Estrous Phase Research Articles

Effect of Cucurbita pepo (Pumpkin) seed extracts on the cyclicity and reproductive hormones of female Wistar rats

Introduction: Despite the traditional use of Cucurbita pepo seed in pregnancy, its effects on female reproduction remain scarce. This study evaluated the impacts of n-hexane, dichloromethane (DCM), and aqueous ethanol extracts of C. pepo seed on the cyclicity and reproductive hormones of female Wistar rats. Methods: Ten groups of four rats received seed extracts or tween 80 orally for 21 days: A (control)= 0.5 mL tween 80 (vehicle); B, C, & D= 142.86, 285.71, and 428.57 mg/kg nHE; E, F, & G= 142.86, 285.71, 428.57 mg/kg of DCM; and H, I, & J= 142.86, 285.71, 428.57 mg/kg of aqueous ethanol extracts, respectively. Vaginal cytology monitored the estrous cycle daily, and blood samples were obtained for follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen, and progesterone at various oestrus cycle phases. Results: Compared to the control, the estrous cycle phases did not change significantly (P>0.05). FSH levels significantly increased (P<0.05) with DCM and aqueous ethanol extracts of C. pepo seed during proestrus and estrus phases compared to the control. A significant (P<0.05) increase in LH was observed with n-hexane, DCM, and aqueous ethanol extracts during all estrous cycle phases compared to the control. All extracts significantly increased estrogen levels (P<0.05) during all phases. DCM and aqueous ethanol extracts reduced substantially the estrus-phase progesterone. Conclusion: Cucurbita pepo may stimulate the hypothalamic-pituitary-gonadal axis in female reproduction. Further studies should be conducted using various phytoestrogen compounds to gain useful knowledge about the effectiveness, safety, and long-term effects of C. pepo seed extracts in regulating hormonal balance.

Mapping uterine calcium dynamics during the ovulatory cycle in live mice.

Uterine contraction patterns vary during the ovulatory cycle and throughout pregnancy, but prior measurements have produced limited and conflicting information on these patterns. We combined a virally delivered genetically encoded calcium reporter (GCaMP8m) and ultra-widefield imaging in live nonpregnant mice to characterize uterine calcium dynamics at organ scale throughout the estrous cycle. Prior to ovulation (proestrus and estrus), uterine excitations primarily initiated in a region near the oviduct, but after ovulation (metestrus and diestrus), excitations initiated at loci homogeneously distributed throughout the organ. The frequency of excitation events was lowest in proestrus and estrus, higher in metestrus, and highest in diestrus. These results establish a platform for mapping uterine activity and demonstrate that an anatomically localized trigger for uterine excitations depends on the estrous cycle phase.

Overestimation of sex differences in psychostimulant activity via comparisons of males and females from different behavioral groups.

There are inconsistencies in the observation of sex differences in baseline activity and psychostimulant activity. To address this, we have developed the MISSING (Mapping Intrinsic Sex Similarities as an Integral quality of Normalized Groups) model. MISSING model proposes that sex similarities are observed when we compare similar behavioral groups of males and females, with sex differences occurring when we compare distinct groups of sexes, but this model has not been tested. To test this model, we identified within-sex groups of Sprague Dawley rats (male n = 22, female n = 23) by conducted normal mixtures clustering of baseline activity, cocaine activity (as distance traveled in cm over 90 min) and cocaine activity normalized-to-baseline activity (NBA) of all subjects. We employed 2-way ANOVA to determine the impact of within-sex heterogeneity on sex differences. We compared our cluster-based method to current median-split approaches. Our new cluster-based method revealed three distinct clusters, each consisting of both males and females. We determined there were no sex differences in any of the variables when males and females from the same clusters were compared. The within-sex clusters for females were not defined by estrous phase. Median split analysis was ineffective in accurately identifying within-sex groups. Our results validate the MISSING model: there are no sex differences in psychostimulant activity except when we compare males and females from different behavioral groups. This has significant implications for how we proceed with research towards understanding the mechanism governing sex differences in psychostimulant activity.

Effect of the estrous cycle on zinc transporter expression in bovine cumulus-oocyte complexes and oviduct epithelial cells.

