Ovarian Activity Research Articles

Phytochemicals-based investigation of Rubia cordifolia pharmacological potential against letrozole-induced polycystic ovarian syndrome in female adult rats: In vitro, in vivo and mechanistic approach

Polycystic Ovarian Syndrome (PCOS) is a metabolic, reproductive, and endocrine disorder affecting women of fertile age. This study aimed to formulate a phytochemicals-based standardized aqueous ethanolic extract of Rubia cordifolia (SERC) to explore its pharmacological potential in PCOS-induced female rats and elucidate its mechanism. HPLC analysis revealed the presence of phytochemicals such as chlorogenic acid, p-coumaric acid, gallic acid, and kaempferol. Thirty female adult rats were divided into two groups for induction of PCOS (5 female rats in the normal control group + 25 female rats in the disease-induced group). PCOS was induced by administering letrozole (1 mg/kg p.o.) for 6 weeks. After PCOS induction, animals of the disease-induced group were divided into five groups: one group used as disease control (PCOS) group, one group on metformin (20 mg/kg), and three groups on SERC (200, 400, and 600 mg/kg). Histopathological analysis showed that PCOS induction reduced corpus luteum and developing follicles and increased cystic follicles. In comparison, SERC treatment improved ovulation with more primary and developing follicles. SERC reduced the serum insulin, LH surge, and testosterone levels while improving the FSH, estrogen, and progesterone serum levels. SERC significantly improved the oxidation status of the liver and normalized the lipid profile and liver function markers. In conclusion, SERC treated PCOS, and the suggested mechanism might be the restoration of aromatase activity and background inflammatory status improvement in ovaries.

Association between body condition profiles, milk production and reproduction performance in Holstein and Normande cows

Body-condition dynamics are known to affect the different steps of reproduction in cattle (cyclicity, estrus expression, fertilization, embryo development). This has led to a widespread idea that there is an ideal-target optimal body condition, but no clear profile has yet been identified. Here we investigated the relationships between body condition score (BCS) profiles and reproductive performance in dairy cows. Data were from Holstein or Normande herds in 6 French experimental farms. In the Holstein breed, we discriminated 4 BCS profiles based on combining BCS at calving (Low: around 2.6 points, or High: around 3.3 points) with BCS loss after calving (Moderate (M): ≤ 1.0 points, or Severe (S): > 1.0 points). The Low-M profile mostly included multiparous cows with higher milk yield and lower reproductive performance than cows in the 3 other profiles. Low-M cows that experienced abnormal ovarian activity had lower reproductive performance than their profile-mates. Moreover, 67% of Low-M cows kept the same profile at the following lactation. The High-S profile mostly included primiparous cows with lower milk yield and higher reproductive performance than cows in other profiles. In High-S cows, higher milk yields correlated to higher risk of failure to calf on first insemination. Moreover, 38% of High-S cows kept the same profile at the following lactation, and none changed to Low-M. The other 2 BCS profiles (Low-S and High-M) were intermediate in terms of milk yield and reproductive performance. In Normande, we discriminated 3 BCS profiles based on combining BCS at calving (Low: around 2.6 points, or High: around 3.5 points) with BCS loss after calving (Flat (F): flat with no loss, Moderate (M): around 0.5 points, or Severe (S): around 1.0 point). The Low-M and High-S profiles included cows with similar performance, even though High-S-profile cows showed better but not significantly different milk yield and reproduction performance. The High-F profile included cows that were more likely to experience abnormal ovarian activity and fail at first insemination than cows in other profiles. More than 50% of Normande cows with 2 successive lactations kept in the same BCS profile at the next lactation. Even though a low BCS at calving combined with severe BCS loss (more than 1 point) after calving was found to increase reproductive failure, there was no evidence of an optimal BCS profile for reproduction in dairy cows, and reproductive success or failure is multifactorial.

Sleep, Mood, and Nutrition Patterns of Postmenopausal Women Diagnosed with Major Depressive Disorder by Menopause Periods.

