Corpus Albicans Research Articles

Exploring the distribution of polymorphism across diverse breeds Worldwide in the bovine NR5A2 gene and its correlation with number of mature follicles and corpus albicans

The Nuclear receptor subfamily 5 group A member 2 (NR5A2) gene plays a pivotal role in ovarian development, ovulation, and reproductive traits. There is a lack of studies on its impact on ovarian traits and reproductive traits in cattle. This study aimed to explore NR5A2 gene polymorphisms associations with reproductive traits and investigate the distribution of NR5A2 gene polymorphisms across diverse bovine breeds worldwide. We identified a novel 17-bp deletion within the NR5A2 gene specifically in Chinese Holstein cows (n = 1033) leading to the observation of two genotypes DD and ID. Subsequent association analysis revealed a significant correlation between the ‘ID’ genotype at this locus and a larger number of corpus albicans (p = 0.042) in diestrus, as well as a higher number of mature follicles (p = 0.038) in estrus. In addition, we also found that the distribution of this deletion exhibits strong regionality across different cattle breeds globally. These findings indicate that the 17-bp deletion mutation within the NR5A2 gene is significantly associated with an increased corpus luteum diameter and a greater number of mature follicles, suggesting its potential utility as a valuable DNA marker for enhancing cow fertility.

Luteal fibroblasts produce prostaglandins in response to IL1β in a MAPK-mediated manner

The corpus luteum is a temporary endocrine gland that is crucial for pregnancy, as it produces the progesterone needed to maintain optimal uterine conditions for implantation. In the absence of a conceptus, the corpus luteum becomes non-functional and undergoes rapid tissue remodeling to regress into a fibrotic corpus albicans. Early luteal regression is characterized by increased cytokine release. Because the role of fibroblasts in the bovine corpus luteum remains to be elucidated, the aim of this study was to elucidate the response of bovine luteal fibroblasts to inflammatory cytokines, tumor necrosis factor α (TNFα), and interleukin 1β (IL1β). Both cytokines induced canonical mitogen activated protein kinase (MAPK) signaling in luteal fibroblasts by phosphorylation of ERK1/2, p38 MAPK, and JNK. IL1β elevated expression and phosphorylation of cytosolic phospholipase A2 (cPLA2), an enzyme that mobilizes arachidonic acid for prostanoid synthesis. IL1β also elevated expression of prostaglandin-endoperoxide synthase 2 (PTGS2), another enzyme needed to synthesize prostanoids. IL1β increased PGF2α and PGE2 levels in the culture medium over 20-fold. Inhibition of MAPKs with small-molecule inhibitors abrogated the stimulatory effects of IL1β. IL1β also induced prostaglandin production in steroidogenic cells; however, there was no elevation in cPLA2. Therefore, actions of IL1β differ based on ovarian cell type. All together, we have identified luteal fibroblasts as potential inflammatory mediators during luteal regression.

Changes in surfactant protein A and D in ovine ovaries related to follicle development.

Surfactant protein A (SP-A) and Surfactant protein D (SP-D) glycoproteins play a crucial role in maintaining lung homeostasis and lung host defense. Interestingly, these proteins are also expressed in extra-pulmonary tissues, including the female genital tract. The ovarian tissue, where SP-A and SP-D expression increases with follicular development, may serve as the primary site of defense for this tissue. However, their functions in these tissues are not well understood and are currently an active area of research. Therefore, the objective of this study is to investigate the expression of SP-A and SP-D in the ovine ovary throughout the ovarian cycle using immunohistochemistry by semiquantitative intensity classification and Western blotting techniques. These findings revealed the presence of SP-A and SP-D in various compartments of the ovary, such as the follicular epithelium, granulosa cells, cumulus cells, theca cells, oocyte I, follicular fluid, and luteal cells of Graafian follicles, excluding the corpus albicans. SP-A and SP-D likely act as a first line of defense against potential pathogens that infiltrate the ovaries. Further investigation of the differential expression of SP-A and SP-D proteins in ovarian follicles will provide a basis for understanding their interactions with key proteins involved in oogenesis.

