Luteal Research Articles

Transcriptomic analysis of luteal tissue supports the earlier onset of luteolysis in heifers with diminished ovarian reserve.

Context Antral follicle count (AFC) is reflective of ovarian reserve and linked to reproductive performance in mammalian females. We previously demonstrated earlier upregulation of endometrial oxytocin receptor transcription in heifers with low AFC, indicating an earlier onset of luteolysis. Aims We aimed to support the earlier onset of luteolysis in non-pregnant heifers with a low number of antral ovarian follicles (Open Low AFC) and hypothesized a reduced abundance of luteinizing hormone/choriogonadotropin receptor (LH/CG-R ) and increased abundance of thrombospondin-1 (THBS1 ) in luteal tissue of those heifers. We further investigated if classical interferon stimulated genes (ISGs) are already upregulated in response to conceptus-derived interferon tau. Methods For 4years, 10 heifers with the highest (High AFC) and 10 heifers with the lowest AFC (Low AFC) of the population (n =120/year) were selected, synchronized and inseminated. On day 15 or 16 after insemination, reproductive tracts were collected, and pregnancy status was determined. Corpora lutea were isolated, weighed and underwent transcriptomic analysis. Key results Transcript abundance of LH/CG-R was lower in Open Low AFC heifers compared to all other subgroups. Transcript abundance of THBS1 was upregulated in Open vs Pregnant heifers and showed greater abundance in Low vs High AFC heifers. Luteal weights and abundance of ISGs did not differ between heifers of differing AFC groups or pregnancy status. Conclusions The patterns of luteal LH /CG-R and THBS1 abundance support the concept of earlier onset of luteolysis in Open heifers with diminished AFC. Implications This may result in a shorter window for maternal recognition of pregnancy and contribute to inferior reproductive performance.

The histological investigation of the effects of electromagnetic radiation on rat ovaries.

People are now exposed to higher levels of electromagnetic radiation (EMR) due to the widespread use of mobile phones in recent years. The possible effects of this exposure on human health are related to EMR. It has been suggested that exposure to EMR has serious effects on reproduction. The study aimed to investigate the impact of exposure to EMR (4.5GB; 2600MHz) emitted by mobile phones on rat ovaries. 18 adult female Wistar albino rats were used in the study, and the animals were divided into three groups (n = 6): control, stand-by, and dialing. For 8weeks, the experimental groups were subjected to 4.5GB EMR at 2600MHz while on standby and making 10-min calls every hour. The rats in the control group received no exposure. Hematoxylin-eosin (H&E) staining of ovarian tissues was performed for histomorphological examinations. Additionally, immunoexpression of autophagy-related protein Beclin-1, apoptosis marker Caspase-3, ovarian reserve marker FSH, and oxidative stress marker iNOS were investigated in the rat ovaries. Microscopic examinations showed follicular degeneration in the ovaries of the rats in the stand-by and dialing groups. The immunoexpression of Beclin-1, Caspase-3, FSH, and iNOS was detected in granulosa cells and the corpus luteum in ovarian tissues obtained from the two EMR-exposed groups. There was a significant increase in the immunoexpression of Beclin-1 and Caspase-3 in the dialing group compared to the other two groups. Additionally, the iNOS and FSH expressions were increased in both EMR exposure groups compared to the control. Our results suggest that EMR exposure harms the ovaries, and autophagy and apoptosis are involved in this process.

EFFECTS OF INJECTABLE MINERAL AND VITAMIN SUPPLEMENTATION ON THE FERTILITY OF DAIRY COWS IN FTAI PROTOCOL

The study aimed to assess the impact of injectable mineral and vitamin supplementation, specifically Kit Adaptador®MIN and Adaptador®VIT (Biogénesis Bagó, Brazil), administered at the initiation of a fixed-time artificial insemination (FTAI) protocol, on the reproductive performance and fertility of Holstein cows. One hundred animals were divided into a non-supplemented control group (CG, n=50) and a treated group (TG, n=50). The TG received mineral and vitamin supplementation (1 mL/100kg live weight, IM) on the first day of the FTAI protocol (D0), along with intravaginal progesterone (P4) implantation and estradiol benzoate administration. Blood samples were collected at D0 and D17 to assess superoxide dismutase (SOD) enzyme activity and P4 levels. Ovarian and uterine ultrasound evaluations were conducted at D0 and D17, with follow-up exams performed 30 days post-FTAI for pregnancy diagnosis. Results showed a larger corpus luteum diameter in supplemented cows (CG=2.14±0.11 cm vs TG=2.64±0.11 cm, P=0.0037), along with increased SOD activity (CG=22% vs TG=25.36%, P=0.045). Moreover, there was a significant difference in pregnancy rates between groups (CG=18.18% vs TG=39.13%, P=0.0029). Injectable mineral and vitamin supplementation at the start of the FTAI protocol may impact post-ovulation corpus luteum development, enhance pregnancy rates, and stimulate essential endogenous antioxidant mechanisms in Holstein cows.