Menopausal women experience changes in ovarian activity and fluctuating hormone levels. The aim of this study was to detect ongoing sleep and nutritional problems in postmenopausal women. This study was conducted with 62 postmenopausal women who had been diagnosed with major depressive disorder, were aged 42-64, were not dieting for at least 1 month, and had been using antidepressants for at least 6 months. The Pittsburgh Sleep Quality Index and Depression Anxiety Stress Scale-42 were used. Anthropometric measurements were taken and body composition analysis was performed. The prevalence of obesity and overweight were 12.9% and 71%, respectively. Hot flashes, night sweats, and mood swing were more common in those who had been through menopause for <5 years. Also, the PSQI, Depression, and Stress scores of those women were higher. Women who had been menopausal for ≥5 years had a higher BMI, waist/hip ratio, and fat mass and better sleep quality and lower daytime dysfunction according to the PSQI. Energy and fat intake were higher, while protein, vitamin C, and calcium intakes were lower in women who had been menopausal for ≥5 years. It is crucial for healthcare professionals to review approaches for early and late menopausal periods and to individualize treatment options, especially in patients whose symptoms persist.

Dietary Aronia melanocarpa supplementation inhibit the oxidative stress caused by aging in ovary of late laying hens from 59 to 67 weeks of age

The ageing of laying hens’ ovaries in the late laying period leads to oxidative stress, which ultimately results in the decrease of laying performance. Thus, we investigated Aronia melanocarpa (AM) as a potent antioxidant to improve ovarian senescence in the late laying hens through antioxidant pathway, thereby improving egg production performance. A total of 480 Hy-line brown laying hens aged 59 wks (60–67 wks) were randomly divided into CON, LAM, MAM and HAM groups and fed 0, 1, 4 and 7% AM, respectively. The results showed that compared with the CON group, the MAM group could significantly improve the laying rate and reduce the feed egg ratio at 64–67 weeks of age (p < 0.05). The AM groups increased the eggshell thickness, while HAM group significantly reduced yolk colour (p < 0.05). The MAM and HAM groups significantly increased egg white weight (p < 0.05). Meanwhile, the ovarian index in MAM group was increased, and the proteins related to proliferation and anti-apoptosis of ovarian cells (PCNA and Bcl2/Bax) were enhanced in all AM groups (p < 0.05). Compared with the CON group, all AM groups could significantly reduce the malondialdehyde (MDA) content of ovary and yolk, and increase the activities of glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) in yolk. LAM and MAM groups significantly increased ovarian GSH-Px activity, but only the MAM group significantly increased ovarian T-SOD activity. The expression levels of Nuclear factor E2-related factor 2 (Nrf2), haem oxygenase 1 (HO-1) and glutathione peroxidase 1 (GPX1) genes and proteins were significantly increased in all AM groups, while the expression levels of kelch ECH – associated protein 1 (Keap1) genes and proteins were decreased. The expressions of NAD(P)H: quinone oxidoreductase 1 (NQO1) and superoxide dismutase 1 (SOD1) genes were significantly increased in all AM groups, but the expression of SOD1 protein was significantly increased in MAM group and NQO1 protein was significantly increased in HAM group. Antioxidant enzyme protein expressions in the ovary showed positive associations with antioxidant enzyme activity. Furthermore, the expression of Nrf2 protein in the ovary was inversely connected with Keap1 protein expression and favourably correlated with antioxidant protein expressions. In summary, dietary AM supplementation activates the Keap1/Nrf2 antioxidant pathway to inhibit the oxidative stress caused by ageing in ovary of late laying hens. Taken together, the results of the study suggest that the inclusion of 4% AM was able to better improve the laying performance and antioxidant capacity of laying hens in the late laying period.

Case Report: The Effect of Ajwa Dates on Calcium Levels in Perimenopausal Women

Perimenopause itself is defined as the permanent cessation of menstruation due to loss of ovarian activity confirmed by the onset of amenorrhea for 12 consecutive months in the absence of other pathological or physiological causes, occurring at the age of 42 years until menopause. In perimenopausal women, besides decreasing estrogen levels, there is also a decrease in calcium absorption which causes osteoporosis. Where dates contain iron, magnesium, calcium, phosphorus and potassium, but it also contains phytochemical compounds such as phytosterols and phytoestrogens. Phytoestrogens have the same structure and function as endogenous estrogens that serve to exert a major effect on women on bone remodeling and also control the timing of osteoclast apoptosis. Objective : To determine the relationship between giving ajwa dates to calcium levels in perimenopausal women. Method : Literature review with narrative review design. Results : Based on several journals that have been reviewed, it can be concluded that giving ajwa dates can inhibit the decrease in bone density in elderly women, where the active substances that play a role in this are calcium and phytoestrogens. Conclusion : Giving ajwa dates can inhibit the decrease in bone density in elderly women. Keywords : ajwa date (Phoenix dactylifera L), perimenopause, calcium.

