.01) when compared to control. Conclusion: The induction of chronic restraint stress in female NMRI mice leads to the alternation of the estrous cycle. Considering the fact that NMRI female mice share the similar hormonal regulation of the estrous cycle with women's menstrual cycle, they could present a solid model for studying women's reproductive disorders.

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Chronic restraint stress impairs cognition via modulating HDAC2 expression

Translational Neuroscience ◽

10.1515/tnsci-2020-0168 ◽

2021 ◽

Vol 12 (1) ◽

pp. 154-163

Author(s):

Jie Wu ◽

Cui Liu ◽

Ling Zhang ◽

Bing He ◽

Wei-Ping Shi ◽

...

Keyword(s):

Chronic Stress ◽

Hippocampal Neurons ◽

Restraint Stress ◽

Glucocorticoid Receptors ◽

Chronic Restraint Stress ◽

Akt Signaling Pathway ◽

Object Recognition Test ◽

Protein Levels ◽

Synaptic Functions ◽

Plus Maze

Abstract Background To investigate the effects of chronic restraint stress on cognition and the probable molecular mechanism in mice. Methods In the current work, a restraining tube was used as a way to induce chronic stress in mice. The protein levels were determined with ELISA and western blot. A series of behavior tests, including the Morris water maze, elevated plus maze, open field test, and novel object recognition test, were also performed to examine the anxiety and the ability of learning and memory. Moreover, murine neuroblastoma N2a cells were used to confirm the findings from mice under chronic stress. Results Decreased synaptic functions were impaired in chronic stress with the downregulation of PSD95, GluR-1, the neurotrophic factor BDNF, and immediate-onset genes Arc and Egr. Chronic restraint decreased the histone acetylation level in hippocampal neurons while HDAC2 was increased and was co-localized with glucocorticoid receptors. Moreover, chronic stress inhibited the PI3K/AKT signaling pathway and induced energy metabolism dysfunctions. Conclusion This work examining the elevated levels of HDAC2 in the hippocampus may provide new insights and targets for drug development for treating many neurodegenerative diseases.

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LIMK1/2 in the mPFC Plays a Role in Chronic Stress-Induced Depressive-Like Effects in Mice

The International Journal of Neuropsychopharmacology ◽

10.1093/ijnp/pyaa067 ◽

2020 ◽

Vol 23 (12) ◽

pp. 821-836

Author(s):

Ting-Ting Gao ◽

Yuan Wang ◽

Ling Liu ◽

Jin-Liang Wang ◽

Ying-Jie Wang ◽

...

Keyword(s):

Chronic Stress ◽

Restraint Stress ◽

Social Defeat ◽

Mild Stress ◽

Chronic Unpredictable Mild Stress ◽

Chronic Restraint Stress ◽

Lim Domain ◽

Social Defeat Stress ◽

Chronic Social Defeat Stress ◽

Pharmacological Target

Abstract Background Depression is one of the most common forms of mental illness and also a leading cause of disability worldwide. Developing novel antidepressant targets beyond the monoaminergic systems is now popular and necessary. LIM kinases, including LIM domain kinase 1 and 2 (LIMK1/2), play a key role in actin and microtubule dynamics through phosphorylating cofilin. Since depression is associated with atrophy of neurons and reduced connectivity, here we speculate that LIMK1/2 may play a role in the pathogenesis of depression. Methods In this study, the chronic unpredictable mild stress (CUMS), chronic restraint stress (CRS), and chronic social defeat stress (CSDS) models of depression, various behavioral tests, stereotactic injection, western blotting, and immunofluorescence methods were adopted. Results CUMS, CRS, and CSDS all significantly enhanced the phosphorylation levels of LIMK1 and LIMK2 in the medial prefrontal cortex (mPFC) but not the hippocampus of mice. Administration of fluoxetine, the most commonly used selective serotonin reuptake inhibitor in clinical practice, fully reversed the effects of CUMS, CRS, and CSDS on LIMK1 and LIMK2 in the mPFC. Moreover, pharmacological inhibition of LIMK1 and LIMK2 in the mPFC by LIMKi 3 infusions notably prevented the pro-depressant effects of CUMS, CRS, and CSDS in mice. Conclusions In summary, these results suggest that LIMK1/2 in the mPFC has a role in chronic stress-induced depressive-like effects in mice and could be a novel pharmacological target for developing antidepressants.

