Estrous Phase Research Articles

Expression of IRS2 in the female reproductive system during the estrous cycle in mice

ABSTRACT Insulin receptor substrate 2 (IRS2) participates in reproduction; however, the location and expression of IRS2 in the reproductive system of female mice is not clear. We used real-time quantitative polymerase chain reaction (RT-PCR), western blot and immunohistochemical staining to investigate the expression of IRS2 in the ovary, oviduct and uterus of female mice during the estrous cycle. We found that IRS2 was expressed in all reproductive organs of mouse and that the expression level changed with the estrous phases. The expression of IRS2 in reproductive organs was greatest during estrus.

Effect of Music Exposure on Sexual Behavior of Female Rats

Background: The human limbic system is excited by music. The nervous system sends music signals converted into neural information by inner hair cells to the primary auditory area and limbic system. This activates genes that play a role in social and cognitive function. This study aims to prove the effect of music on physiological sexual behavior. Methods: The study was conducted on adult female Rattus norvegicus rats aged 2-3 months with a body weight of about 200 grams. The rats were exposed to the sound of music which divided into soft (Javanese), medium (Balinese gamelan), and loud (Dangdut tekno) music categories. The duration of exposure for each type of music is two hours in a row for seven days. During music exposure, rats' behavior in cages was recorded using CCTV. The test carried out is a normality test using the T-test (student's t-test). Results: A comparison test between treatment groups showed that all groups did not show differences in general behavior. For sexual behavior, the normality test showed that the Balinese group was the only abnormal group. The Piano group's sexual behavior was statistically the same as the control group. Conclusion: Exposure to Balinese gamelan showed an increase in sexual activity in female rats. Balinese gamelan music shows a relationship between music and limbic system activity, which is indicated by increased activity in the estrous phase. Keywords: emotion, limbic system, music, sexual activity

The investigation of apelin and apelin receptor expressions in mouse endometrium during peri-implantation period

BackgroundFertilization, pre-implantation embryo development, implantation, and decidualization are critical for a healthy pregnancy. Successful implantation requires a competent blastocyst and a receptive uterus. Apelin was purified from the bovine stomach in 1998. Apelin receptor (APJ) is a member of G protein-coupled receptors. Apelin/APJ system’s physiological role was shown in cardiovascular system, immune response, stress response, fluid regulation, nutrient uptake, angiogenesis, and adipoinsular axis; however, whether apelin/APJ system plays a role in implantation is unknown. In our study, we aimed to evaluate the localization and expressions of the apelin/APJ system in the peri-implantation period mouse uterus. MethodsUteri and implantation sites were collected from mice on the estrous phase and the 1st, 4th, 5th, 6th, and 8th days of pregnancy. Also, inter-implantation sites were collected from the 5th day of the pregnancy group. Localization and expressions of apelin and APJ were determined by immunohistochemistry and Western blot, respectively. ResultsApelin and APJ were expressed in the luminal and gland epithelium, the stroma of all experimental groups. Two isoforms of apelin-8 and 16 kDa were detected by Western blot. While apelin expression increased from the estrous to the 8th day of pregnancy, APJ expression increased from the estrous to the 4th day of pregnancy, reached the highest expression level, then decreased. ConclusionsOur findings suggest that the apelin/APJ system might be involved in implantation and decidualization. Our findings will guide further studies and may help elucidate the underlying causes of implantation failure and pregnancy loss.

Evaluations of Ovarian and Luteal Blood Flow Waveform Patterns in Buffalos Subjected to OvSynch Protocol in Cold and Hot Seasons