Fertility Assessment after Ovarian Transposition in Children and Young Women Treated for a Malignant Tumor.

Ovarian transposition (OT) has been proposed as a protective measure against radiation-induced damage to ovarian function and fertility. Despite its historical use, limited research has focused on evaluating endocrine and exocrine ovarian function after OT performed in adolescents and young adults (AYAs) before or during puberty. The purpose of our study was to investigate the fertility, pubertal development, and ovarian function of women with a previous history of OT during childhood, adolescence or young adulthood. In an observational bicentric retrospective study, we included 32 young female cancer patients who underwent OT before the age of 26 between 1990 and 2015 at Lyon Léon Bérard Cancer Center or Nancy University Hospital. The mean age at the time of OT was 15.6 years with a cancer diagnosis at 15 ± 4.8 years. Among the 10 women attempting pregnancy post-treatment, 60% achieved successful pregnancies. After a mean follow-up of 9.6 ± 7 years, 74% (17 out of 23) of women recovered spontaneous menstrual cycles (seven out of eight evaluable women with OT before or during puberty). Notably, 35% of women who did not attempt pregnancy demonstrated adequate ovarian reserve. Ovarian reserve and function recovery were influenced by the specific chemotherapy received. Importantly, our findings suggest that OT's effectiveness on ovarian activity resumption does not significantly differ when performed before or during puberty compared to pubertal stages. This study contributes valuable insights into the long-term reproductive outcomes of young women undergoing OT, emphasizing its potential efficacy in preserving ovarian function and fertility across different developmental stages.

Fisetin mitigates letrozole-induced polycystic ovarian syndrome in rats: crosstalk of AMPK/PI3K/AKT-mediated-Nrf2 antioxidant defense mechanism and the inflammasome NLRP3/NF-κB P65/IL-1β signaling pathways.

Polycystic ovarian syndrome (PCOS) is a highly prevalent condition affecting reproductive-aged women, causing insulin resistance, hyperandrogenism, weight gain, and menstrual problems. The present study intended to investigate the potential role of fisetin (FT) in letrozole (LZ)-induced PCOS inadult female rats and the possible mechanism underlying its action. PCOS was induced by oral administration of LZ (1mg/kg) for 21days. Treated rats received FT (1.25 or 2.5mg/kg) orally once daily for 14 consecutive days. Following the experimental duration, blood samples and ovary tissues were isolated and preserved for biochemical and histopathological examinations. The results revealed that LZ-induced PCOS led to significant abnormalities in sex hormones and metabolic parameters. Additionally, it initiated an inflammatory cascade, evidenced by activation of the NF-κB p65/IL-1β and AMPK/PI3K/AKT pathways, alongside downregulation of Nrf2 ovarian gene expression and NLRP3 inflammasome activity, which enhanced the production of proinflammatory cytokines. FT demonstrated its beneficial impacts by restoring hormonal disturbance and reversing the imbalanced metabolic parameters. Moreover, FT increased the mRNA of ovarian Nrf2 levels and suppressed the up-regulated inflammatory IL-1β/NF-κB p65 signaling pathway, consequently alleviating the elevated levels of ovarian NLRP3. The histopathological examination also confirmed that FT has a beneficial effect in ameliorating PCOS, consistent with the aforementioned parameters. Finally, the present results demonstrated that FT ameliorates LZ-induced PCOS through the intricate interplay between the AMPK/PI3K/AKT-mediated Nrf2 antioxidant defense mechanism and the regulation of the inflammasome NLRP3/NF-κB p65/IL-1β signaling pathways.

Intravascular Laser Blood Irradiation (ILIB) Enhances Antioxidant Activity and Energy Metabolism in Aging Ovaries.