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Vitamin D Regulatory Effect on Restraint Stress-Induced Changes on Serum Corticosterone and Hippocampus BDNF Levels in Male Rats

Middle East Journal of Rehabilitation and Health ◽

10.5812/mejrh.115164 ◽

2021 ◽

Vol In Press (In Press) ◽

Author(s):

Katayoun Sedaghat ◽

Sara Choobdar ◽

Ahmad Reza Bandegi ◽

Zahra Ghods

Keyword(s):

Vitamin D ◽

Chronic Stress ◽

Restraint Stress ◽

Male Rats ◽

Negative Effects ◽

Bdnf Level ◽

Serum Corticosterone ◽

Protein Levels ◽

Induced Changes ◽

The Impact

Background: Chronic stress exerts negative effects on cognitive functions through inducing changes in the hippocampus. Brain-derived neurotrophic factor (BDNF) is an essential factor in cognitive activities, which is considerably reduced under chronic stress. 1,25(OH)2 vitamin D plays neuroprotective roles partially by regulating the expression of various neurotrophic factors. Objectives: Since few studies have studied the impact of vitamin D on BDNF level, we conducted this brief experiment to understand the role of vitamin D in maintaining hippocampal BDNF protein levels by using restraint as a model of chronic stress in rats. Methods: Rats underwent restraint stress 3 h/day for 28 days, during which they received vitamin D (5, 10 μg/kg) or its vehicle (IP, twice weekly). After the stress period, serum corticosterone (CORT) and hippocampus BDNF protein levels were measured. Results: Restraint stress increased serum CORT (P < 0.001) and reduced BDNF protein levels (P < 0.001) as compared to the non-stress group. Vitamin D markedly maintained BDNF level close to normal (P < 0.001), but did not change CORT level significantly. Conclusions: This study demonstrated that 3h/day of chronic restraint stress for 28 days boosted serum CORT and declined hippocampal BDNF levels, similar to stronger restraint stress models. Vitamin D maintained BDNF level close to normal in the hippocampus, but it did not affect CORT level significantly.

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Cardiac contraction, calcium transients, and myofilament calcium sensitivity fluctuate with the estrous cycle in young adult female mice

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00730.2013 ◽

2014 ◽

Vol 306 (7) ◽

pp. H938-H953 ◽

Cited By ~ 19

Author(s):

Jennifer K. MacDonald ◽

W. Glen Pyle ◽

Cristine J. Reitz ◽

Susan E. Howlett

Keyword(s):