This current study aimed to determine ovarian and luteal blood flow waveform patterns in buffalos synchronized using OvSynch protocol in cold and hot seasons. Six cyclic buffalo cows aged 6±0.5 years old, having a weight of 400 ± 50 kg, were scanned daily along three successive estrous cycles transrectally by Doppler ultrasonography to evaluate the normal ovarian hemodynamic during the normal spontaneous ovulation and before the start of experiments. Buffaloes were synchronized with gonadotropin[GnRH] –prostaglandin[P] –gonadotropin (GPG) protocol in which animals received 10μg of GnRH on day ??, 0.250μg of PGF2α on day 7, and another dose of 10μg of GnRH was administered 48h after the PGF2α injection. Blood sampling and ovarian ultrasound examinations (color and spectral Doppler modes) were conducted on the day of the estrous and luteal phases. Results revealed that peak systolic velocity waveform (PSV) was significantly (P<0.05) increased in the cold season compared to the hot season. The Luteal blood flow after the end of OvSynch protocol on days (5,7,9, and 11) was significantly increased in the cold season than that in the hot one. The serum levels of estradiol (E2) and nitric oxide (NO) after the second GnRH injection in the OvSynch protocol were significantly (P<0.05) elevated in the cold season as compared to the hot one.
 Moreover, the progesterone (P4) levels had risen in OvSynch-treated buffaloes on days 5,7,9, and 11 of the cycle in the cold season compared to the hot one. Conclusion: In the cold season, ovarian hemodynamics was significantly improved compared to the hot one; this may influence the reproductive efficiency of buffaloes. Further studies were needed to prove it.

5-HT1B receptor agonist enhances breakpoint for cocaine on a progressive ratio (PR) schedule during maintenance of self-administration in female rats but reduces breakpoint for sucrose.

Background: Previous research showed that the 5-HT1B receptor agonist CP94253 enhanced cocaine reinforcement rate during maintenance of daily self-administration (SA), but inhibited reinforcement rate after 21 days of abstinence in male rats. Here we examined whether female rats show similar effects of CP94253 during maintenance as males across estrous cycle phases. Methods: Female rats trained on a fixed ratio 5 (FR5) cocaine reinforcement schedule were tested for the effects of CP94253 (5.6 mg/kg, s.c.) on cocaine reinforcement rate during each phase of the estrous cycle, with access to either low (0.075 and 0.1875) or high (0.375 and 0.75) cocaine doses available for 1 h sequentially in descending dose order. Other female and male rats trained on a progressive ratio (PR) schedule of cocaine or sucrose reinforcement were tested for CP94253 (0, 3.2, 5.6, and 10 mg/kg, s.c.) effects on reinforcement rate in 3-h sessions. CP94253 effects on responding during sucrose cue-reactivity were also examined post-abstinence. Results: Regardless of sex, CP94253 enhanced breakpoints on the PR schedule during maintenance of cocaine SA but attenuated breakpoints for sucrose reinforcement and decreased responding during sucrose cue-reactivity. FR results showed that CP94253 attenuated cocaine reinforcement rate during all estrous cycle phases except metestrus. Conclusions: Overall, we suggest that CP94253 increased incentive motivation for cocaine during maintenance of SA in female and male rats, yet decreased motivation for sucrose. We also suggest that 5-HT1BRs modulate motivation similarly across sexes except when females are in metestrus.

Hypothalamic insulin expression remains unaltered after short-term fasting in female rats.

Our previous study showed that 6-h fasting increased insulin expression in the hypothalamus of male rats. We, therefore, wanted to examine if this phenomenon occurs in female rats and whether it depended on the estrus cycle phase. Female rats in proestrus or diestrus were either exposed to 6-h fasting or had ad libitum access to food. The serum, cerebrospinal fluid, and hypothalamic insulin levels were determined using radioimmunoassay. The hypothalamic insulin mRNA expression was measured by RT-qPCR, while the hypothalamic insulin distribution was assessed immunohistochemically. Albeit the short-term fasting lowered circulating insulin, both hypothalamic insulin mRNA expression and hypothalamic insulin content remained unaltered. As for the hypothalamic insulin distribution, strong insulin immunopositivity was noted primarily in ependymal cells lining the upper part of the third ventricle and some neurons mainly located within the periventricular nucleus. The pattern of insulin distribution was similar between the controls and the females exposed to fasting regardless of the estrous cycle phase. The findings of this study indicate that the control of insulin expression in the hypothalamus differs from that in the pancreatic beta cells during short-term fasting. Furthermore, they also imply that the regulation of insulin expression in the female hypothalamus is different from males but independent of the estrus cycle phase.