Ovarian aging is characterized by the accumulation of free radicals, leading to tissue damage and affecting reproductive health. Intravascular laser irradiation of blood (ILIB, using a low-energy He-Ne laser) is known for its efficacy in treating vascular-related diseases by reducing free radicals and inflammation. However, its impact on ovarian aging remains unexplored. This study aimed to investigate the effects of ILIB on oxidative stress and energy metabolism in aging ovaries. Genetic analysis was conducted on 75 infertile patients with aging ovaries, divided into ILIB-treated and control (CTRL) groups. Patients underwent two courses of laser treatment, and clinical parameters were evaluated. Cumulus cells were collected for the genetic analysis of oxeiptosis, glycolysis, and the tricarboxylic acid (TCA) cycle. The analysis of gene expression patterns revealed intriguing findings in ILIB-treated patients compared to the untreated group. Notably, ILIB treatment resulted in significant upregulation of oxeiptosis-related genes AIFM1 and NRF2, suggesting a potential protective effect against oxidative stress-induced cell death. Furthermore, ILIB treatment led to a downregulation of glycolysis-associated gene hexokinase 2 (HK2), indicating a shift away from anaerobic metabolism, along with an increase in PDHA levels, indicative of enhanced mitochondrial function. Consistent with these changes, ILIB-treated patients exhibited elevated expression of the key TCA cycle genes citrate synthase (CS), succinate dehydrogenase complex subunit A (SDHA), and fumarate hydratase (FH), signifying improved energy metabolism. The findings from this study underscore the potential of ILIB as a therapeutic strategy for mitigating ovarian aging. By targeting oxidative stress and enhancing energy metabolism, ILIB holds promise for preserving ovarian function and reproductive health in aging individuals. Further research is warranted to elucidate the underlying mechanisms and optimize the application of ILIB in clinical settings, with the ultimate goal of improving fertility outcomes in women experiencing age-related ovarian decline.

The β-arrestin1/endothelin axis bolsters ovarian fibroblast-dependent invadosome activity and cancer cell metastatic potential

Recruitment of fibroblasts to tumors and their activation into cancer-associated fibroblasts (CAFs) is a strategy used by tumor cells to direct extracellular matrix (ECM) remodeling, invasion, and metastasis, highlighting the need to investigate the molecular mechanisms driving CAF function. Endothelin-1 (ET-1) regulates the communication between cancer and stroma and facilitates the progression of serous ovarian cancer (SOC). By binding to Endothelin A (ETA) and B (ETB) receptors, ET-1 enables the recruitment of β-arrestin1 (β-arr1) and the formation of signaling complexes that coordinate tumor progression. However, how ET-1 receptors might “educate” human ovarian fibroblasts (HOFs) to produce altered ECM and promote metastasis remains to be elucidated. This study identifies ET-1 as a pivotal factor in the activation of CAFs capable of proteolytic ECM remodeling and the generation of heterotypic spheroids containing cancer cells with a propensity to metastasize. An autocrine/paracrine ET-1/ETA/BR/β-arr1 loop enhances HOF proliferation, upregulates CAF marker expression, secretes pro-inflammatory cytokines, and increases collagen contractility, and cell motility. Furthermore, ET-1 facilitates ECM remodeling by promoting the lytic activity of invadosome and activation of integrin β1. In addition, ET-1 signaling supports the formation of heterotypic HOF/SOC spheroids with enhanced ability to migrate through the mesothelial monolayer, and invade, representing metastatic units. The blockade of ETA/BR or β-arr1 silencing prevents CAF activation, invadosome function, mesothelial clearance, and the invasive ability of heterotypic spheroids. In vivo, therapeutic inhibition of ETA/BR using bosentan (BOS) significantly reduces the metastatic potential of combined HOFs/SOC cells, associated with enhanced apoptotic effects on tumor cells and stromal components. These findings support a model in which ET-1/β-arr1 reinforces tumor/stroma interaction through CAF activation and fosters the survival and metastatic properties of SOC cells, which could be counteracted by ETA/BR antagonists.

A Systemic Review of Pre-clinical Studies of Herbal Plants having Anti-Polycystic Ovarian Syndrome Activity: A PAN-India Study

Background: Polycystic ovarian syndrome (PCOS) is a prevalent endocrine and metabolic condition seen in premenopausal women. A number of pharmacological agents like metformin, oral contraceptives, and anti-androgen agents have been used to treat PCOS and its associated implications, but it has been associated with numerous negative effects like fatal and nonfatal lactic acidosis, weight gain, cardiovascular, and hepatic toxicity. Methodology: This study considered the Preferred Reporting Items for Systematic Review and Meta- Analysis (PRISMA) guideline where a vast number of article sources from different scientific databases like Google Scholar (https://scholar.google.com/), PubMed (https://pubmed.ncbi.nlm.nih.gov/), and Web of Science (https://mjl.clarivate.com/search-results) were screened out. Results: From a total of 154 studies, 24 studies were screened out following the inclusion and exclusion criteria, which reported 24 different plants that were used against PCOS preclinically. Details of various plants used, including their respective families, plant parts utilized, extraction methods employed, formulations developed, animal models of polycystic ovary syndrome (PCOS) utilized, and the geographical locations within India where the research has been conducted are all documented. Conclusion: This systematic review is the first in India to report the pre-clinical studies of medicinal plants against PCOS, and it provides a thorough summary of the application of medicinal plants for the management of PCOS. Nonetheless, molecular level investigations on the plants utilized, their bioactive phytoconstituent responsible for the anti-PCOS activity, the probable mechanism of action and the pathway through which they conserve their effect, the effective dose, and the potential adverse effects are the need of time.