Estrous Cycle ◽

Action Potentials ◽

Ventricular Myocytes ◽

Fold Increase ◽

Reproductive Hormones ◽

Cardiac Contraction ◽

Estrous Cycles ◽

Female Mice ◽

Ec Coupling ◽

The Impact

This study established conditions to induce regular estrous cycles in female C57BL/6J mice and investigated the impact of the estrous cycle on contractions, Ca2+ transients, and underlying cardiac excitation-contraction (EC)-coupling mechanisms. Daily vaginal smears from group-housed virgin female mice were stained to distinguish estrous stage (proestrus, estrus, metestrus, diestrus). Ventricular myocytes were isolated from anesthetized mice. Contractions and Ca2+ transients were measured simultaneously (4 Hz, 37°C). Interestingly, mice did not exhibit regular cycles unless they were exposed to male pheromones in bedding added to their cages. Field-stimulated myocytes from mice in estrus had larger contractions (∼2-fold increase), larger Ca2+ transients (∼1.11-fold increase), and longer action potentials (>2-fold increase) compared with other stages. Larger contractions and Ca2+ transients were not observed in estrus myocytes voltage-clamped with shorter action potentials. Voltage-clamp experiments also demonstrated that estrous stage had no effect on Ca2+ current, EC-coupling gain, diastolic Ca2+, sarcoplasmic reticulum (SR) Ca2+ content, or fractional release. Although contractions were largest in estrus, myofilament Ca2+ sensitivity was lowest (EC50 values ∼1.15-fold higher) in conjunction with increased phosphorylation of myosin binding protein C in estrus. Contractions were enhanced in ventricular myocytes from mice in estrus because action potential prolongation increased SR Ca2+ release. These findings demonstrate that cyclical changes in reproductive hormones associated with the estrous cycle can influence myocardial electrical and contractile function and modify Ca2+ homeostasis. However, such changes are unlikely to occur in female mice housed in groups under conventional conditions, since these mice do not exhibit regular estrous cycles.

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Chronic restraint stress triggers dopaminergic and noradrenergic neurodegeneration: Possible role of chronic stress in the onset of Parkinson’s disease

Brain Behavior and Immunity ◽

10.1016/j.bbi.2015.08.015 ◽

2016 ◽

Vol 51 ◽

pp. 39-46 ◽

Cited By ~ 40

Author(s):

Shuei Sugama ◽

Kazunari Sekiyama ◽

Tohru Kodama ◽

Yoshiki Takamatsu ◽

Takato Takenouchi ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Chronic Stress ◽

Restraint Stress ◽

Chronic Restraint Stress

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Chronic Mild Stress Causes Bone Loss via an Osteoblast-Specific Glucocorticoid-Dependent Mechanism

Endocrinology ◽

10.1210/en.2016-1658 ◽

2017 ◽

Vol 158 (6) ◽

pp. 1939-1950 ◽

Cited By ~ 5

Author(s):

Holger Henneicke ◽

Jingbao Li ◽

Sarah Kim ◽

Sylvia J. Gasparini ◽

Markus J. Seibel ◽

...

Keyword(s):

Bone Loss ◽

Chronic Stress ◽

Structural Parameters ◽

Chronic Mild Stress ◽

Bone Resorption Marker ◽

Mild Stress ◽

Male Mice ◽

Female Mice ◽

Glucocorticoid Signaling ◽

The Impact

Abstract Chronic stress and depression are associated with alterations in the hypothalamic–pituitary–adrenal signaling cascade and considered a risk factor for bone loss and fractures. However, the mechanisms underlying the association between stress and poor bone health are unclear. Using a transgenic (tg) mouse model in which glucocorticoid signaling is selectively disrupted in mature osteoblasts and osteocytes [11β-hydroxysteroid-dehydrogenase type 2 (HSD2)OB-tg mice], the present study examines the impact of chronic stress on skeletal metabolism and structure. Eight-week-old male and female HSD2OB-tg mice and their wild-type (WT) littermates were exposed to chronic mild stress (CMS) for the duration of 4 weeks. At the endpoint, L3 vertebrae and tibiae were analyzed by micro–computed tomography and histomorphometry, and bone turnover was measured biochemically. Compared with nonstressed controls, exposure to CMS caused an approximately threefold increase in serum corticosterone concentrations in WT and HSD2OB-tg mice of both genders. Compared with controls, CMS resulted in loss of vertebral trabecular bone mass in male WT mice but not in male HSD2OB-tg littermates. Furthermore, both tibial cortical area and area fraction were reduced in stressed WT but not in stressed HSD2OB-tg male mice. Osteoclast activity and bone resorption marker were increased in WT males following CMS, features absent in HSD2OB-tg males. Interestingly, CMS had little effect on vertebral and long-bone structural parameters in female mice. We conclude that in male mice, bone loss during CMS is mediated via enhanced glucocorticoid signaling in osteoblasts (and osteocytes) and subsequent activation of osteoclasts. Female mice appear resistant to the skeletal effects of CMS.