The Effect of Alternating High-Sucrose and Sucrose Free-Diets, and Intermittent One-Day Fasting on the Estrous Cycle and Sex Hormones in Female Rats.

Relationships between diet, sex hormone concentrations, and the estrous cycle are important from the perspective of infertility and estrogen-dependent disease prevention and treatment. Four dietary interventions reflecting modern eating behaviors were explored. The study involved 50 female rats divided into five feeding groups. The impact of the amount of sucrose consumed (9% and 18% of the dietary energy content), alternating high-sucrose and sucrose-free diets, and a high-sucrose diet combined with intermittent one-day fasting on the estrous cycle and sex hormone concentrations in female rats was assessed. Even low amounts of dietary sucrose (9% of the dietary energy content) were found to lead to increased estradiol (E2) concentrations and decreased progesterone (Pg) concentrations. A high-sucrose diet, even when periodically applied, additionally led to a reduced concentration of luteinizing hormone (LH). The largest changes in the hormones tested were observed with one-day fasting coupled with the high-sucrose diet; in addition, the estrous phase was shortened and the estrous cycle was disrupted. The results of this study show that both the amount of dietary sucrose and also its uptake pattern affect the estrous cycle and sex hormone concentrations in female rats.

Streptomycin treatment of genital carriers of Leptospira in experimentally infected sheep on different estrous phases

Leptospirosis in ruminants presents as a chronic disease that causes several reproductive disorders leading to severe economic losses. The current recommended treatment can be efficient to eliminate the renal carrier state, however little is known about the effect of this drug in removing the genital carrier state and the hormonal influence on it. A total of 12 primiparous sheep experimentally infected with a strain of Leptospira santarosai serogroup Sejroe, FV52 strain, were used and distributed as group A (estrus; n = 5), group B (metaestrus; n = 4) and group C (control; n = 3). At D0, groups A and B were treated with streptomycin (25 mg/kg) single dose. Samples of cervicovaginal mucus (CVM) were collected on days 0, 3, and 35 post-treatment, while uterine fragment (UF) samples were collected on days 3 and 35, for PCR. Even after antibiotic treatment, all groups presented infected animals, at D3 and D35, with no significant difference between the treated and control groups. Based on these results, it was conducted a second protocol of treatment with streptomycin, IM (25 mg/kg) for three consecutive days, which was 100% effective to eliminate the genital carrier state; therefore, that protocol should be recommended.

An Experimental Study- to Evaluate the Effect of Shatapushpa Taila on Estrous Phase of Female Wistar Albino Rats

From early adolescence to menopause, the menstrual cycle affects a woman’s physiological and psychological health for about 35 to 40 years in a normal population. Properly functioning hypothalamo pituitary ovarian axis, receptive endometrium with healthy ovum and sperm are considered to be the essential factors for conception. Acharyas have quoted Rutu, Kshetra, Ambhu and Beeja as Garbha sambhava samagri which is essential for conception where in Beeja can be taken as ovum. Shatapusha is mentioned in Kashyapa samhitha as Artavajanaka, hence an effort to evaluate its effect on estrous phase of female Wistar albino rats is the aim of this study. The selected animal was grouped into four groups with 6 animals each. The animal experiment showed significant increase in estrus phase of the oestrous cycle in rats which is the active phase of mating where the female receives the male.