Technological frontier in Uruguay's beef cattle production: An analysis of technical efficiency and its main drivers

AbstractGlobally, livestock grazing is a major land occupier and is often criticized for its environmental impact. To address these concerns, enhancing livestock productivity is crucial to mitigate these impacts, boost farm competitiveness, and increase food production. The understanding of production variability is key to this endeavor. This paper assesses the technical efficiency of cow‐calf beef production units in Uruguay, excluding dairy and feedlot operations. Using data from the 2016 National Livestock Survey and Wang's frontier model, we estimate a translog stochastic production frontier, incorporating bovine livestock units, land area, and equivalent working units as key inputs. This model also controls soil productivity, improved grazing areas, productive orientation (beef‐cattle systems or beef cattle‐ovine mixed systems), beef production system (cow‐calf, full cycle, and fattening operation), access to electric power, and agroecological regions. Furthermore, it explores factors affecting technical inefficiency, such as ownership (individual, corporate without contract, and corporate with contract) and land tenure types (owner, tenant, or other types), besides the presence of third‐party owned cattle, technical assistance (including veterinarian and agronomic), and specific beef cattle technologies (such as continuous bull presence, artificial insemination, ovarian activity diagnosis, pregnancy diagnosis, use of scales for classifying females, temporary and early weaning, and cows and heifers grazing together). Findings indicate an average technical efficiency of 71.3%. Notably, factors such as agronomic and veterinary assistance, artificial insemination, weaning practices, third‐party‐owned cattle presence, pregnancy diagnosis, land tenure, and ownership type significantly impact technical efficiency. Our findings suggest that within the specific segment of cow‐calf operations in Uruguay, there is a potential to enhance beef production by 40.2% through improvements in farm management. [EconLit Citations: Q12, D24].

Body Condition in Small Ruminants—Effects of Nutrition on the Hypothalamic–Pituitary–Gonad Axis and Ovarian Activity That Controls Reproduction

Nutritional status plays a vital role in regulating ovary activity. This regulation is mediated by the hypothalamus–pituitary–gonad axis and by effects exerted directly on the ovary. Therefore, to achieve the best reproductive performance, it is essential to know how the nutritional status affects the secretion of GnRH, gonadotrophins, and sex steroid hormones. Adequate body reserves and energy balance are critical for optimal reproductive performance in sheep and goats. However, over- or under-conditioned animals experience issues like extended anestrus, irregular ovarian cycles, and reduced conception. Body condition scoring allows for the evaluation of the relationships between adiposity, nutritional status, and fertility. Acute feed deficits briefly stimulate processes, but chronic restrictions suppress pulsatile LH release, disrupting ovarian function. The process of follicle development is a very complex one which involves intricate interactions between the pituitary gonadotrophins and metabolic hormones as well as between the locally produced factors by the ovarian somatic and germ cells including the IGF system and the TGF-β superfamily members. Genotype and nutrition are factors that have an impact on follicular development, and seasonal factors are also involved. This review will give a brief overview on how the body condition can be evaluated and the effects of nutrition on the hypothalamus–pituitary–gonad axis and ovarian activity, which are responsible for reproductive regulation. This paper presents a clear and reasonable summary of the pathway that runs from the nutritional status of small ruminants to ovarian activity through the hypothalamic–pituitary–gonadal axis. This review summarizes methods for body condition evaluation in small ruminants and evidence regarding acute versus prolonged nutritional impacts on the hypothalamic–pituitary–gonadal axis and ovarian activity controlling reproduction.