During the luteal and follicular phases of the estrous cycle, cumulus-oocyte complexes (COC) and oviduct epithelial cells (OEC) undergo notable physiological and morphological changes. Maintaining proper zinc (Zn) homeostasis is crucial in both somatic and germinal mammalian cells. This study aimed to assess the impact of the estrous phase (luteal or follicular) on Zn transporter expression in bovine COC and OEC (BOEC). The expression of Zn transporters Slc39a6 (ZIP6), Slc39a8 (ZIP8), Slc39a14 (ZIP14), Slc30a3 (ZnT3), Slc30a7 (ZnT7), and Slc30a9 (ZnT9) was analyzed in COC and BOEC from cows during the luteal or follicular phases. Gene expression of ZIP6, ZIP14, and ZnT9 was quantified in COC and BOEC. The gene expression in the remaining transporters could not be quantified due to low mRNA levels (ZIP8 and ZnT3 in COC and BOEC; ZnT7 in BOEC) or absence of expression (ZnT7 in COC). In COC, the relative expression (RE) of all three transporters was higher in the luteal phase compared to the follicular phase (P ≤ 0.05). In BOEC, the luteal phase increased the RE of ZIP 6 (P ≤ 0.05), decreased the RE of ZnT9 (P ≤ 0.05), and did not modify the RE of ZIP14 (P > 0.05) compared to the follicular phase. In conclusion, the study reveals differences in the gene expression of ZIP6, ZIP14, and ZnT9 according to the estrous cycle phase in ex vivo samples of bovine COC and OEC.

An Observation of the Microstructure of Cervical Mucus in Cows during the Proestrus, Estrus, and Metestrus Stages and the Impact on Sperm Penetration Ability.

Cervical mucus not only provides energy for sperm but also forms a barrier to block sperm. This paper aims to study the microstructure of cervical mucus in dairy cows during the proestrus, estrus, and metestrus and its effect on sperm permeability. The experiment collected cervical mucus from 60 Holstein cows during these phases, then observed the different shapes of the mucus after crystallization, classified the mucus, and analyzed its proportions. Scanning electron microscopy was used to observe the ultrastructure of the cervical mucus and measure the micro-pore sizes, followed by sperm permeability tests using mucus from different estrous stages and counting the number of permeated sperm. The results indicate that cervical mucus from cows in different estrous phases includes four types (L, S, P, G), with each type constituting a different proportion. During the proestrus, the L type was significantly more prevalent than the other types (p < 0.05); during estrus, the S type was significantly more prevalent than the other types (p < 0.05); and during the metestrus, the p type was significantly more prevalent than the other types (p < 0.05). The micro-pore sizes of the same type of cervical mucus did not show significant differences across different estrous phases (p > 0.05). However, within the same estrous phase, there were significant differences in the micro-pore sizes among the four types (p < 0.05). The number of sperm that permeated the cervical mucus during estrus and metestrus was significantly higher than during the proestrus (p < 0.05). This study provides data support for the research on cervical mucus in dairy cows.

Study of Ovary Follicular Profile and Estrogen Concentration of Peranakan Ongole (PO) Cows at Estrous Phase in Mountains and Lowland at Kulon Progo District, Yogyakarta

Study of Ovary Follicular Profile and Estrogen Concentration of Peranakan Ongole (PO) Cows at Estrous Phase in Mountains and Lowland at Kulon Progo District, Yogyakarta

Phytochemical analysis and effect of short-term administration of aqueous seed extract of Aframomum melegueta on haematologic indices of female albino rats

Background and aims: Aframomum melegueta (Alligator pepper) is a dietary spice widely used for entertainment, religious rites, food flavor enhancer, possessing erythropoietic potentials, and many other medicinal uses. Women are included in eating this widely used spice. This experiment determined the health risk or benefit of short-term administration of aqueous seed extract of A. melegueta on haematologic indices of female albino rats. Methods: Thirty adult female albino rats weighing 160 to 200 g were used for the study. Male rats were introduced into the female rat cages of groups II-V within 12h at the expected estrous phase for mating and withdrawn afterward. Groups I, II, and III were orally administered distilled water only, while groups IV and V received oral doses of 5 µg/kg b.w. Cabergoline and 3000 mg/kg b.w. aqueous seed extract of A. melegueta respectively 24 hours postpartum and once daily by oral gavage for three days. Results: Fourier-transform infrared spectroscopy (FTIR) identified twelve functional groups in the seed extracts, namely –OH, -NH2 , CH, -NH3 +, -CH3 , -OH, -N=C=O, -C≡N, -C=C=C, -NH, -CH3 and -1,3,5-trisubstituted benzenes, while gas chromatography with flame ionization detector (GCFID) determined 15 bioactive components namely kaempferol, naringenin, Sapogernin, flavanones, anthocyanin, flavan-3-ol, cyanogenic glycoside, ribalinidine, rutin, catechin, resveratrol, spartein, epicatechin, steroid and phytate. Non-significant alterations in hemoglobin, packed cell volume (PCV), and red blood cell count were observed. There was a significant (P&lt;0.05) decrease in WBC during pregnancy, but it was improved postpartum. However, platelet count was significantly (P&lt;0.05) reduced after extract administration. Conclusion: The results indicate no adverse anemic condition elicited during pregnancy and delivery on haematologic parameters, namely red blood cells, hemoglobin, and PCV, with a significant decrease in WBC during pregnancy as well as reduction in platelet counts (thrombocytopenia) of extract treated animals post-partum likely due to the flavonoid, resveratrol as well as rutin and (–)-epicatechin components of A. melegueta and thus may increase the risk of bleeding disorders but reflects a positive anti-atherogenic and cardioprotective effect.