Juvenile Exposure to BPA Alters the Estrous Cycle and Differentially Increases Anxiety-like Behavior and Brain Gene Expression in Adult Male and Female Rats

Perinatal exposure to bisphenol A (BPA) in murine models has been reported to affect social behavior and increase anxiety. However, there is little information about the effects of BPA exposure during puberty, a period in which sex hormones influence the maturation and differentiation of the brain. In this work, we evaluated the effect of BPA administration during the juvenile stage (PND 21–50) on anxiety in male and female rats. Newly weaned Wistar rats were treated with BPA (0, 50, or 500 µg/kg/day) for 30 days. To compare the intra- and inter-sex behavioral profiles, rats were evaluated using four different anxiety models: the Open field test (OFT), the Elevated plus maze (EPM), the Light-dark box test (LDBT), and the Defensive burying test (DBT). Males exhibited a clear-cut anxious profile at both doses in all four tests, while no clear behavioral effect of BPA exposure was observed in female rats. The latter showed an altered estrous cycle that initiated earlier in life and had a shorter duration, with the estrous phase predominating. Moreover, the expression of ESR1, ESR2, GABRA1, GRIN1, GR, MR, and AR genes increased in the hippocampus and hypothalamus of male rats treated with 50 µg/kg, but not in females. Our results indicate that BPA consistently induces a higher anxiety profile in male than in female rats, as evidenced predominantly by an increase in passive-coping behaviors and changes in brain gene expression, highlighting the importance of sex in peripubertal behavioral toxicology studies.

Endometrial gene expression in response to lipopolysaccharide between estrous cycle phases and uterine horns in cattle

Uterine bacterial community abundances shift throughout the estrous cycle, potentially altering the immunological environment of the uterus and impacting subsequent fertility. The objective of the current study was to evaluate the immunological impact of lipopolysaccharide (LPS), as a model for potentially pathogenic bacteria, throughout the uterine endometrium between the luteal and follicular phase of the estrous cycle. Bovine uterine tracts were harvested in mid-luteal (n = 7) or follicular (n = 7) phase. Explants were collected from the contralateral and ipsilateral horn relative to the dominant follicle or corpus luteum, then subjected to one of three treatments: uncultured control, cultured control, or cultured with LPS (1 µg/mL). Explants underwent RNA extraction and targeted RNA sequencing for expression analyses of 40 immune response related genes. Sequencing reads were mapped to Bos taurus genome in CLC Genomics Workbench. Resulting total read counts were normalized by housekeeping gene GAPDH and analyzed for overall expression profile by Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) and Variable Importance in Projection (VIP) analyses in Metaboanalyst. Individual gene expression differences were determined by GLIMMIX procedure in SAS with fixed effects of treatment, estrous phase, uterine horn, and their interaction, with random effect of individual uterus. Expression of 29 genes were affected among treatment groups, with seven genes increased in LPS treatment compared to other groups (P < 0.05). Multiple genes were affected by estrous phase and uterine horn, independent of treatment (P < 0.05). The OPLS-DA analyses indicated overall gene expression differences due to clustering by estrous cycle and treatment (P < 0.001), with no effect of uterine horn (P > 0.10). Similar clustering was observed between luteal and follicular phase explants of controls, but distinct separate clustering between phases with LPS treatment (P = 0.001). According to VIP analyses, mucins were identified as contributing the most to differences observed between phase and treatment. In conclusion, estrous cycle phase resulted in differing overall endometrial gene expression profiles of immune response to LPS treatment. Therefore, altered immunological environment of the uterus in response to bacteria at different estrous cycle stages may lead to differences in reproductive success.

Temporal endurance of exercise-induced benefits on hippocampus-dependent memory and synaptic plasticity in female mice