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Spilanthes acmella Murr. ameliorates chronic stress through improving mitochondrial function in chronic restraint stress rats

Neurochemistry International ◽

10.1016/j.neuint.2021.105083 ◽

2021 ◽

pp. 105083

Author(s):

Wilasinee Suwanjang ◽

Waralee Ruankham ◽

Banthit Chetsawang ◽

Sujira Mukda ◽

Sukhonthar Ngampramuan ◽

...

Keyword(s):

Chronic Stress ◽

Mitochondrial Function ◽

Restraint Stress ◽

Chronic Restraint Stress ◽

Spilanthes Acmella

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Jie-Yu-He-Huan Capsule Ameliorates Anxiety-Like Behaviours in Rats Exposed to Chronic Restraint Stress via the cAMP/PKA/CREB/BDNF Signalling Pathway

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/1703981 ◽

2021 ◽

Vol 2021 ◽

pp. 1-19

Author(s):

Xiwen Geng ◽

Hongyun Wu ◽

Zifa Li ◽

Chuanfen Li ◽

Dan Chen ◽

...

Keyword(s):

Chronic Stress ◽

Hpa Axis ◽

Restraint Stress ◽

Critical Factor ◽

Receptor Expression ◽

Monoamine Oxidase A ◽

Chronic Restraint Stress ◽

Elevated Plus Maze Test ◽

Anxiolytic Drugs ◽

Bdnf Pathway

Chronic stress is a critical factor in the aetiology of anxiety disorders; however, in the clinic, enduring and preventive measures are not available, and therapeutic drugs are associated with inevitable side effects. Our study established an anxiety rat model using chronic restraint stress (CRS) and assessed these animals using the open-field test, elevated plus-maze test, and light-dark box test. Jie-Yu-He-Huan capsule (JYHH), a Chinese medicine formula, was used as a preventative drug. The HPA axis-mediated release of corticotropin-releasing hormone, adrenocorticotropic hormone, and corticosterone from the hypothalamus was tested. In the hippocampus and prefrontal cortex, concentrations of 5-HT and its metabolite 5-hydroxyindoleacetic acid, as well as monoamine oxidase A, glucocorticoid receptor, and 5-HT1A receptor expression levels, were measured. Furthermore, we examined protein and mRNA expression of cAMP-PKA-CREB-BDNF pathway components. The results showed that JYHH had a significant preventative effect on the anxiety-like behaviour induced by CRS and prevented abnormal changes in the HPA axis and 5-HT system. Furthermore, CRS inhibited the cAMP-PKA-CREB-BDNF pathway, which returned to normal levels following JYHH treatment. This might be the underlying molecular mechanism of the antianxiety effect of JYHH, which could provide a new clinical target for preventative anxiolytic drugs for chronic stress.

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Plasma and ovarian metabolomics responses to chronic stress in female mice

10.1101/2022.01.03.474852 ◽

2022 ◽

Author(s):

Oana A Zeleznik ◽

Tinayi Hunag ◽

Chirag J Patel ◽

Elizabeth M Poole ◽

Clary B Clish ◽

...