One-third of amenorrheic transmasculine people on testosterone ovulate

Transmasculine people usually reach amenorrhea within 6months of adequate testosterone treatment. It is often assumed that no ovulation occurs during amenorrhea. However, in this study, we report recent ovulatory activity in amenorrheic transmasculine people on testosterone therapy at gender-affirming oophorectomy. Histological signs of recent ovulatory activity, including the presence of ovulatory follicles, corpus luteum, and corpus albicans, are observed in 17 of 52 individuals (33%). This is not significantly correlated to the duration, testosterone serum levels, or type of testosterone used. These results suggest that amenorrhea does not equal anovulation in transmasculine people on adequate testosterone therapy, emphasizing the importance of contraception for people who engage in sexual activity that can result in pregnancy.

Background and common lesions in the female reproductive organs of giant anteaters (Myrmecophaga tridactyla).

The giant anteater (Myrmecophaga tridactyla) is a vulnerable species in South America and is considered endangered or near extinction in Central America. Therefore, studies describing the reproductive characteristics of this species are pivotal for its conservation. Thus, this study aimed to provide a morphological description of the female reproductive tissues of this species. We collected tissue samples from six female giant anteaters and performed gross, morphological, and histochemical analyses. Five adult subjects and one juvenile were included in the study. In the ovary, classifications were made according to the follicle and oocyte sizes: primordial, primary, secondary, early antral, or antral. Typical follicles with a single oocyte surrounded by a simple or stratified layer of cubic epithelium, atretic follicles, corpora lutea, corpora albicans, and ovarian cysts were also observed. No ovarian lesions were observed. By contrast, endometritis, metritis, mucometra, and endometrial cysts were identified in the uterus. Uterine alterations in these subjects were frequent and could affect reproduction.

Can blubber steroid hormone measurements reveal reproductive state in narwhals?

Abstract Hormone measurements from blubber samples have been used to assess reproduction in cetaceans and are a widely applicable technique, as blubber samples are often collected from necropsied individuals and biopsies are readily collected from live, free-swimming cetaceans. Many studies have assessed reproduction in cetaceans based on blubber hormone concentrations but few have validated their findings with individuals of known reproductive state. The objectives of this study were to use a unique dataset of paired female narwhal (Monodon monoceros) reproductive tracts and blubber samples to: (1) determine narwhal reproductive state based on ovarian analysis; (2) evaluate progesterone, estradiol, testosterone and corticosterone concentrations in paired blubber samples to validate the use of blubber hormone measurements as a technique to assess reproductive state in narwhals; and (3) determine narwhal reproductive rates using reproductive tract and hormone analyses. Female narwhals with complete reproductive tracts or known ages (n = 13) were categorized as: pregnant (fetus or placenta present; n = 5), active (at least one corpus luteum present; n = 2), resting (at least one corpus albicans present; n = 3) or immature (absence of corpora lutea and albicantia or age <8; n = 3), and eight individuals were classified as unknown due to incomplete reproductive tracts. Estradiol, testosterone, and corticosterone concentrations were not useful for assessing reproductive state; however, progesterone concentrations were higher in pregnant narwhals (432.66 ± 182.13 ng/g) than active (1.57 ± 0.42 ng/g), resting (1.52 ± 0.87 ng/g) and immature (1.44 ± 0.71 ng/g) individuals, validating the use of blubber progesterone concentrations in determining pregnancy in narwhals. Using a progesterone threshold for pregnancy, determined in this study, we were able to classify three individuals with incomplete reproductive tracts as pregnant and identify a potential impending pregnancy loss. The results from this study suggest that blubber progesterone concentrations are useful for assessing pregnancy and can help inform reproductive rates of narwhal populations.

Exploring the effects of methodological choices on the estimation and biological interpretation of life history parameters for harbour porpoises in Norway and beyond.