Exercise facilitates hippocampal neurogenesis and neuroplasticity that in turn, promotes cognitive function. Our previous studies have demonstrated that in male mice, voluntary exercise enables hippocampus-dependent learning in conditions that are normally subthreshold for long-term memory formation in sedentary animals. Such cognitive enhancement can be maintained long after exercise has ceased and can be re-engaged by a subsequent subthreshold exercise session, suggesting exercise-induced benefits are temporally dynamic. In females, the extent to which the benefits of exercise can be maintained and the mechanisms underlying this maintenance have yet to be defined. Here, we examined the exercise parameters required to initiate and maintain the benefits of exercise in female C57BL/6J mice. Using a subthreshold version of the hippocampus-dependent task called object-location memory (OLM) task, we show that 14d of voluntary exercise enables learning under subthreshold acquisition conditions in female mice. Following the initial exercise, a 7d sedentary delay results in diminished performance, which can be re-facilitated when animals receive 2d of reactivating exercise following the sedentary delay. Assessment of estrous cycle reveals enhanced wheel running activity during the estrus phase relative to the diestrus phase, whereas estrous phase on training or test had no effect on OLM performance. Utilizing the same exercise parameters, we demonstrate that 14d of exercise enhances long-term potentiation (LTP) in the CA1 region of the hippocampus, an effect that persists throughout the sedentary delay and following the reactivating exercise session. Previous studies have proposed exercise-induced BDNF upregulation as the mechanism underlying exercise-mediated benefits on synaptic plasticity and cognition. However, our assessment of hippocampal Bdnf mRNA expression following memory retrieval reveals no difference between exercise conditions and control, suggesting that persistent Bdnf upregulation may not be required for maintenance of exercise-induced benefits. Together, our data indicate that 14d of voluntary exercise can initiate long-lasting benefits on neuroplasticity and cognitive function in female mice, establishing the first evidence on the temporal endurance of exercise-induced benefits in females.

Effect of estrous cycle phases on gene expression in bovine oviduct epithelial cells.

Background and Aim:The oviduct environment is of particular importance because it is the site of fertilization and early embryo development. The oviduct, as a component of the reproductive system, responds to ovarian hormone (estradiol [E2] and progesterone [P4]) stimuli depending on the estrous cycle phase. This study aimed to elucidate the effect of estrous cycle phases (follicular and early and late luteal phases) on gene expression patterns in bovine oviduct epithelial cells (BOECs).Materials and Methods:Oviducts were obtained from healthy slaughterhouse animals, corresponding to ipsilateral ovaries with dominant follicles or corpus luteum during early and late luteal phases. BOECs were recovered from the isthmus (IST) and ampulla (AMP), and the expression patterns of genes related to cytokinesis and mitosis mechanisms (rho-associated coiled-coil containing protein kinase and cellular communication network factor 2 [CCN2]), growth factors (insulin-like growth factor-binding protein 3, epidermal growth factor receptor [EGFR], vascular endothelial growth factor A, and EGFR), antioxidant mechanisms (glutathione peroxidase 4 [GPX4]), apoptosis (B-cell lymphoma 2), complement component (C3), energy metabolism (aldose reductase gene family 1-member b1 [AKRIB1] and solute carrier family 2), hormone receptors (estrogen receptor 1 and luteinizing hormone/choriogonadotropin receptor), and specific glycoproteins (oviductal glycoprotein 1) were analyzed.Results:High P4 levels (late luteal phase) affected the expression of important genes related to antioxidant mechanisms (GPX4), energy metabolism (AKRIB1), growth factors (IGBP3 and EGFR), and cell growth regulation (CCN2) in the AMP. Low P4 levels (early luteal phase) affected the expression of AKR1B1, IGBP3, and CCN2. In addition, estrogen likely had an effect on OVPGP expression in the cattle oviduct.Conclusion:Differential gene expression patterns of BOECs in the AMP during the luteal phase (antioxidant mechanisms, energy metabolism, growth factors, and immunological regulators) and in the IST during the follicular phase (glycoproteins) may influence their renewal and population proportions, modulating the oviduct environment as well as gamete and embryo physiology.

Molecular Mechanism of Equine Endometrosis: The NF-κB-Dependent Pathway Underlies the Ovarian Steroid Receptors' Dysfunction.