Human CYP2D6 varies across the estrous cycle in brains of transgenic mice altering drug response

Cytochrome P450 (CYP) 2Ds are drug metabolizing enzymes found in brain and liver which metabolize numerous centrally acting drugs. Inhibition and induction of CYP2D-mediated metabolism in rodent brain alters brain drug and metabolite concentrations and resulting drug response. In female rats, brain CYP2D metabolism varies across the estrous cycle and with exogenous estrogen, changing brain drug concentrations and response.In this study harmine-induced hypothermia was lower in humanized CYP2D6 transgenic female mice during estrus compared to diestrus. Pretreatment into the cerebral ventricles with propranolol, a selective irreversible inhibitor of human CYP2D6 in brain, increased hypothermia in estrus but not in diestrus. In vivo enzyme activity was higher in brains of transgenic mice in estrus compared to diestrus and was lower after pretreatment with inhibitor in estrus, but not in diestrus. Hepatic activity and plasma harmine concentrations were unaffected by either estrous phase or inhibition of brain CYP2D6. In wild-type female mice, harmine-induced hypothermia was unaffected by either estrous phase or inhibitor pretreatment. Male mice were used as positive controls, where pretreatment with inhibitor increased harmine-induced hypothermia in transgenic but not wild-type, mice.This study provides evidence for female hormone cycle-based regulation of drug metabolism by human CYP2D6 in brain and resulting drug response. This suggests that brain CYP2D6 metabolism may vary, for example, during the menstrual cycle, pregnancy, or menopause, or while taking oral contraceptives or hormone therapy. This variation could contribute to individual differences in response to centrally acting CYP2D6-substrate drugs by altering local brain drug and/or metabolite concentrations.

The Influence of Sex Steroid Hormone Fluctuations on Capsaicin-Induced Pain and TRPV1 Expression.

Sexual dimorphism among mammals includes variations in the pain threshold. These differences are influenced by hormonal fluctuations in females during the estrous and menstrual cycles of rodents and humans, respectively. These physiological conditions display various phases, including proestrus and diestrus in rodents and follicular and luteal phases in humans, distinctly characterized by varying estrogen levels. In this study, we evaluated the capsaicin responses in male and female mice at different estrous cycle phases, using two murine acute pain models. Our findings indicate that the capsaicin-induced pain threshold was lower in the proestrus phase than in the other three phases in both pain assays. We also found that male mice exhibited a higher pain threshold than females in the proestrus phase, although it was similar to females in the other cycle phases. We also assessed the mRNA and protein levels of TRPV1 in the dorsal root and trigeminal ganglia of mice. Our results showed higher TRPV1 protein levels during proestrus compared to diestrus and male mice. Unexpectedly, we observed that the diestrus phase was associated with higher TRPV1 mRNA levels than those in both proestrus and male mice. These results underscore the hormonal influence on TRPV1 expression regulation and highlight the role of sex steroids in capsaicin-induced pain.

Moderate-intensity aerobic exercise enhanced dopamine signaling in diet-induced obese female mice without preventing body weight gain

Obesity continues to rise in prevalence and financial burden despite strong evidence linking it to an increased risk of developing several chronic diseases. Dopamine response and receptor density are shown to decrease under conditions of obesity. However, it is unclear if this could be a potential mechanism for treatment without drugs that have a potential for abuse. Therefore, the aim of this study was to investigate whether moderate-intensity exercise could reduce body weight gain and the associated decreases in dopamine signaling observed with high-fat diet-induced adiposity. We hypothesized that exercise would attenuate body weight gain and diet-induced inflammation in high-fat (HF)-fed mice, resulting in dopamine signaling (release and reuptake rate) comparable to sedentary, low-fat (LF)-fed counterparts. This hypothesis was tested using a mouse model of diet-induced obesity (DIO) and fast-scan cyclic voltammetry to measure evoked dopamine release and reuptake rates. Although the exercise protocol employed in this study was not sufficient to prevent significant body weight gain, there was an enhancement of dopamine signaling observed in female mice fed a HF diet that underwent treadmill running. Additionally, aerobic treadmill exercise enhanced the sensitivity to amphetamine (AMPH) in this same group of exercised, HF-fed females. The estrous cycle might influence the ability of exercise to enhance dopamine signaling in females, an effect not observed in male groups. Further research into females by estrous cycle phase, in addition to determining the optimal intensity and duration of aerobic exercise, are logical next steps.