This study investigates effects of subtle methodological choices on the estimation and biological interpretation of age, growth and reproductive parameters for harbour porpoises. The core analyses are based on a focal Norwegian data set built on samples from 134 harbour porpoises caught incidentally in gillnet fisheries along the Norwegian coast during autumn 2016 and spring 2017. Two contrasting practices for interpretation of seasonal and ontogenetic characteristics of tooth growth layer formation resulted in significant age differences among spring samples of young porpoises and for older animals across seasons. In turn, these differences affected estimates of age at maturity and asymptotic lengths, respectively. We also found significant differences in male age at maturity between two well-documented maturity criteria and between mathematical estimators of age at maturity for both sexes. Two different criteria for corpus albicans classification furthermore resulted in different patterns of ovarian corpora accumulation, which may affect some estimates of fecundity rates and contaminant loads. Both corpora accumulation patterns were also found in reanalysed data from German and Greenlandic porpoises. Based on tabulated overviews of methodological choices made in previous harbour porpoise studies, we argue that several of the issues mentioned above have wider relevance and may affect the validity of meta-analyses as a tool for estimating harbour porpoise sensitivity to extrinsic pressures. Differences in cause of death (COD) composition between data sets can have a similar effect. We demonstrate this in a meta-analysis of published harbour porpoise pregnancy rates, showing significantly higher values for trauma-killed samples compared to samples comprising mixed COD categories. COD also affected the estimated impacts of three previously analysed extrinsic predictors as well as an added predictor for vessel noise levels. We discuss the potential contributions of methodological, biological and anthropogenic factors in shaping observed regional differences in estimates of harbour porpoise life history parameters.

Aged and induced-premature ovarian failure mouse models affect diestrus profile and ovarian features

Sex hormones exert a wide influence on several systems of the human body, especially in women, who undergo intense changes in the trans and postmenopausal periods. Different experimental models are used to mimic these conditions; however, the impact on hormonal profile may be different. This study aimed to analyze and compare vaginal cytology of different post-estropausal mice models, along with their microscopical ovarian features. Forty-six C57BL/6J female mice with the ages of 4, 6 and 18 months at the beginning of the experiment, weighing about 25–28 grams, constituted five groups: NC–(negative control) animals with no treatment, OVX-SHAM—sham ovariectomized, OVX–ovariectomized, VCD–medicated with 160 mg/kg/day of 4-vinylcyclohexene diepoxide via IP for 20 consecutive days, and Aged–senescent mice under physiological estropause. Euthanasia was performed at different periods for the removal of the ovaries, and after diestrus was confirmed by vaginal cytology for 10 consecutive days. For daily vaginal cytology, morphological and histomorphometric microscopic analyzes were performed. Aged mice presented significant increased neutrophils when compared to VCD group, as well as increased cornified epithelial cells when compared to OVX mice, and also increased nucleated epithelial cells when compared to VCD and OVX. NC and OVX-SHAM ovaries presented innumerous follicles at different stages of development, while VCD showed marked follicular atresia, depleted of primordial or developing follicles and a predominance of interstitial cells. The ovaries of aged mice were predominantly constituted by corpus luteum degenerated into corpus albicans, with rare antral follicles. All analyzed models led to different permanent diestrus profiles caused by each model, as indicated by ovarian features. This should be carefully considered when choosing a post-estropausal experimental model, in order to better correlate this challenging phase of female’s life with physiological/pathological conditions.

Immunoexpression of vascular endothelial growth factor and vWF-regulating angiogenesis in cyclic corpus luteum of Indian buffalo.

The present study was conducted to localize the immunoexpression of VEGF-A (Vascular Endothelial Growth Factor) and von Willebrand factor (vWF) in corpora lutea of healthy buffaloes (24) collected from local slaughterhouses. CL collected were categorized into early (stage I, 1-5 days, n = 6), mid (stage II, 6-11 days, n = 6), late luteal phase (stage III, 12 to 16 days, n = 6) and regressing phase (stage IV, 17 to 20 days, n = 6). The percent positive immunostaining for VEGF-A was significantly (p < 0.05) higher in mid-luteal phase than the other three stages of CL. However, it was higher in early luteal phase as well indicated intense angiogenesis in both early and mid-luteal phases. The number of capillary endothelium expressing vWF was significantly (p < 0.05) highest in mid-luteal phase among all the phases. However, in late luteal phase, the percent area positive for VEGF-A immunostaining was reduced but it was significantly (p < 0.05) higher than corpus albicans phase. Thus, in regressing phase or corpus albicans, it was lowest and reduced considerably. However, in late luteal phase, the number of capillaries with vWF immunoexpression reduced significantly (p < 0.05) but it was lowest in corpus albicans phase. Therefore, the immunotaining pattern for VEGF-A and vWF concluded that there was a spositive linear correlation between the two, that is, as the VEGF-A expression was increased, the number of vWF positive capillaries also increased and vice versa. The VEGF-A expressed by the luteal parenchyma in different stages of development and regression of corpus luteum was thus observed to be involved in promoting the angiogenesis and luteal cell proliferation as supported by vWF expressed by endothelium of proliferating capillaries in buffalo corpus luteum throughout the estrous cycle.