Determination of anti-Müllerian hormone levels in blood and urine in fertile cats

In recent years, blood anti-Müllerian hormone (AMH) levels have been investigated in female animals to diagnose many conditions, such as the presence of ovarian tissue, follicle reserve, and granulosa cell tumors. Since blood collection is an invasive method, diagnosis with a non-invasive method is important in terms of practicality and animal welfare. This study aimed to investigate the presence of AMH in cat urine and determine whether a correlation exists between blood and urine AMH levels. In addition, it was aimed at revealing whether there was a change in blood and urine AMH levels according to ovarian follicle distribution. Twenty-seven healthy, fertile female cats in the follicular phase were included. Following blood and urine sample collection, a routine ovariohysterectomy was performed. Histological analysis of the removed ovarian tissue was used to determine ovarian follicle types. While both AMH and estrogen levels were determined in blood samples, only AMH levels were investigated in urine samples. Blood AMH levels averaged 10.61 ± 0.75 ng/mL (range: 5–16 ng/mL), while urine AMH levels averaged 5.67 ± 0.91 ng/mL (range: 0.2–13 ng/mL). While urinary AMH level was <1 ng/mL in 7 cats, urinary AMH was >1 ng/mL in all remaining cats. While the study demonstrated AMH excretion in urine, no correlation was found between blood and urine AMH values. However, a significant positive correlation was observed between blood AMH levels and serum estrogen levels (P < 0.001). These findings suggest that urinary AMH may be a product of proteolytic degradation, potentially leading to inaccurate estimations of ovarian activity based solely on urine AMH levels.

PITPNA-AS1 Inhibits Cell Proliferation and Migration in Ovarian Cancer by Regulating the MIR-223-3p/RHOB Axis.

Ovarian cancer is a fatal gynecologic malignancy. Long non-coding RNA (lncRNA) has been verified to serve as key regulator in ovarian cancer tumorigenesis. The aim of the study was to study the functions and mechanism of lncRNA PITPNA-AS1 in ovarian cancer cellular process. Clinical ovarian cancer samples were collected and stored at an academic medical center. Cellular fractionation assays and fluorescence in situ hybridization were conducted to locate PITPNA-AS1 in OC cells. TUNEL staining, colony-forming assays, and Transwell assays were performed for evaluating cell apoptosis as well as proliferative and migratory abilities. Western blot was conducted for quantifying protein levels of epithelialmesenchymal transition markers. The binding relation between genes was verified by RNA pulldown, RNA immunoprecipitation, and luciferase reporter assays. Gene expression levels in ovarian cancer tissues and cells were subjected to RT-qPCR. PITPNA-AS1 level was downregulated in ovarian cancer samples and cells. PITPNA-AS1 overexpression contributed to the accelerated ovarian cancer cell apoptosis and inhibited cell migration, proliferation, and epithelial-mesenchymal transition process. In addition, PITPNA-AS1 interacted with miR-223-3p to regulate RHOB. RHOB knockdown partially counteracted the repressive impact of PITPNA-AS1 on ovarian cancer cell activities. PITPNA-AS1 inhibited ovarian cancer cellular behaviors by targeting miR-223-3p and regulating RHOB.

Oxytocin, prostaglandin F2α, and scopolamine for uterine involution of dairy cows.

The aim of the study was to compare the effect of three substances with ecbolic activity, Oxytocin, Prostaglandin F2α (PGF2α) and Scopolamine, on the uterine involution process in dairy cows and on the resumption of ovarian activity. Eighty bovine were randomly divided in four groups: GROUP C: 20 cows treated, within 24 h of calving, with 5 mL/head of saline solution; GROUP PG: 20 cows treated, within 24 h of calving, with 150 μg/head of d-cloprostenol; GROUP OX: 20 cows treated, within 24 h of calving, with 50 IU/head of oxytocin acetate; GROUP S: 20 cows treated, within 24 h of calving, with 40 mg/q Scopolamine Butylbromide. Each cow was subjected to blood samples to evaluate the Hydroxyproline (HYP) levels, at T0, within 24 h after calving, and T7, T14, T28, 7, 14, and 28 days after calving, respectively. At T14 and T28, an ultrasound examination was performed to measure the diameter of ex-pregnant horn. In all cows, the reproductive indices (days to first service and number of artificial insemination for conception) were evaluated. In all groups, the HYP concentrations have been rising from T0 to T28, with the maximum levels obtained at T28 in the groups PG and S. As regard the diameter of uterine horn, the comparison among the groups showed significant differences only at T28, with lower values in the group PG and S. In group S and PG, the days to first service were less than other groups. Treatment with Scopolamine and PGF2α resulted in better outcomes, evidenced clinically by more efficient uterine involution and faster ovarian recovery.