The adipokine profile in the plasma and anterior pituitary of pigs during the estrous cycle

Adipokines play crucial roles in both reproductive and energy metabolic processes. This study aimed to compare the hormonal plasma profile of adiponectin, apelin, vaspin, chemerin, resistin, visfatin, and adipolin, and the expression of their receptors in the anterior pituitary (AP) between normal-weight Large White (LW) and fat Meishan (MS) pigs during different phases of the estrous cycle. We measured adipokine levels in the plasma and assessed their gene expression in the AP. We used Pearson’s correlation analysis to examine potential links between adipokines levels, their receptors, and metabolic parameters (body weight; backfat thickness) and reproductive parameters (pituitary weight; age at puberty; levels of gonadotropins, steroid hormones; and gene expression of gonadotropin-releasing hormone receptor and gonadotropins in AP). The plasma levels of the evaluated adipokines fluctuated with phase and breed, except for visfatin and adipolin. Moreover, adipokine expression in AP varied significantly between breeds and estrous cycle phases, except for resistin receptor CAP1. Notably, we observed a positive correlation between plasma levels of adiponectin and its transcript in the AP only in MS pigs. Apelin gene expression correlated negatively with its receptor in MS, while we observed a breed-dependent correlation between chemerin gene expression and its receptor CMKLR1. We identified significant positive or negative correlations between adipokines or their receptor levels in plasma and AP as well as metabolic or reproductive parameters, depending on the breed. In conclusion, we have demonstrated breed-specific and estrous cycle-dependent regulation of adipokines in AP, underscoring their potential impact on metabolic and reproductive processes in swine.

P-673 Tubulointerstitial nephritis antigen-like 1 (TINAGL1) expression is lower in the follicular fluid of patients with diminished ovarian reserve (DOR): A novel biomarker

Abstract Study question Are TINAGL1 levels in follicular fluid associated with ovarian reserve in patients undergoing ovarian stimulation for in-vitro fertilization (IVF)? Summary answer TINAGL1 concentration was lower in the follicular fluid (FF) of patients with DOR vs patients without infertility or polycystic ovarian syndrome (PCOS). What is known already TINAGL1 is an extracellular matrix protein involved in cell adhesion, proliferation, migration, and differentiation. TINAGL1 has previously been shown to be expressed in the mouse ovary and to be a serum biomarker that varies throughout the mouse estrous cycle, peaking in the estrous phase. Furthermore, TINAGL1 knockout mice have been shown to be infertile. TINAGL1 expression in the human ovary has not been previously evaluated. Study design, size, duration Between 2019 and 2023, a prospective study of 136 patients undergoing ovarian stimulation and oocyte retrieval (OR) was conducted at a single academic fertility center to assess TINAGL1 levels in luteinized FF and granulosa cells. Study groups were determined by infertility diagnosis: DOR n = 24, polycystic ovary syndrome (PCOS) n = 27, and control n = 85 (undergoing fertility preservation or had a partner with male factor only). Participants/materials, setting, methods TINAGL1 protein levels were quantified in the FF of patients that underwent OR via commercial enzyme-linked immunosorbent assay (ELISA). Granulosa cells were recovered from FF after OR, underwent lysis, RNA extraction, cDNA amplification and TINAGL1 mRNA expression was quantified via quantitative real-time polymerase chain reaction (qPCR). Demographics and infertility diagnoses were collected from the medical record. TINAGL1 levels were compared between the three groups. Statistical analysis was performed with parametric and non-parametric tests as appropriate. Main results and the role of chance Patients with DOR were younger when compared to the control and the PCOS group [Mean(SD) 32.7 (3.8) vs 34.45(3.9) vs 37.7 (3.0) years, p &amp;lt; 0.001] and had lower AMH levels [1.19 (0.8) vs 3.96 (3.03) vs 8.67 (4.56) ng/ml, p &amp;lt; 0.001]. TINAGL1 protein concentration in FF was measured by ELISA. Mean (SD) TINAGL1 levels were highest in PCOS, and lowest in the DOR group. Mean TINAGL1 levels were higher in the FF of the PCOS group when compared to the control group [4.73(7.15) vs 4.45 (4.57) ng/ml, p = 0.83] but this difference did not reach statistical significance. However, mean TINAGL1 FF levels were significantly lower in the DOR group when compared to the control group [2.54 (1.6) vs 4.48 (4.57) ng/ml, p = 0.04]. TINAGL1 was also found to be expressed in the granulosa cells with qPCR. TINAGL1 expression was upregulated by (+89%) in the PCOS group vs the control group (p = 0.0095). Importantly, there was a trend for decreased (-52%) TINAGL1 expression in DOR when compared to the control group (p = 0.67). The latter result should be interpreted with caution since only a subset of samples had sufficient granulosa cells for RNA extraction (DOR=4, control=18, PCOS=5). Limitations, reasons for caution This is a single institution study with a small number of patients in the DOR group (n = 24). Wider implications of the findings This is the first study to show that TINAGL1 is expressed in human granulosa cells. Decreased TINAGL1 may be a biomarker associated with DOR. Future studies should assess serum TINAGL1 levels at baseline and during ovarian stimulation and its contribution to the pathophysiology of DOR. Trial registration number Not applicable