Impact of corpus luteum number on maternal pregnancy and birth outcomes: the Rotterdam Periconception Cohort.

To investigate associations between assisted reproductive technology (ART) -induced alterations in corpus luteum (CL) number during implantation, and maternal pregnancy and birth outcomes. Pregnancies conceived through ART are associated with increased risks of adverse obstetric and perinatal outcomes, with discrepancies in outcomes between different ART treatment protocols. We hypothesize this is due to the periconceptional hormonal environment regulated by the CL. ART protocols affect CL quantity at conception resulting in CL absence or a supraphysiological number of CL. This study is embedded in the Rotterdam Periconceptional Cohort, an ongoing tertiary center prospective cohort at the Erasmus University Medical Center in Rotterdam, the Netherlands. Women with a singleton pregnancy with data on CL. The number of CL, based on mode of conception: 0 CL (artificial cycle frozen embryo transfer (AC-FET), n=72); >1 CL (ovarian stimulated fresh embryo transfer, n=462) and 1 CL (natural cycle frozen embryo transfer (NC-FET) and natural conceptions, n=1327). Hypertensive disorders of pregnancy, gestational diabetes, gestational age at birth and birthweight, derived from medical records. We included 1,861 pregnancies. The results were adjusted for maternal age, maternal body mass index, nulliparity and obstetric history. In comparison with natural conceptions (1 CL), a pregnancy with CL absence (0 CL) was associated with a higher risk of gestational diabetes (aOR 2.59 [95% CI 1.31;5.15]), and a higher risk of preeclampsia, albeit non-significantly (aOR 2.02 [95% CI 0.91;4.51]). In comparison with pregnancies with >1 CL the risk of preeclampsia was significantly lower (aOR 0.36 [95% CI 0.18;0.72]). Post hoc analyses revealed that in male neonates, >1 CL was associated with a lower birthweight percentile (aβ -6.18 [95% CI -11.16;-1.20]). In contrast, female neonates showed no association with >1 CL, whereas CL absence was associated with a higher birthweight percentile (aβ 12.93 [95% CI 2.52;23.34]). Risks of hypertensive disorders of pregnancy, gestational diabetes and relative birthweight differ between CL groups. These findings support the hypothesis that an aberrant number of CL impacts maternal pregnancy and or birth outcomes. Additional studies need to be conducted to investigate causes and underlying pathophysiology.

APN/AdipoRon regulates luteal steroidogenesis through AMPK/EZH2/H3K27me3 in goats.

AMPK plays a crucial role in cellular energy metabolism and is involved in the regulation of luteal steroidogenesis by APN and its analog AdipoRon. To further explore the regulatory mechanism of AMPK in goat luteal steroidogenesis mediated by APN, cyclic and pregnant CL were utilized to assess the localization and expression of AMPK, EZH2, H3K27me3 and H3K27ac by WB and mIHC, and the interaction between AMPK and EZH2 by Co-IP. Then, isolated luteal cells were treated with APN/AdipoRon to evaluate the expression levels of AMPK, EZH2, H3K27me3 and H3K27ac. Results showed that AMPK and EZH2 were co-localized to the cytoplasm of luteal cells, and interacted as detected by Co-IP. H3K27me3 and H3K27ac were localized to the nucleus of goat luteal cells. H3K27me3 expression in late CL was significantly higher than that in early and middle CL, while the expressions of AMPK, H3K27ac and EZH2 in middle CL were significantly higher than those in early and late CL. Notably, all these proteins were expressed at similar levels between pregnancy and middle cycle, with the exception of EZH2. Following incubation with AdipoRon (25μM) and APN (1μg/mL) for 24h, the expressions of AMPK and H3K27ac decreased, while H3K27me3 increased in luteal cells. Compound C (AMPK activity inhibitor) reversed the AdipoRon - induced decrease in EZH2 expression and the increase in H3K27me3 expression. The increased H3K27me3 expression and decreased steroidogenic protein (CYP11A1 and HSD3B) expression after GSK126 (EZH2 inhibitor) treatment were consistent with the effects seen after AdipoRon treatment. In conclusion, APN/AdipoRon inhibits luteal steroidogenesis by inhibiting the interaction between AMPK and EZH2, thereby promoting H3K27me3 expression.