Mechanisms of angioregression of the corpus luteum.

The corpus luteum is a transient ovarian endocrine gland that produces the progesterone necessary for the establishment and maintenance of pregnancy. The formation and function of this gland involves angiogenesis, establishing the tissue with a robust blood flow and vast microvasculature required to support production of progesterone. Every steroidogenic cell within the corpus luteum is in direct contact with a capillary, and disruption of angiogenesis impairs luteal development and function. At the end of a reproductive cycle, the corpus luteum ceases progesterone production and undergoes rapid structural regression into a nonfunctional corpus albicans in a process initiated and exacerbated by the luteolysin prostaglandin F2α (PGF2α). Structural regression is accompanied by complete regression of the luteal microvasculature in which endothelial cells die and are sloughed off into capillaries and lymphatic vessels. During luteal regression, changes in nitric oxide transiently increase blood flow, followed by a reduction in blood flow and progesterone secretion. Early luteal regression is marked by an increased production of cytokines and chemokines and influx of immune cells. Microvascular endothelial cells are sensitive to released factors during luteolysis, including thrombospondin, endothelin, and cytokines like tumor necrosis factor alpha (TNF) and transforming growth factor β 1 (TGFB1). Although PGF2α is known to be a vasoconstrictor, endothelial cells do not express receptors for PGF2α, therefore it is believed that the angioregression occurring during luteolysis is mediated by factors downstream of PGF2α signaling. Yet, the exact mechanisms responsible for angioregression in the corpus luteum remain unknown. This review describes the current knowledge on angioregression of the corpus luteum and the roles of vasoactive factors released during luteolysis on luteal vasculature and endothelial cells of the microvasculature.

Response to hormonal treatment and conception rates of Sahiwal cows subjected to fixed time artificial insemination in pastoral dairy systems.

This study aimed at determining factors influencing response of Sahiwal cows/heifers to fixed time artificial insemination protocol in pastoral systems in Kenya. Available cows/heifers were inspected for conformity to Sahiwal breed characteristics, parity, body condition score, and subsequently rectal palpation to determine pregnancy status, ovarian structures, and estimated ovarian diameter. Consequently, these animals were injected with 100µg of gonadotrophin-releasing hormone. On days 7 and 9, only responsive cows/heifers were injected with 500µg of cloprostenol and 100µg of gonadorelin Acetate, respectively. On day 10, animals were inseminated and separated from bulls for 45days and pregnancy diagnosis done after 90days. Analysis of variance was performed to determine the effects of production system, parity, and ovarian structures on ovary diameters pre- and post-hormonal treatment. Logistic regression was used fitting a logit function to account for the binomial distribution of conception. Overall, 56.2%, 23.1%, and 20.7% of the animals had follicles (F), corpus luteum (CL), and corpus albicans (CA), respectively, at day 0, and 16.6%, 68.6%, and 14.8%, respectively, at day 7. Human and environmental factors had no influence on conception. Among the animal factors, only the ovarian structures at day 7 had a significant effect on conception. Ovaries with CL at this time were about 6 times significantly more likely to conceive than those with F. For higher conception rates, animals with ovaries with CL should be recruited into the FTAI program as they are significantly more likely to conceive than those with other ovarian structures.