P-649 Unconventional action of Luteinizing Hormone

Abstract Study question Has Luteinizing Hormone (LH) a role in the activation/survival of follicles of the primary ovarian reserve? Summary answer LH promotes survival and perhaps activation/growth of primordial and primary follicles, proliferation of granulosa cells of secondary follicles, reduces follicle atresia at later growing stages. What is known already In the mammalian ovary, Primordial Follicles (PMFs) constitutes the primary ovarian reserve, which declines over time due to physiological events such as atresia, recruitment, and ovulation. While in the adult, the role of LH in steroidogenesis and ovulation is well established, recent work from our laboratory has shown that under chemotherapeutic treatments, LH supports PMFs survival in prepuberal mouse ovary. Study design, size, duration LH effects on follicle dynamics and some activities of pre-granulosa cells were investigated in vitro and in vivo. In vitroexperiments were performed on PD4 GFP/Kit mice ovarian fragments and on PD8 CD1 mice ovarian somatic cells cultured up to six days in continuous presence of 0.2, 0.5, 1IU LH. In vivo experiments were performed on 9 PD4 and 8 PD28 CD1 mice receiving an intra-peritoneal injection for eight consecutive days of 1-5IU LH. Participants/materials, setting, methods Number and diameters of the GFP stained oocyte were counted in the ovarian fragments. Kit-Ligand and LH-R mRNA expression levels were analyzed by qPCR in pre-granulosa cells. For in vivo experiments, ovarian volume was measured by Micro-Ct and follicle counts were performed on histological sections. Estrous cycle phases were monitored by vaginal smear during the 8 days of treatment. Statistics was performed using Student’s t-test. Main results and the role of chance In fragments of PD4 mouse ovaries cultured in vitro for six days, 1IU LH significantly increased the number of oocytes with a diameter &amp;gt;40 mm (CTRL 17.7% vs LH 36.07%, p &amp;lt; 0,05), suggesting activation of PMFs. Moreover, the hormone increased the expression of mRNA for LHR and KL in the somatic ovarian cells and promoted their proliferation in vitro. The effects of 1IU LH on PD4 females can be summarized as follows: 1. Ovary volume increase (CTRL 0.3 mm3 vs LH 0.36 mm3, p = 0.07), 2. higher percent of PMFs (CTRL 68.227% vs LH 78.29%, p &amp;lt; 0,05) associated with lower percent of primary (CTRL 7.2% vs LH 4.05%, p &amp;lt; 0.04) and growing follicles (CTRL 14.18% vs LH 8.56%, p = 0.1), 3. higher number of granulosa cell layers (&amp;gt; three layers) in secondary follicles (CTRL 42.8% vs LH 54.6%), and 4. lower percent of atretic follicles (CTRL 2.64% vs LH 1.72%, p = 0.1). Similar results were obtained on 28 dpp mice. Morever, we observed that corpora lutea were present only in ovaries of LH-treated females and that all such females were in estrus. Limitations, reasons for caution The study was conducted only in mice. Wider implications of the findings These studies have highlighted new possible roles of LH on follicle classes (primordial/primary) whose development is considered gonadotropin independent. The results, if confirmed in humans, imply that LH might be used as a treatment for patients with ovarian disorders involved the survival/dynamics of the ovarian reserve. Trial registration number NA