Molecular characterization and expression profile of the ALDH1A1 gene and its functions in yak luteal cells

The ALDH1A1 gene encodes a cytoplasmic member of the aldehyde dehydrogenase 1 family, which plays an important role in regulating animal reproductive performance, including estrus cycle and embryonic development. The aim of this study was to characterize ALDH1A1 activity in ovaries of 3–5 year-old yaks and to determine its effects on cell proliferation, apoptosis, and progesterone secretion in luteal cells (LCs). The coding sequence (CDS) of the ALDH1A1 gene was cloned by reverse transcription-PCR and immunohistochemical analysis was used to confirm localization of the ALDH1A1 protein in the ovary. To assess the activity of ALDH1A1 in regulating progesterone secretion, si-ALDH1A1 was transfected into LCs in vitro and progesterone levels in LC supernatants were measured by ELISA. The interference efficiency was assessed by real-time quantitative PCR (RT-qPCR) and immunofluorescence staining, and cell proliferation and apoptosis were evaluated by EdU and TUNEL staining, respectively. The cloned ALDH1A1 sequence contained 1462 bp, encoding 487 amino acids. Immunohistochemical analysis showed that ALDH1A1 protein expression, which was significantly higher in LCs, was mainly found in antral follicles and the corpus luteum (CL). The expression of ALDH1A1 mRNA in LCs was effectively inhibited by si-ALDH1A1transfection, and progesterone secretion was markedly decreased along with the significant down-regulation of progesterone pathway-related genes, STAR, CYP11A1, CYP19A1, CYP17A1, 3β-HSD, and HSD17B1. Knockdown of ALDH1A1 mRNA expression decreased cell proliferation and increased apoptosis in LCs. The mRNA expression of the proliferation-related genes, PCNA, CCND1, CCNB1 and CDC25A, was significantly down-regulated, while expression of the apoptosis-promoting CASP3 gene was significantly increased. In summary, we characterized the yak ALDH1A1 gene and revealed that ALDH1A1 knockdown promoted apoptosis, repressed cell proliferation, and decreased progesterone secretion by yak LCs, potentially by regulating the mRNA expression of genes related to proliferation, apoptosis, and progesterone synthesis and secretion.

Involvement of somatotrophic hormones in the postpartum regulation of ovarian activity in mares

Mares resume ovarian activity rapidly after foaling. Besides follicle-stimulating hormone (FSH) and luteinizing hormone (LH), the pituitary synthesizes prolactin and growth hormone which stimulate insulin-like growth factor (IGF) synthesis in the liver. We tested the hypothesis that follicular growth is initiated already antepartum, mares with early and delayed ovulation differ in IGF-1 release and that there is an additional IGF-1 synthesis in the placenta. Plasma concentrations of LH, FSH, IGF-1, IGF-2, activin and prolactin. IGF-1, IGF-2, prolactin and their receptors in placental tissues were analyzed at the mRNA and protein level. Follicular growth was determined from 15 days before to 15 days after foaling in 14 pregnancies. Mares ovulating within 15 days postpartum formed group OV (n=5) and mares not ovulating within 15 days group NOV (n=9). Before foaling, follicles with a diameter >1 cm were present in all mares and their number increased over time (p<0.05). Follicle growth after foaling was more pronounced in OV mares (day p<0.001, group p<0.05, day x group p<0.05) in parallel to an increase in LH concentration (p<0.001, day x group p<0.001) while FSH increased (p<0.001) similarly in both groups. Plasma concentrations of IGF-1 and prolactin peaked one day after foaling (p<0.001). The IGF-1 mRNA abundance was higher in the allantochorion but lower in the amnion of OV versus NOV mares (group p=0.01, localization x group p<0.01). The IGF-1 receptor mRNA was most abundant in the allantochorion (p<0.001) and IGF-1 protein was expressed in placental tissue without differences between groups. In conclusion, follicular growth in mares is initiated before foaling and placental IGF-1 may enhance resumption of ovulatory cycles.

ERβ Regulation of Indian Hedgehog Expression in the First Wave of Ovarian Follicles.