Giant Ovarian Corpus Luteum Cysts Associated with Natural Pregnancy in Woman with Polycystic Ovary Syndrome

A 29-year-old nulliparous woman with polycystic ovary syndrome and no history of ovarian cyst was admitted to obstetric clinic for prenatal screening at 20-week spontaneous gestation of twins. An ultrasound examination showed multicompartmental cysts whose diameters were 20 cm and 15 cm on the left and right ovaries, respectively. A cancer antigen 125 (CA-125) test showed an elevated level of 209.0 U/mL, and the level of alpha fetoprotein (AFP) was 170.0 ng/mL. A diagnosis of physiological cysts was considered. Then, the woman was conservatively managed, and cesarean delivery was scheduled. The size of the cysts increased gradually with the week of gestation, and the diameters got their maximum at 30 cm and 20 cm for each side until 30 weeks (Fig. 1). Unfortunately, she presented spontaneous premature labor then. No adverse maternal or neonatal outcomes were reported. At her 2-month check after delivery, the CA-125 and alpha fetoprotein decreased to normal range. Nevertheless, 6 months after delivery, the sizes of the ovaries were still 20 cm and 10 cm in diameter at the left and right sides, respectively, accompanied by intermittent abdominal pain. Subsequently, she underwent single-port laparoscopic oophorocystectomy (Fig. 2). Pathological analysis revealed bilateral corpus albicans (Fig. 3). The patient recovered smoothly. We present a case of physiological ovarian cysts of unusual size during pregnancy with elevated CA-125, resulting in premature delivery. Expectant management may be considered for giant lutein cyst in pregnancy. However, if there is any suspicion of a malignancy or complications caused by cysts, ovarian cystectomy during pregnancy should be performed.Fig. 2Laparoscopy revealed the huge polycystic ovaries. (A) The pelvis was filled with giant cysts. (B) The left ovary (yellow arrow) was squeezed by the cysts.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig. 3The pathologic photomicrograph revealed corpus albicans.View Large Image Figure ViewerDownload Hi-res image Download (PPT) The authors thank Lei Pan and Kui Li, who aided in data collection and image production.

Determination of puberty in gilts: contrast of diagnostic methods

BackgroundEarly onset of a gilt´s puberty is needed for adequate economic performance in farms, because it indicates her reproductive performance and longevity. Therefore, an effective diagnosis is needed. Our purpose was to compare different procedures (external characteristics, blood progesterone analysis and ultrasonography diagnosis) to detect puberty in 70 gilts (Topigs TN70; 240 days old) on farm conditions. Postmortem examination was the standard reference. Multiple logistic regression analysis was used to identify which combination of independent variables (predictors) best predicts the status of gilts.ResultsPuberty (46/70 gilts; 65.71%) was characterized by the presence of follicles larger than 6 mm, corpus albicans, corpus rubrum, and corpus luteum (postmortem examination). Vaginal length, body condition, backfat, carcass weight and progesterone blood concentration were significantly higher in pubertal than prepubertal gilts (P < 0.05). Two types of ultrasonography equipment (DELTA and W3) were compared and performed by the same senior technician (V1). The results obtained by two technicians with different levels of experience (V1 and V2, a junior technician) using W3 were also compared. Ultrasonography provided better results than other diagnostic techniques, although the effectiveness of the ultrasonography changed with technological improvements and with increased expertise of technicians. The most accurate results were found by V1/DELTA (Nagelkerke´s R2 = 0.846; Sensitivity = 0.956; Specificity = 0.958; Positive predictive value = 0.978; Negative predictive value = 0.920; Area under ROC curve = 0.957). Results using the W3 equipment could be improved when used in conjunction with vaginal length (V1; Nagelkerke´s R2 = 0.834; Sensitivity = 0.933; Specificity = 0.958; Positive predictive value = 0.977; Negative predictive value = 0.885; Area under ROC curve = 0.972) or progesterone concentration (V2; Nagelkerke´s R2 = 0.780; Sensitivity = 0.955; Specificity = 0.826; Positive predictive value = 0.915; Negative predictive value = 0.905; Area under ROC curve = 0.970).ConclusionsUltrasonography provided better results than other diagnostic techniques. The effectiveness of the ultrasonography changes with technological improvements and with increased expertise of technicians. Results using the W3 equipment could be improved when used along with vaginal length (V1) or progesterone concentration (V2). Accuracy parameters are a guide to choose puberty diagnosis, but the farms must also evaluate effect on gilts, ease and cost of administration.