Hypoxia-Inducible Factor inhibition affects luteal function with no effect on fertility in mice.

Hypoxia-inducible factors (HIFs) are transcription factors responsible for sensing low oxygen levels and, in response, inducing the transcription of numerous genes. One of the main processes stimulated by HIFs is the formation of new vessels to increase oxygen supply to the tissue. Formation of the corpus luteum strongly depends on the vasculature and an active angiogenesis occurs during luteinization. In this study, we aimed to analyze the role of HIF in corpora lutea early formation and function, and in female fertility. To this aim, we superovulated mice by eCG and hCG, and administered the HIF inhibitor acriflavine (ACF) to the mice 3 h before hCG. We found a decrease in ovarian HIF1A and VEGFA, and in the vascular area in the animals treated with ACF. Moreover, we observed an increase in aberrant structures in the ovaries and in luteal cell apoptosis. Serum progesterone levels were decreased together with ovarian STAR expression. However, the animals treated with ACF during the early formation of the corpus luteum were completely fertile and no alterations were observed when the treated females were mated with fertile males. These results collectively suggest that HIF regulates gonadotropin-induced corpus luteum formation acting on luteal blood vessel formation, luteal cell survival, and progesterone synthesis. However, adequate HIF activity may not be essential to achieve and maintain pregnancy. These findings are significant to better understand the complex mechanisms of corpus luteum formation and identify potential abnormalities to allow better knowledge of ovarian physiology and pathologies in which this factor could be involved.

Age at first gestation in bovine influences female progeny post-weaning growth and reproductive development

This study investigated the impact of the age at which heifers conceive for the first time on the growth and reproductive development of their female offspring. A total of seven heifers pregnant at 15 months of age (15M), nine heifers pregnant at 27 months of age (27M) and seven multiparous pregnant cows (Adult) were used in the present trial. All dams were pregnant by AI from a single sire and managed in a single group during gestation, lactation and rearing stages. After weaning heifer calves were stocked on natural pastures. Progeny of heifers that gestated for the first time at different ages did not present differences in growth; however, progeny of 15 and 27M dams had reduced BW, longissimus muscle area and 12th fat thickness compared to progeny of adult dams (P < 0.05). Diameter of the largest follicle was greater in progeny born to Adult compared to progeny born to 15 and 27M (P = 0.04). Reproductive tract score (RTS) in heifers born to Adult at 399 days of age was greater compared to heifers born to 27M, and this category presented greater development than heifers born to 15M (P = 0.02). At 435 days of age, the heifers born to Adult had a greater percentage defined as pubertal compared to the progeny of 15 and 27M (P = 0.04). At 495 days of age, the weight of uterus, ovaries and corpora lutea were not affected by the age of dams at calving (P > 0.25). These data indicated that the age at which heifers received their first service affects female offspring growth RTS at early ages, but it does not have any impact in organ development after diet with a high energy level.

Prolactin locally mediates follicular atresia in hyperprolactinemic vizcachas (Rodentia, Chinchillidae)

Infertility in hyperprolactinemic females is attributed to the dysregulation of GnRH release, subsequently affecting gonadotropin levels, and ultimately leading to anovulation. However, in addition to the hypothalamus, prolactin receptor (PRLR) is expressed in ovaries as well, suggesting potential local effects of PRL in cases of hyperprolactinemia. We have developed an experimental model of sulpiride (SPD)-induced hyperprolactinemia using a wild rodent, the plains vizcacha, and studied the implications of pharmacological PRL levels on folliculogenesis and steroid production. Ovaries of SPD females showed a striking number of atretic follicles along with a reduction in the collective number of viable follicles leading to a higher atretic/viable follicle ratio compared to that of control females (CTL) (P < 0.05). In terms of sensitivity to the hormonal environment, SPD ovaries substantially changed their potential responsiveness to pituitary PRL, as evidenced by the three-fold increase in PRLR expression alongside heightened expression of both gonadotropic receptors in comparison to CTL ovaries (P < 0.05). Circulating estradiol (E2) values doubled post-treatment in the SPD females, which also showed higher expressions of aromatase and 17β-hydroxysteroid dehydrogenase, along with both E2 receptors, ERα and ERβ than the CTL group (P < 0.05). Our findings strongly suggest that hyperprolactinemia-dependent dysregulation of ovarian function can be explained at least partially, by PRL direct actions facilitated by the heightened expression of PRLR in follicles and corpora lutea. Possibly these PRL actions synergize with those triggered by gonadotropic hormones ultimately leading to alteration of the steroidogenic pathway, folliculogenesis disruption and increased atresia.