Hormonal dependent expression of apelin and apelin receptor in the ovary and uterus of mice

Apelin and APJ have been shown to regulate female reproductive functions. However, its uterine expression during the oestrous cycle and its regulation by ovarian steroids, along with gonadotropin regulation in the ovary, has not been investigated. This study aimed to analyze the steroid-dependent uterine expression of apelin/APJ in the uterus along with the oestrous cycle. Furthermore, it also aimed to investigate gonadotropin-dependent ovarian expression of apelin and APJ. To investigate the uterine expression of apelin and APJ during estrous cycle in mice, uterus at different estrous stage were collected. To explore the ovarian steroids dependent expression of apelin system in the uterus, ovariectomized mice were treated with only estrogen at dose of 30 ng/g, only progesterone at dose of 150 μg/g and combined doses. To study the effect of gonadotropin on ovarian expression of apelin system, immature mice were injected with 2.5 IU of pregnant mare serum gonadotropin (PMSG) alone and both PMSG plus 2.5 IU of chorionic gonadotropin (hCG). Apelin and APJ protein expression are modulated by estrous phases in the uterus. The uterine apelin and APJ expression are up-regulated by estrogen and down-regulated by progesterone. The expression and localization of APJ showed increased abundance in the follicles of PMSG treated mice, however, the PMSG plus HCG treatment showed formation of corpus luteum with increased abundance of APJ and progesterone secretion. The expression of apelin and APJ are regulated by pituitary gonadotropin in the ovary and uterine apelin system by ovarian steroid hormone.

Dimorphic Regulation of the MafB Gene by Sex Steroids in Hamsters, Mesocricetus auratus.

MafB is a transcription factor that regulates macrophage differentiation. Macrophages are a traditional feature of the hamster Harderian gland (HG); however, studies pertaining to MafB expression in the HG are scant. Here, the full-length cDNA of the MafB gene in hamsters was cloned and sequenced. Molecular characterization revealed that MafB encodes a protein containing 323 amino acids with a DNA-binding domain, a transactivation domain, and a leucine zipper domain. qPCR assays indicated that MafB was expressed in different tissues of both sexes. The highest relative expression levels in endocrine tissues were identified in the pancreas. Gonadectomy in male hamsters was associated with significantly higher mRNA levels in the HG; replacement with dihydrotestosterone restored mRNA expression. The HG in male hamsters contained twofold more MafB mRNA than the HG of female hamsters. Adrenals revealed similar mRNA relative expression levels during the estrous cycle. The estrous phase was associated with higher mRNA levels in the ovary. A significantly up-regulated expression and sexual dimorphism of MafB was found in the pancreas. Therefore, MafB in the HG may play an active role in the macrophage differentiation required for phagocytosis activity and intraocular repair. Additionally, sex steroids appear to strongly influence the MafB expression in the HG and pancreas. These studies highlight the probable biological importance of MafB in immunological defense and pancreatic β cell regulation.

Gayo horse’s estrogen concentration after pospartum silent heat therapy using intramuscularly vitamin A, D, E

Abstract Silent heat is a condition in which livestock does not show symptoms of heat but physiologically estrus occurs due to low concentrations of the estrogen. Vitamins are small organic compounds that are needed by horses for the process of development, reproduction, and activity. Vitamin preparations are easily obtained and economically for breeders. This research aimed to observe differences in estrogen concentrations in postpartum silent heat gayo horses before and after therapy using intramuscularly vitamins A, D, E. Four of six gayo mares suspected of postpartum silent heat after diagnosis using ultrasound were used in this study. Four gayo mares were divided into two groups, the control group (K1, n=1) was only injected with NaCl and the therapy group (K2, n=3) was injected with vitamins A, D, E intramuscularly twice with an interval of one week. Blood plasma samples were collected before and after therapy for 7 days, then stored in the freezer at -20°C for analysis of estrogen concentrations using EIA. Data before and after therapy were analyzed using paired sample t-test and data between groups were reported descriptively. The results showed that the estrogen concentration of postpartum silent heat gayo horses in K1 was not significantly different (P&gt;0.05) before and after NaCl injection (251.71±57.90 vs 255.56±84.14 pg/ml), while there was a significant difference in K2 (P&lt;0.05) before and after vitamin A, D, E therapy (194.82±56.64 vs 557.11±96.55 pg/ml). Descriptively, there were differences in the concentration of the hormone estrogen in K1 and K2 after postpartum silent heat therapy. The conclusion is postpartum silent heat therapy using vitamins A, D, E in gayo horses can increase the concentration of the hormone estrogen in the estrous phase which has an impact on the appearance of estrus symptoms that is more clearly and easily observed by breeders.