Expression of dihydrotestosterone synthases and androgen receptor in sheep oviduct ampulla and its regulation by estradiol and progesterone

Oviduct ampulla plays an important role in steroid hormone-regulated sperm-oocyte binding in female animals. Although studies have shown that androgen receptor are expressed in many species oviduct, the interaction among androgen receptor (AR), estradiol (E2) and progesterone (P4) in the sheep oviduct have rarely been reported. In this study, we evaluated the localization of two isoforms of dihydrotestosterone (DHT) sythetase enzymes 5α-reductase (5α‐red1, 5α‐red2) and AR in sheep oviduct ampulla by immunohistochemistry and immunofluorescence. Results showed that they were all distributed in oviduct epithelium layer. In epithelial cells, 5α‐red1, 5α‐red2 were expressed in cytoplast and nuclear, but AR were stained in nuclear. We also investigated their expression pattern in the sheep oviduct ampulla at different development stages of follicles (Large follicles stage; hemorrhagium, luteum and albicans of corpus stage) by molecular experiments. We found that 5α‐red1, 5α‐red2 and AR mRNA abundance and protein were expressed highest in corpus albicans stage and lowest in corpus hemorrhagium stage. In vitro, when sheep oviduct ampulla epithelial cells (SOAECs) were cultured and treated with different concentrations of E2/P4 (10−9–10−6 M), we found that E2 inhibited the expression of AR mRNA and protein, while P4 promoted this expression. In addition, when the SOAECs were treated with E2 (10−8 M) and/or its non-selective inhibitor ICI182780 (10−7 M) as well as with P4 (10−6 M) and/or its non-specific inhibitor RU486 (10−5 M), we found that E2 and P4 inhibited and promoted the expression of AR mRNA and proteins, respectively, via their nuclear receptor pathways. This study provides a basic insight for the further research of oviduct epithelium physiological function closely related to androgen.

Экспрессия miR-181a и miR-146a при преждевременной овариальной недостаточности аутоиммунной этиологии

Objective. To study the expression of miR-181a and miR-146a in the blood serum obtained from an experimental animal model of autoimmune oophoritis on different days of the experiment to assess their role in primary ovarian insufficiency (POI) of autoimmune etiology. Materials and methods. Experimental animals were mature female outbred rats. The study group (n = 25) included the experimental animal model of autoimmune oophoritis, the control group (n = 20) – intact animals. Morphometry of ovarian samples with estimating the average number of follicles, corpora lutea and atretic corpora lutea was performed. Enzyme immunoassay was used to study the concentration levels of serum anti-Mullerian Hormone (AMH) and anti-ovarian antibodies (AOA). MicroRNAs were isolated from serum according to a standard protocol; a reverse transcription and real-time polymerase chain reaction were performed using specific primers, and the relative expression of miR-181a and miR-146a was calculated by the Pfaffle method. Results. The expression of miR-146a on different days of the experiment remained statistically insignificant, including when compared to the control group. The expression of miR-181a in the blood serum of experimental animals on day 15 was statistically significantly higher than similar values on days 5, 10, 30 and 60 and in the control group. Conclusion. MiR-181a showed high diagnostic value in POI of autoimmune etiology in the experiment performed and can serve as an early marker in the diagnosis of ovarian insufficiency. The role of miR-146a as a marker of POI of autoimmune etiology has not been experimentally confirmed. Key words: autoimmune oophoritis, microRNA, experiment, premature ovarian insufficiency

Expression profile of the Kisspeptin/Kiss1r system and angiogenic and immunological mediators in the ovary of cyclic and pregnant cats