Ovarian expression of MerTK and its ligand Pros1 in non-pregnant estrus and pregnant mice.

The interaction of MerTK, which negatively regulates immune responses, with its ligand Pros1 contributes to the resolution of apoptosis and inflammation, participating in the healing process of tissues. The levels of MerTK and Pros1, intensely expressed in macrophages (Mϕs), are affected by sex hormones. The expression levels of these proteins in Mϕs, which have a role in corpus luteum (CL) development or regression and folliculogenesis, were investigated in this study since their expressions have not been evaluated in pregnant mouse ovaries. We analyzed mouse ovaries from non-pregnant mice at estrus and gestation days 5, 8, and 15 (each n:10). We used qPCR to evaluate Mertk and Pros1 mRNA levels and assessed their protein expression and localization using immunohistochemistry and double immunofluorescence staining for co-localization. Mertk and Pros1 mRNA and protein levels significantly increased in GD15. MerTK and Pros1 protein levels in mouse CL on GD15 were significantly higher than all other groups. MerTK and Pros1 positive Mϕs were observed in CL of GD15 by double immunofluorescence. MerTK protein levels were increased in granulosa cells GD15 of primary and growing follicles. Our study revealed for the first time that the expression of MerTK and Pros1 was significantly increased in CL at GD15 in mice. These results suggest that increased levels of MerTK andPros1 may enhance their interaction as receptor-ligand binding partners in CL potentially contributing to the balance of apoptosis and inflammation.

Obesity induced by a high-fat diet changes p62 protein levels in mouse reproductive organs.

Obesity is one of the major risk factor for infertility since it causes decreased quality and quantity of gametes and a disrupted uterine environment which might result in miscarriage, stillbirth, and fetal abnormal growth. Obesity induces oxidative stress which is strongly associated with infertility. The clearing of oxidative stress by autophagy is maintained through the p62/ Keap1/Nrf2 pathway. In this pathway, oxidative stress induces p62 for binding to Keap1, thereby Keap1 cannot bind to the Nrf2 transcription factor. Then, Nrf2 translocates into the nucleus and initiates antioxidant-related gene expression. While p62, bound to Keap1, acts as an adaptor protein between autophagosome and damaged substrates which needs to be degraded for homeostasis. Up to date, obesity is strongly linked to abnormal autophagy activity. However, p62 protein expression has not been investigated in the obese ovary, testis, and uterus in detail. Thus, in the present study, we aimed to evaluate the effects of a high-fat diet (HFD)-induced obesity on p62 protein levels of the ovary, testis, and uterus in mice. Our results demonstrated that the p62 expression level was significantly altered by HFD in uterine glands, epithelium, myometrium, and stroma, and in the ovarian corpus luteum, testicular spermatogonium and spermatocytes.

Dietary supplementation with thyme oil improves the reproductive characteristics of Barki adult and prepubertal ewes

The reproductive technology has a significant impact on the development of livestock production. The thyme oil, rich in phytoestrogen chemicals like apigenin, has been found to enhance reproductive performance by mimicking estrogen’s action. This study aimed to investigate the effects of oral supplementation with thyme essential oil on the reproductive organ biometry, and reproductive performance in adult and prepubertal Barki ewes. Seventy ewes were treated with intravaginal sponges impregnated with 40 mg. medroxyprogesterone acetate for 14 days and simultaneously assigned randomly to two groups (20 adult and 15 prepubertal ewes per group), i.e., a control and a thyme oil treated (2.25 mg/kg body weight) group. The number of follicles, follicle diameter, corpora lutea diameter, and estrogen concentration were higher in the thyme oil-treated group than in the control group without comparing between adult ewes and prepubertal ewes. Additionally, the number and diameter of the large follicles were higher (P < 0.05) in the right-side ovary of adult ewes compared with that in prepubertal ewes at day 15. Moreover, thyme oil treatment resulted in higher conception (P < 0.01), lambing rates (P < 0.05), and fecundity (P < 0.01), with values 95.12, 136.73, and 130.25 compared with those in the control group, with values 63.51, 105.24, and 66.97, respectively, without comparing between adult ewes and prepubertal ewes. The adult ewes had a stronger estrus response and better fertility measurement values than prepubertal ewes. Additionally, the estrogen serum levels were positively correlated with the number (P < 0.05) and diameter (P < 0.01) of follicles, and the progesterone serum concentration was positively correlated with the corpora lutea diameter (P < 0.01), without treated group. In conclusion, dietary supplementation with thyme oil improved the follicular population and reproductive performance, which has a good effect on the adult and prepubertal ewes in the Barki ewes.