The regulatory effect of electroacupuncture on endometrial M1-type macrophages in rats with intrauterine adhesions.

To observe the effect of electroacupuncture(EA) on endometrial fibrosis and M1-type macrophages in rats with intrauterine adhesions(IUA), so as to explore the possible mechanism of EA in the treatment of IUA. Fifteen female SD rats were randomly divided into blank group, model group and EA group, with 5 rats in each group. The IUA rat model was established by double damage method using mechanical scraping combined with lipopolysaccharide infection. Rats in the EA group were treated with acupuncture at "Guanyuan"(CV4), and EA at bilateral "Zusanli"(ST36) and "Sanyinjiao"(SP6)for 20 minutes each time, once a day, for 3 consecutive cycles of estrus. Five rats in each group were sampled during the estrous period, and the endometrial morphology, endometrial thickness and the number of blood vessels and glands were observed after HE staining. The fibrotic area of the uterus was observed after Masson staining. The positive expressions of Runt-related transcription factor(RUNX1), transforming growth factor-β1(TGF-β1), connective tissue growth factor(CTGF), α-smooth muscle actin(α-SMA), collagen type I(Col-Ⅰ), cluster of differentiation 86(CD86), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) in endometrial tissue were detected by immunohistochemistry. Western blot was used to detect relative protein expressions of RUNX1, TGF-β1, α-SMA, CD86, and TNF receptor 2 (TNFR2), and real-time fluorescence quantitative PCR was used to detect mRNA expressions of RUNX1, TGF-β1, α-SMA, CD86, and TNF-α in the endometrium. During the estrous phase, the endometrial layer in the model group was damaged, with reduced folds, disordered arrangement of epithelial cells, loose fibrous connective tissue, significant narrowing and adhesions in the uterine cavity, interstitial congestion, edema, and a significant infiltration of inflammatory cells with sparse glands. While uterine tissue structure of the EA group was basically intact, resembling a normal uterus, with more newly formed glands and a small amount of inflammatory cell infiltration. In comparison with the blank group, the endometrial thickness, the number of blood vessels, and the number of glands were significantly decreased(P<0.001) in the model group, while the ratio of uterine fibrosis area, the positive expressions of RUNX1, TGF-β1, CTGF, α-SMA, Col-Ⅰ, CD86, IL-1β, and TNF-α, the protein relative expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNFR2, and the mRNA relative expression levels of RUNX1, TGF-β1, α-SMA, CD86 and TNF-α in the endometrium were significantly increased (P<0.001, P<0.01). Compared to the model group, the endometrial thickness, the number of blood vessels, and the number of glands were significantly increased(P<0.01, P<0.05) in the EA group, while the ratio of uterine fibrosis area, the positive expressions of RUNX1, TGF-β1, CTGF, α-SMA, Col-Ⅰ, CD86, IL-1β and TNF-α in the endometrial tissue, the protein expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNFR2, and the mRNA relative expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNF-α in the endometrium were significantly decreased (P<0.001, P<0.01, P<0.05). EA can improve endometrial fibrosis in IUA rats, which may be related to its function in decreasing the level of endometrial M1-type macrophages and the secretion of related inflammatory factors.

Sex- Specific Effects of Chronic Unpredictable Stress on Renal Mitochondrial Function in C57BL/6 Mice