Endometrosis is a frequently occurring disease decreasing mares’ fertility. Thus, it is an important disease of the endometrium associated with epithelial and stromal cell alterations, endometrium gland degeneration and periglandular fibrosis. Multiple degenerative changes are found in uterine mucosa, the endometrium. However, their pathogenesis is not well known. It is thought that nuclear factor-κB (NF-κB), a cell metabolism regulator, and its activation pathways take part in it. The transcription of the profibrotic pathway genes of the NF-κB in fibrotic endometria differed between the follicular (FLP) and mid-luteal (MLP) phases of the estrous cycle, as well as with fibrosis progression. This study aimed to investigate the transcription of genes of estrogen (ESR1, ESR2) and progesterone receptors (PGR) in equine endometria to find relationships between the endocrine environment, NF-κB-pathway, and fibrosis. Endometrial samples (n = 100), collected in FLP or MLP, were classified histologically, and examined using quantitative PCR. The phase of the cycle was determined through the evaluation of ovarian structures and hormone levels (estradiol, progesterone) in serum. The transcription of ESR1, ESR2, and PGR decreased with the severity of endometrial fibrosis and degeneration of the endometrium. Moreover, differences in the transcription of ESR1, ESR2, and PGR were noted between FLP and MLP in the specific categories and histopathological type of equine endometrosis. In FLP and MLP, specific moderate and strong correlations between ESR1, ESR2, PGR and genes of the NF-κB pathway were evidenced. The transcription of endometrial steroid receptors can be subjected to dysregulation with the degree of equine endometrosis, especially in both destructive types of endometrosis, and mediated by the canonical NF-κB pathway depending on the estrous cycle phase.

The investigation of hippo signaling pathway in mouse uterus during peri-implantation period.

Events in the uterus during the peri-implantation period include embryo development, acquisition of uterine receptivity, implantation and decidualization. Hippo signaling pathway regulates cell proliferation, apoptosis and differentiation. We aimed to determine localization and expressions of pYAP (Phospho Yes-associated protein), YAP (Yes-associated protein), TEAD1 (TEA domain family member 1) and CTGF (Connective tissue growth factor), members of the Hippo signaling pathway, in the mouse uterus during the peri-implantation period. Pregnant mice were randomly separated into 5 groups: 1st, 4th, 5th, 6th, and 8th days of pregnancy groups. Non-pregnant female mice in estrous phase were included in the estrous group. Uteri and implantation sites were collected. Also, inter-implantation sites were collected from the 5th day of pregnancy group. pYAP, YAP, TEAD-1 and CTGF were detected by immunohistochemistry and Western blotting. We observed that the expressions of YAP, TEAD-1 and CTGF were increased in the luminal and glandular epithelium on the 1st and 4th days of pregnancy when epithelial proliferation occurred. pYAP expression was high, and YAP and CTGF expressions were low in the luminal epithelium of the implantation sites on the 5th day of pregnancy, when epithelial differentiation occurred. pYAP expression was low, YAP and CTGF expressions were high at implantation sites on the 6th and 8th days of pregnancy, where decidua was formed. Our findings suggest that the Hippo signaling pathway might be involved in implantation and decidualization. Our findings will guide further studies and may help to elucidate underlying causes of implantation failure and pregnancy loss.

The role of PACAP/PAC1R in PTSD: effects on fear extinction via the ventromedial hypothalamus

Introduction The incidence and severity of posttraumatic stress disorder (PTSD) is higher in women than men because of environmental and biological factors. Specific mechanisms in the PACAP-PAC1R (pituitary adenylate cyclase-activating polypeptide and its type I receptor) system may confer PTSD risk in women. Interestingly, while the PACAP (ADCYAP1) - PAC1R (ADCYAP1R1) system is expressed highly in the hypothalamus, no relationship has been described for this pathway in the hypothalamus with fear processing or in PTSD.ObjectivesWe studied whether the estrous/menstrual cycle at the moment of trauma predicts PTSD and the involvement of the PACAP neurons in the amygdala and hypothalamus during traumatic stress.MethodsMice: DREADDs, immunohistochemistry and behavior. Humans: fear-potentiated startle and questionnaires.Results Here, we show that acute stress immobilization (IMO) produces fear extinction impairments in female mice. Also, IMO elicits Adcyap1 and Adcyap1r1 mRNA upregulation in the hypothalamus, PACAP/c-Fos downregulation in the medial amygdala (MeA), and PACAP/FosB/ΔFosB upregulation in the ventromedial hypothalamus dorsomedial part (VMHdm) after fear extinction. We also found that women with the risk genotype of ADCYAP1R1 rs2267735 SNP show impaired fear extinction. In mice, DREADD-mediated inhibition of the MeA neurons projecting to the VMHdm during IMO rescues both PACAP upregulation in VMHdm and the fear extinction impairment. We ruled out contributions from inherent hormonal states showing that the menstrual or estrous cycle phase at the moment of trauma does not result in a vulnerable phenotype.ConclusionsOur data suggest that the PACAP-PAC1R hypothalamic system may be a novel candidate to treat and prevent PTSD symptoms including fear dysregulations.DisclosureNo significant relationships.