Shanyao regulates the PI3K/AKT/P21 pathway to promote oogonial stem cell proliferation and stemness restoration to alleviate premature ovarian insufficiency

Ethnopharmacological relevanceShanyao (SY, yam, Rhizoma Dioscoreae, the dried rhizome of Dioscorea oppositifolia L.) was recorded in the Chinese pharmacopoeia and was often used in the treatment of premature ovarian insufficiency (POI). Aim of studyTo evaluate the efficacy of shanyao in cyclophosphamide (CTX)-induced POI and explore its potential mechanism of action. Material and methodsWe employed network pharmacology, Liquid Chromatograph Mass Spectrometer (LC-MS), and molecular docking methods to identify active compounds and core targets, and predict the mechanism of shanyao for treating POI. The mechanism was subsequently validated through a series of experiments. Female Sprague-Dawley (SD) rats were randomly divided into five groups: control (CON), model, estradiol valerate (EV), low-dose shanyao, and high-dose shanyao. An experimental rat model of POI was established using cyclophosphamide and treated with either shanyao or EV for a duration of two months. We assessed the efficacy of shanyao in vivo through methods such as weighing, Enzyme-linked Immunosorbent Assay (ELISA), and Hematoxylin and Eosin (H&E) staining. Oogonial stem cells (OSCs) were isolated, after modeling, treated them with a serum containing either shanyao or EV. Using methods such as CCK8 assay, immunofluorescence staining, flow cytometry (FCM) analysis, and Western blot analysis to verify the mechanism of shanyao in treating POI. ResultsIn this study, we found that after treatment with shanyao, the general condition of POI rats was improved, body weight and the ratio of ovarian weight to body weight were increased, FSH, E2 and AMH levels were improved, primary follicles and preantral follicles were significantly increased, atretic follicles were decreased. However, the number of antral follicles and fresh corpus luteum was no statistical difference. We identified 10 active compounds of shanyao that act on 220 target genes, 176 of which are associated with POI. Denudatin B and Kadsurenone were finally identified as core components. Through topological analysis, 18 key targets were selected, and ultimately PI3K, CCND1, and CDK4 were identified as core targets. Molecular docking results showed that core components had good binding energy with core targets. The results of GO and KEGG enrichment analysis mainly focus on cell cycle regulation and PI3K/AKT signaling pathway. A series of molecular biology experiments confirmed that after shanyao treatment, the phosphorylation level of PI3K and AKT in POI rats were increased, P21 was inhibited, PI3K/AKT/P21 signaling pathway was activated, and the expression levels of CCND1 and CDK4 were increased. At the same time, the expression of Oct4, fragilis and Mvh of ovarian stem cells was up-regulated. ConclusionThe active compounds of shanyao can regulate the PI3K/AKT/P21 signaling pathway, promote the proliferation of oogonial stem cells, stemness restoration, and delay ovarian aging. This study provides valuable insights into shanyao treatment for POI.

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.

Clinical and anamnestic characteristics of patients with functional ovarian cysts (Retrospective study)