Background: The kidneys play a central role in long term control of blood pressure. Chronic psychological stress has been linked to hypertension, inflammation, and is associated with both an increased prevalence and progression of renal disease. However, the molecular and cellular mechanisms by which chronic stress alters renal function are not fully understood, and data on sex specific differences in renal and blood pressure responses to stress are limited. The impact of stress to cause local inflammation and mitochondrial dysfunction in the kidneys is not clear and represents a potential mechanism leading to renal dysfunction and hypertension. The chronic unpredictable stress (CUS) model induces psychological stress in rodents thus allowing the study of integrated physiological changes to the kidney in response to stress. Hypothesis: Exposing mice to CUS will lead to increased blood pressure that is associated with decreased renal mitochondrial function and renal inflammation. Methods: To investigate the link between stress, renal mitochondrial function, inflammation, and blood pressure, we exposed adult male and female C57BL/6 mice to CUS for 28 consecutive days. Blood pressure was measured via tail cuff on the 28th day of stress. 24 hours later, mice were euthanized and estrous cycle phase in females was determined by vaginal cytology. Kidney mitochondrial oxygen consumption was assessed using the Oroboros respirometer on the pole ends of the left kidneys. NLR family pyrin domain containing 3 (NLRP3) expression was measured in the right kidney using Real-Time TaqMan PCR. Results: Mean arterial pressure was elevated in both stressed males (in mmHg, 106±4 vs 87±4, n&lt;11, p&lt;0.01) and females (131± 6 vs 94±11 n&lt;4, p&lt;0.01) compared to non-stressed mice. While maximal electron transport capacity (ETS), a measure of mitochondrial function, was unchanged in CUS male kidneys, stressed females in the receptive (estrus and proestrus) phase of the estrous cycle exhibited a significant decrease in renal ETS compared to non-stressed females (in pmol O2/mg protein, 14.4±0.9 vs 11.4±0.7, n=11, p&lt;0.05). Renal mRNA expression of NLRP3 was increased in CUS males (35%± 7.7, n=8, p&lt;0.01) but unchanged in CUS females. Conclusion: These data suggest potential mechanistic differences in stress-induced hypertension in renal mitochondria (decreased ETS in stressed females) and inflammasome level (increased NLRP3 in stressed males) between male and female mice. This work was supported by P20GM109091 (F.H.), R01HL130972-01A1, R01HL5949, BX000168-10A1, BX005320 (F.G.S.), R01DK132948 (C.W. &amp; F.P.) R01 MH129798 (SKW); VA VISN7 RDA (F.H.),, 1U54HL169191-01 &amp; BX002604 (M.J.R.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the abstract. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The role of gender differences in the outcome of juvenile social isolation: Emphasis on changes in behavioral, biochemical and expression of nitric oxide synthase genes alteration

Social isolation can cause serious problem in performance of individuals in community. As gender differences may cause variation results in the severity of depressive behavior and response of patients to therapy, the impact of gender and the interaction of the level of endocrine secretion in depression were investigated in this study. Wistar rats of both sexes were subjected to post-weaning social isolation (PWSI) conditions and, together with the control group, experienced several behavioral tests including open-field Test (OFT), elevated plus maze (EPM), force swimming test (FST), splash test and novel object recognition test (NOR). Hippocampal tissue was isolated to measure biochemical factors such as nitric oxide level, FRAP amount, MDA level. In addition, real-time-PCR test was used to quantify the genes expression level of inducible nitric oxide synthase (iNOS) and neuronal nitric oxide synthase (nNOS). On the other hand, sexual hormone levels in blood were measured. Both cognitive and behavioral f unctions were declined as the result of PWSI induction in male and diestrus female rats. The consequent surge of estradiol during estrous phase seems to suppress the accumulation of reactive oxygen species (ROS), and modulate iNOS and nNOS expression. In conclusion, while the pattern of PWSI in surge cellular antioxidants, raising cellular ROS level is gender-specific, this alleviation was in relation with the drop of estradiol and unrelated with testosterone level.

Lactation suppression effect of aqueous seed extract of Aframomum melengueta on female albino rats

Background and aims: Alligator pepper, a dietary spice used as food flavor enhancer is a common ingredient in pepper soup, a spicy delight in most parts of West Africa and possesses both medicinal and nutritive properties. Despite the well documented benefits of breastfeeding for mothers and infants, there are a variety of medical situations for which breastfeeding is contraindicated. Therefore, lactation suppression might be indicated for the best interest of the mother and/or infant. This study was undertaken to investigate the Lactation suppression effect of aqueous seed extract of Aframomum Melengueta on female albino rats. Methodology: Thirty (30) adult female albino rats weighing 160 to 200 g were used for the study while eighteen (18) albino mice of body weights (b.w.) range of 25 g to 30 g were used for the preliminary acute toxicity test. Male rats were introduced into the female rat cages of groups II-V within 12h at the expected estrous phase for mating and withdrawn thereafter. Groups I, II and III were orally administered distilled water only, while Groups IV and V received oral doses of 5 µg/kg b.w. Carbagoline and 3,000 mg/kg b.w. aqueous seed extract of Aframomum melegueta respectively 24 hours postpartum once daily by oral gavage for 3 days. Results: LD50 determination revealed no death was recorded among the animals up to 5000 mg extract/kg body weight. Study indicates that Aframomum melengueta extract elicited significant reduction of prolactin with corresponding increase in estrogen and progesterone in postpartum animals. Histological sections of the mammary gland show extensive fibrosis and congested ducts lobular units within the tissue stroma of the mammary gland of Aframomum melengueta treated animals. Conclusion: The results confirms a positive lactation suppression effect of aqueous seed extract of Aframomum Melengueta on postpartum lactating female albino rats