The Kisspeptin/Kiss1r system has been studied in mammalian ovaries. However, there are still no studies on the modulation of this system and its relationship with angiogenic and immunological mediators in the ovary of domestic cats, especially during pregnancy. We evaluated the expression of Kisspeptin/Kiss1r and angiogenic and immunological mediators during folliculogenesis, luteogenesis and luteal regression of cyclic and pregnant cats. The ovary exhibited moderate to intense expression for Kiss1, VEGF, Flk-1, INFγ and MIF in oocytes and the follicular wall, while Kiss1r expression was low in granulosa cells. In these cells, there was also a greater expression of Kiss1, INFγ and MIF, mainly in secondary follicles, while tertiary and preovulatory follicles exhibited greater expression of VEGF and Flk-1 in this layer. In luteogenesis, Kiss1 immunostaining was higher in mature corpora lutea (MCL) of pregnant cats compared to vacuolated CL (VCL) and corpus albicans (CA). Pregnancy also increased the luteal gene expression of Kiss1 as well as Kiss1, Kiss1r, Flk-1, and MIF immunostaining in MCL, while reduced the area of VEGF expression in VCL and luteal mRNA expression of Mif when compared to non-pregnant animals. In addition, positive gene correlation between Kiss1r and Mif was observed in the CL. Kiss1, Kiss1r, Vegf and Mif expression were lower in the CA of cats in anestrus. These findings reveal that the expression of Kisspeptin/Kiss1r and angiogenic and immunological mediators, in the ovary of domestic cats, depend on the follicular and luteal stage, and the luteal expression of these mediators is influenced by pregnancy.

Effects of shikonin on the development of ovarian follicles and female germline stem cells

ObjectiveTo investigate the effects and potential mechanism of action of shikonin (SHK) on the development of ovarian follicles and female germline stem cells (FGSCs).MethodsFemale Kunming adult mice were administered SHK (0, 20 and 50 mg/kg) by oral gavage. Cultures of FGSCs were treated with SHK 32 μmol/l for 24 h. The ovarian index in mouse ovaries was calculated. Numbers of primordial, primary and atretic follicles were counted. Germline stem cell markers and apoptosis were examined. Levels of glutathione (GSH), superoxide dismutase (SOD) and reactive oxygen species (ROS) were measured.ResultsBoth doses of SHK significantly decreased the ovarian index, the numbers of primordial follicles, primary follicles and antral follicles in mice. SHK significantly increased the numbers of atretic follicles and atretic corpora lutea. SHK promoted apoptosis in vivo and in vitro. SHK significantly decreased the levels of the germline stem cell markers. SHK significantly lowered GSH levels and the activity of SOD in the peripheral blood from mice, whereas SHK significantly elevated cellular ROS content in FGSCs.ConclusionsThese current results suggested that follicular development and FGSCs were suppressed by SHK through the induction of apoptosis and oxidative stress might be involved in this pathological process.

HISTOLOGY AND HISTOMORPHOMETRY OF GAYO MARE OVARIES

This study aims to determine the histology, the number and diameter of follicles, and the corpus albican of the Gayo mare ovary. The micro technical process was applied to 3 pairs of ovarian samples for further hematoxylin-eosin (HE) staining. Observation of ovarian structure was carried out microscopically and the data were analyzed statistically. The results showed that the Gayo mare ovary consists of the medulla on the outside and the cortex on the inside. Medulla consists of small follicles, blood vessels, nerves, and connective tissue. Meanwhile, the cortex consists of de Graff's follicle, corpus luteum, corpus albican, and ovulatory fossa. Follicles are composed of oocytes, granulosa cells, internal and external theca cells, oophorous cumulus, follicular antrum, and follicular fluid. Atretic follicles contain lutein cells that have been damaged. The corpus luteum is composed of granulosa lutein tissue, internal theca and external. The corpus albican is composed of scar tissue and lutein cells. The measurement results showed that the primordial follicle diameter was 26.60±2.37 µm, primary follicle 54.33±6.70 µm, secondary follicle 119.32±25.55 µm, tertiary follicle 250.86±49.46 µm, atretic follicle 49.03±45.47 µm, the corpus albican 511.10±132.41 µm. Thus, it can be concluded that the Gayo mare ovary has a histological structure that is not different from the ovary of other mares. Follicular growth occurs in the medulla of the ovary, and de Graff's follicles are present in the cortex of the ovaries.