Keyword(s):

Amino Acids ◽

Chronic Stress ◽

Restraint Stress ◽

Multiple Testing ◽

Ovarian Tissue ◽

Enrichment Analysis ◽

Rank Test ◽

Tissue Samples ◽

Female Mice ◽

Metabolite Set Enrichment Analysis

Background: Chronic stress may affect metabolism of amino acids, lipids, and other small molecule metabolites, but these alterations may differ depending on tissue evaluated. We examined metabolomic changes in plasma and ovarian tissue samples from female mice due to chronic stress exposure. Methods: At 12 weeks old, healthy, female, C57 black mice were randomly assigned to three weeks of chronic stress using daily restraint (2 hours/day; n=9) or normal care (n=10). Metabolomic profiling was conducted on plasma and ovarian tissues. Using the Wilcoxon Rank Test, Metabolite Set Enrichment Analysis, and Differential Network Analysis we identified metabolomic alterations occurring in response to restraint stress. All p-values were corrected for multiple testing using the false discovery rate approach. Results: In plasma, individual lysophosphatidylcholines (positively) and the metabolite classes carnitines (positively), diacylglycerols and triacylglycerols (inversely) were associated with restraint stress (adjusted-p's<0.2). In contrast, diacylglycerols and triacylglycerols were increased while carnitines were decreased in ovarian tissue from stressed mice (adjusted-p's<0.2). However, several metabolites (cholesteryl esters, phosphatidylcholines/ phosphatidylethanolamines plasmalogens and multiple amino acids) were consistently inversely associated with restraint stress in plasma and ovarian tissue (adjusted-p's<0.2). Conclusion: We identified differences in multiple lipid and amino acid metabolites in plasma and ovarian tissue of female mice after exposure to chronic stress. Some affected metabolites (primarily triacylglycerols and diacylglycerols) exhibited opposite associations with chronic stress in plasma (a marker of systemic influences) versus in ovarian tissue (representing local changes), suggesting research to understand the biological impact of chronic stress needs to consider both systemic and tissue-specific alterations.

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Role and Mechanism of Chronic Restraint Stress in Regulating Energy Metabolism and Reproductive Function Through Hypothalamic Kisspeptin Neurons

10.21203/rs.3.rs-622884/v1 ◽

2021 ◽

Author(s):

Yin-qiong Huang ◽

Junping Wen ◽

Gang Chen

Keyword(s):

Weight Loss ◽

Energy Metabolism ◽

Glucocorticoid Receptor ◽

Restraint Stress ◽

Reproductive Function ◽

Control Group ◽

Chronic Restraint Stress ◽

Stress Group ◽

Female Mice ◽

Significant Difference

Abstract Background: Stress activates the hypothalamic-pituitary-adrenal (HPA) axis, affecting energy homeostasis and reproductive function. The hypothalamic Kisspeptin neurons might be a new important central target in stress affecting energy metabolism and reproductive function.The aim of this study is to investigate whether stress affected energy metabolism and reproductive function through the glucocorticoid receptor on Kisspeptin neurons in the hypothalamus. Methods: There were four groups, that were control group, chronic restraint stress group (stress group), Kisspeptin specific glucocorticoid receptor knock out group (KGRKO group) and KGRKO+stress group. Body weight, food intake, the estrous cycle of female mice, serum sex hormone levels, serum corticosterone and prolactin, Kisspeptin expression in the hypothalamus were measured. Results: The restraint stress group showed a significant weight loss compared with the control group. Compared with the restraint stress group, the KGRKO+restraint stress group had a reduced weight loss, suggesting that restraint stress might partially affect the energy metabolism through GR on Kisspeptin neurons. In terms of reproductive function, the restraint stress group and the KGRKO+restraint stress group showed missing pre-estrus period or prolonged estrous cycles. Serum LH and FSH in KGRKO + restraint stress group decreased significantly compared with KGRKO group. However, no significant difference in the level of serum testosterone was observed. After restraint stress, the levels of serum cortisol and prolactin in male and female mice were significantly higher than the control group, and the hypothalamus Kiss1 gene mRNA expression and Kisspeptin protein expression were significantly decreased. Conclusion: Chronic restraint stress induced weight loss in mice. Chronic restraint stress played a negative role in regulating reproductive function. The effects of chronic restraint stress on energy metabolism and reproduction were partially mediated by glucocorticoid receptor on Kisspeptin neurons in the hypothalamus.

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