Citologia vaginal em éguas nas diferentes fases do ciclo estral associadas as estações do ano

The reproductive biotechnologies directly assist in the development of different methods of reproductive evaluation. The objective of this study was to develop a new method to determine the estrous cycle phase in equine females through vaginal cytology throughout the seasons, as well as to establish an estimate of inflammatory cells present in the vagina. Six mares were evaluated at different ages without a defined breed and reproductive activity and were subjected to ultrasound evaluation of the reproductive system and cytological analysis of the vaginal region. In the summer, there was a predominance of keratinized epithelial cells in the estrus and intermediate in the diestrus. In the autumn, there was a predominance of keratinized cells in estrus and parabasal cells in estrus and diestrus. In anestrus (winter), a greater number of parabasal cells was identified in relation to the other cell types. In estrus (spring), there was a predominance of parabasal and intermediate cells. Conversely, in diestrus, parabasals were found in greater numbers. The evaluation of inflammatory cells of the vaginal epithelium of mares showed greater activity in the summer; however, there are no reference values for healthy animals in the literature, and it is necessary to conduct studies on the subject. It is concluded that there is an influence of cyclicity on the vaginal epithelium of the equine species, varying according to the season. Additionally, vaginal cytological evaluation is an important complementary tool in the diagnosis of vaginitis in mares, requiring further research on the subject.

STREStrogen: The Dichotomous Role of Estrous Cycle on Anxiety‐like Behavior is Dependent upon Stress History

Stress‐induced psychiatric disorders are some of the most debilitating conditions worldwide and occur more often in women than men. Increased risk coincides with the reproductive years when circulating estrogen (E) levels peak, alluding to a critical role of ovarian hormones in susceptibility. Human stress studies support this association as E is linked to negative mood and anxiety during social stressors. However, preclinical research often reports opposing findings of protective effects of E in non‐stressed subjects, indicating reduced anxiety‐like behavioral responses. Yet, rodent studies often neglect to acknowledge the latent effects of stress on E‐induced behavioral differences, like the negative impact of a traumatic experience that perpetuates future interactions. Thus, this divergence between clinical and preclinical findings limits the capability of identifying adequate pharmacological targets for use in naturally cycling females. Therefore, we investigated if prior exposure to stress in intact adult female rats sensitized behavioral outcomes during anxiogenic tasks and whether this was contingent upon the current phase of estrous. Rats were exposed for 15 mins/day over 5 consecutive days to either control handling (CON) or witness stress (WS), a modified social defeat paradigm whereby a female observes social defeat from within a protected compartment of the aggressive resident’s cage. Forty‐eight hours after the 5th and final CON/WS exposure, rats underwent elevated plus maze (EPM) testing, and then acoustic startle response (ASR) testing the next day. Vaginal lavage was obtained immediately following EPM and ASR to confirm estrous phase. Analyses were conducted in a 2 x 2 fashion: stress exposure x estrous phase, in which metestrus/diestrus (M/D) signified putatively low levels of E, and proestrus/estrus (P/E) signified high levels. Results revealed a prominent stress exposure x estrous phase interaction for percentage of time spent in the closed and open arms of EPM. First, CON rats demonstrated reduced avoidance of the open arms when tested during P/E vs M/D. This is in line with studies reporting an anxiolytic effect of E. However, our findings in WS rats supports the notion that E is a critical component of sensitizing anxiety‐like responses in females with a history of stress. Specifically, WS rats demonstrated greater avoidance of open arms in the EPM during P/E vs M/D. Importantly, we also found that not all tests are impacted by cycle stage. Females with a history of stress demonstrated a sensitized startle amplitude compared with CON rats, independent of estrous phase, when tested for magnitude of the innate startle response. Further investigations into E receptor expression and subsequent signaling mechanisms in stress‐sensitive brain regions are necessary to determine how a history of stress modifies behavioral outcomes in the presence of high circulating E. These data can then elucidate novel therapeutic targets important for decreasing the high incidence of psychological disorders in this susceptible population.