Polymorphic variants of bovine ADCY5 gene identified in GWAS analysis were significantly associated with ovarian morphological related traits

The reproductive performance (e.g. fertility) of dairy cows, which declined over past few decades due to the intense and intensive selection, needs to be improved. Previous genome-wide association study (GWAS) of female Holstein screened the Adenylate cyclase 5 (ADCY5) as the candidate gene for cow fertility. As a member of the adenylyl cyclases family, adenylate cyclase 5 (ADCY5) is famous for regulating extrapyramidal motor system related various neuropsychiatric diseases, and its genetic variant is reported to associate with lower birth and placenta weight which leads to asymmetric fetal growth restriction. It was hypothesized that ADCY5 may affect the fertility of cows by regulating the processes of ovarian development. Herein, genomic DNA from 768 ovaries samples of healthy unrelated Holstein cow were used to screen potential insertion/deletion (indel) mutations using eight pairs of primers, and we found three novel polymorphic indel variants, namely, rs385624978 (P3-D11-bp), rs433028962 (P5-I19-bp) and rs382393457 (P8-D19-bp). The minor allelic frequencies (MAF) of P3-D11-bp, P5-I19-bp and P8-D19-bp loci were 0.188, 0.365 and 0.06, respectively, and there were 7 different haplotypes. Additionally, linkage disequilibrium analysis demonstrated no linkage among them. Importantly, P3-D11-bp locus was significantly related to both ovarian width (P = 1.0E-6) and corpus luteum diameter (P = 0.015); P5-I19-bp locus had a significant relation with corpus albicans diameter (P = 0.030) and ovaries with mutational homozygous genotype produced a superior corpus albicans diameter than those with other genotypes. Briefly, three novel indel mutations of bovine ADCY5 gene were identified and two of them were uncovered to be significantly correlated with ovarian phenotypic traits or corpus luteum or albicans traits. These findings contributed to the application of molecular marker-assisted selection (MAS) in improving female fertility in cattle, which could accelerate the development of the cattle industry.

Effect of Point Mutation in the Growth Differentiation Factor 9 Gene of Oocytes on the Sterility and Fertility of Awassi Sheep.

Growth differentiation factor 9 (GDF9) plays a critical role in ovarian follicular development and ovulation rate. The present study aimed to investigate the correlation between the single-nucleotide polymorphism (SNP) of the GDF9 gene and reproductive performance variables, such as fertility and sterility in Awassi sheep. Forty pairs of ovaries from a total of 40 slaughtered Iraqi Awassi ewes were used in this study. Twenty of the ovaries were collected from sterile ewes and the other 20 ovaries were taken from fertile ewes for genomic DNA extraction, polymerase chain reaction, and sequencing to detect GDF9 gene polymorphism. Follicles and oocytes of all the 40 ovaries were evaluated and compared with the results of genotyping. Furthermore, histopathological and microscopic evaluations were performed for 40 ovarian tissues of the two groups. The sequence analysis revealed that exon I had three SNPs, including T(114)C, G(129)R, and G(199)A. The first two SNPs were silent mutations and the last mutation was missense responsible for the substitution of glutamic acid with lysine at position 67. The current study showed a significant increase (P&amp;le;0.01) in GG, AA, CC, GA, and GG genotypes at G(129)R, G(199)A, T(114)C, G(129)R, and G(199)A loci, respectively. Moreover, the TT genotype in locus T(114)C was recorded to significantly augment (P&amp;le;0.05) in the fertile ewes. Mutant GA genotype of the G(129)R locus led to a significant (P&amp;le;0.05) increase in the percentage of follicles (4-8 mm) and oocytes number, compared to wild GG. On the other hand, a significant decrease was recorded in the mutant AA genotype in G(199)A, compared to wild GG. Differences between CC and TT genotypes at T(114)C locus were not significant. Histopathological examination revealed hypoplasia in the ovarian tissue of sterile ewes accompanied by fibrous connective tissue invasion and follicles degeneration. However, in the fertile ewes, the ovarian tissues were normal with the presence of numerous corpus albicans and degenerative corpus luteum. According to the findings of this study, the homozygote mutation in fertile ewes minimized the number of follicles and oocytes leading to sterility, while the heterozygote mutation was reported in the fertile Awassi ewes.