Increased activation of ovarian primordial follicles in Erβ knockout (ErβKO) rats becomes evident as early as postnatal day 8.5. To identify the ERβ-regulated genes that may control ovarian primordial follicle activation, we analyzed the transcriptome profiles of ErβKO rat ovaries collected on postnatal days 4.5, 6.5, and 8.5. Compared to wildtype ovaries, ErβKO ovaries displayed dramatic downregulation of Indian hedgehog (Ihh) expression. IHH-regulated genes, including Hhip, Gli1, and Ptch1, were also downregulated in ErβKO ovaries. This was associated with a downregulation of steroidogenic enzymes Cyp11a1, Cyp19a1, and Hsd17b1. The expression of Ihh remained very low in ErβKO ovaries despite the high levels of Gdf9 and Bmp15, which are known upregulators of Ihh expression in the granulosa cells of activated ovarian follicles. Strikingly, the downregulation of the Ihh gene in ErβKO ovaries began to disappear on postnatal day 16.5 and recovered on postnatal day 21.5. In rat ovaries, the first wave of primordial follicles is rapidly activated after their formation, whereas the second wave of primordial follicles remains dormant in the ovarian cortex and slowly starts activating after postnatal day 12.5. We localized the expression of Ihh mRNA in postnatal day 8.5 wildtype rat ovaries but not in the age-matched ErβKO ovaries. In postnatal day 21.5 ErβKO rat ovaries, we detected Ihh mRNA mainly in the activated follicles in the ovaries' peripheral regions. Our findings indicate that the expression of Ihh in the granulosa cells of the activated first wave of ovarian follicles depends on ERβ.

GnRH analogue triptorelin acetate induces ovulation in jennies during oestrus.

Most of the northern hemisphere donkey breeds are faced with the risk of extinction, thus donkey reproduction is considered an emerging branch of theriogenology, and the management of artificial insemination and induction of ovulation is a crucial point in setting up preservation protocols. For four consecutive cycles, six jennies' ovarian activity was routinely monitored; an ultrasound examination was performed daily from the evidence of a follicle diameter ≥27 mm until ovulation. Upon reaching a follicular diameter ≥32 ± 2 mm (Hour 0), oestrous jennies were treated alternatively with 0.1 mg triptorelin acetate, sc, (TRIP), 0.4 mg/sc of buserelin acetate (BUS) or saline, sc, (CTRL) and examined ultrasonographically at Hours 14, 24, 38, 42, 48, 62 and every 24 h until ovulation. Induction of ovulation was considered successful if ovulation occurred from 24 to 48 h after treatment. 11/12 cycles resulted in ovulation for TRIP and 12/12 for BUS and CTRL groups, respectively. Mean ± SD ovulation time after treatment was 37.3 ± 8.3, 47.1 ± 21.0 and 66.8 ± 25.9 h for BUS, TRIP and CTRL, respectively (p = .0032). Ovulation rates between h24 and h48 were 10/12 (83.3%) for both TRIP/BUS and 2/12 (16.7%) for CTRL, respectively (p = .003). Buserelin and triptorelin-treated jennies had a 25 times higher probability to ovulate between Hours 24 and 48 than controls (p = .003), while there were no jenny and cycle effects on the ovulatory rate. The results of this study show how triptorelin successfully induced ovulation in jennies, like other GnRH analogues previously evaluated.

Dynamics of serum exosome microRNA profile altered by chemically induced estropause and rescued by estrogen therapy in female mice.

The decline in the ovarian reserve leads to menopause and reduced serum estrogens. MicroRNAs are small non-coding RNAs, which can regulate gene expression and be secreted by cells and trafficked in serum via exosomes. Serum miRNAs regulate tissue function and disease development. Therefore, the aim of this study was to identify miRNA profiles in serum exosomes of mice induced to estropause and treated with 17β-estradiol (E2). Female mice were divided into three groups including control (CTL), injected with 4-Vinylcyclohexene diepoxide (VCD), and injected with VCD plus E2 (VCD + E2). Estropause was confirmed by acyclicity and a significant reduction in the number of ovarian follicles (p < 0.05). Body mass gain during estropause was higher in VCD and VCD + E2 compared to CTL females (p = 0.02). Sequencing of miRNAs was performed from exosomes extracted from serum, and 402 miRNAs were detected. Eight miRNAs were differentially regulated between CTL and VCD groups, seven miRNAs regulated between CTL and VCD + E2 groups, and ten miRNAs regulated between VCD and VCD + E2 groups. Only miR-200a-3p and miR-200b-3p were up-regulated in both serum exosomes and ovarian tissue in both VCD groups, suggesting that these exosomal miRNAs could be associated with ovarian activity. In the hepatic tissue, only miR-370-3p (p = 0.02) was up-regulated in the VCD + E2 group, as observed in serum. Our results suggest that VCD-induced estropause and E2 replacement have an impact on the profile of serum exosomal miRNAs. The miR-200 family was increased in serum exosomes and ovarian tissue and may be a candidate biomarker of ovarian function.