Social Isolation Influences Endocrine and Metabolic Parameters in Female Mice Depending on Estrous State

Social distancing is a critical public health strategy employed to reduce the transmission of respiratory illnesses, such as COVID‐19. Studies conducted throughout the pandemic suggest that social distancing measures, including staying at home, simultaneously lead to an increased experience of isolation and loneliness. Social isolation is of potential concern because previous research in both humans and non‐human animals demonstrates that reduced social interaction has detrimental effects on mental and physical wellness, including an increase in mortality rate. Despite this background, less is known about how social isolation affects physiological parameters in a comprehensive manner, especially in females. We therefore conducted this study to investigate the extent to which social isolation simultaneously impacts multiple physiological and behavioral variables related to stress, metabolism, and reproduction. To do this, we housed three‐week‐old female Swiss‐Webster mice individually or in groups of three for five weeks. We measured body weight and food consumption each week of the experiment. During the last two weeks of treatment, we tracked the estrous cycle, using vaginal cytology to determine estrous states. After four weeks of housing treatment, we used closed system respirometry to measure oxygen consumption rate as an approximation of whole‐body metabolic rate. At the conclusion of the study, we collected blood in order to measure leukocyte counts, blood glucose concentration, and serum estrogen and progesterone concentrations. Individually housed females consistently consumed more food compared to group‐housed females. While there was no effect of social isolation on the length or proportion of time spent in the different estrous phases, the effects of isolation on other physiological variables depended on estrous state. Our preliminary results suggest that body weight, oxygen consumption rate, number of large lymphocytes, blood glucose concentration, and serum progesterone concentration were higher in individually housed females in diestrus, compared to group‐housed females in diestrus. In contrast, serum estrogen concentration was lower in individually housed females in diestrus, compared to group‐housed females in diestrus. Taken together, these findings suggest that social isolation has a substantial body‐wide effect on physiology and health in females, potentially mediated through a direct effect on ovarian hormone secretion. Furthermore, these findings highlight the importance of studying how stressors impact female physiology in naturally cycling animals.

Sex differences in the acute ethanol effects on object recognition memory: influence of estrous cycle.

Effects of acute ethanol (EtOH) on memory depend on several factors, including type of behavioral task. Sex differences in EtOH effects have been reported in humans and animals, and recognition memory can be influenced by circulating sex hormones. The aim of this study was to investigate the influence of sex and estrous cycle in the acute effects of EtOH on novel object recognition memory in rats. Male and female Wistar rats were part of one of the groups: control, 0.6-g/kg EtOH and 1.8-g/kg EtOH (administered intraperitoneally before the training session). The estrous cycle was evaluated by vaginal smear. The task was conducted in an open field arena. During training, animals were exposed to two identical objects, and test sessions were performed 1 h (short-term) and 24 h (long-term) later. One of the objects was changed in each test. Increased novel object exploration was shown by male and female controls in the short- and long-term tests, respectively. In the short-term test, females did not show preference for the novel object, and EtOH 1.8 g/kg impaired performance in males. In the long-term test, both sexes showed object discrimination, and 1.8-g/kg EtOH reduced preference for the new object in male rats. The phase of the cycle, the performance on proestrus was worse compared with other phases, and EtOH failed to impair performance mainly on estrous. In conclusion, while male rats displayed ethanol-induced recognition memory deficit, female rats were unaffected by EtOH impairing effects. In addition, the performance of female rats was influenced by the estrous cycle phases.