Ovarian cysts are common in young women of reproductive age. It is estimated that around 7% of women worldwide experience a symptomatic cyst during their lifetime. Functional ovarian cysts (FOCs) occupy one of the leading places in the structure of gynecological diseases (45-55%) in women of reproductive age. Functional cysts are not true neoplasms and are considered as a variant of a normal physiological process.Ovarian retention formations are found in 7.8% of clinically healthy women and in every second (52.3%) patient with lower abdominal pain or menstrual cycle disorders. Studies of FOCs have mainly concerned cysts that arose after ovulation induction, studies of spontaneous follicular cysts are limited, the question of the effect of FOCs on fertility and ovarian reserve is debated.The objective: to analyze the management of patients with FOCs in real clinical practice.Materials and methods. A retrospective case-control cohort study was conducted with the participation of 350 women aged 19 to 39 years who were diagnosed with ovarian neoplasms between 2020 and 2022. Complaints, anamnesis data, applied methods of diagnosis and treatment of neoplasms were analyzed. Results. According to clinical data, the cases were divided into 3 groups. The I group included 122 (34.9%) women in whom, in the absence of symptoms and complaints, ovarian neoplasms were detected for the first time during pelvic ultrasound.The II group consisted of 116 (33.1%) female patients, who for the first time applied for an examination due to complaints of periodic unilateral pain of various nature, a feeling of discomfort in the lower abdomen.The III group included 112 (32.0%) women who were urgently hospitalized with complaints of acute pain and pressure in the lower abdomen, bloating, weakness, dizziness. Among the patients of this group, 53 (47.3%) women had ovarian neoplasms diagnosed earlier (within 2.6±0.7 years).Among the studied cohort, according to ultrasound data, functional (follicular cysts and corpus luteum cysts) ovarian cysts were verified among the detected neoplasms in 253 (72.3%) women.Conclusions. The prevalence of functional ovarian cysts in women that do not have clinical manifestations as ultrasound findings is 78.7%, among them follicular cysts predominate (66.4%). Infertility occurs in 23.8% of asymptomatic women with functional ovarian cysts. In 34.5% of women with functional ovarian cysts who have clinical symptoms, menstrual cycle disorders prevail.

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.

Fertility and early embryonic development in a CD46-edited Gir heifer with reduced susceptibility to BVDV.

Bovine viral diarrhea virus (BVDV) infection during pregnancy is a significant contributor to reproductive failures in cattle. The bovine receptor for BVDV (CD46) was previously edited with a six amino acid substitution (G82QVLAL to A82LPTFS) and shown to have significantly reduced BVDV susceptibility in a Gir heifer calf. Since a role for CD46 has been proposed in mammalian fertilization, our objective was to assess the edited heifer's fertilization rates, early embryonic development, and germline transmission conformation of the edit. Cumulus oocyte complexes were collected from the edited heifer and unedited females, fertilized with semen from an unedited bull and cultured until the blastocyst stage. Ultrasound examinations and serum progesterone concentration were also monitored to confirm estrous cyclicity in the CD46-edited heifer. Estrous cyclicity was normal with visualization of a corpus luteum and elevated progesterone concentrations. Fertilization rates and blastocyst development were not different in oocytes from edited and unedited controls. Genome sequence analysis of blastocysts confirmed germline transmission of either edited allele from the heifer. Subsequently, the CD46-edited heifer was artificially inseminated with semen from an unedited Gir bull and fertility status was confirmed with a diagnosed conception at day 35 of gestation. Thus, a six amino acid substitution in CD46 did not negatively affect fertilization of edited oocytes or early embryonic development when fertilized with semen from an unedited bull. An edited bull is still needed to similarly evaluate reproductive function of sperm cells carrying this CD46 edit.

The Utilization of a Pregnancy-Associated Glycoprotein Profile and PAG/P4 Ratio Biomarker for the Diagnosis of Pseudopregnancy in Dairy Goats.

Pseudopregnancy (PPG) is a common reproductive disorder in dairy goats, characterized by the accumulation of sterile fluid in the uterus and persistence of a corpus luteum, leading to temporary infertility and reduced farm efficiency. This study aimed to determine the pregnancy-associated glycoprotein (PAG) profile in dairy goats and to evaluate the PAG/P4 ratio as a potential novel biomarker for PPG diagnosis. A total of 605 Saanen and crossbreed mated goats were evaluated via transabdominal ultrasonography between 31 and 52 days after the removal of bucks in four intensive dairy farms, presenting an overall PPG incidence of 7.8%. Blood samples from PPG (n = 47), nonpregnant (NPG; negative control; n = 11), and pregnant (PG; positive control; n = 10) does were collected for PAG and P4 analysis. The PAG plasma levels were 0.08 ± 0.02, 0.13 ± 0.04, and 1.45 ± 0.04 OD (optical density) in the PPG, NPG, and PG groups, respectively (p < 0.001). The PAG/P4 ratio was lower in the PPG group (0.01 ± 0.11; p < 0.001) than in the NPG (0.24 ± 0.23) and PG (0.18 ± 0.23) groups. This difference allows for the establishment of a new methodological diagnostic approach to PPG. The area under the curve (AUC) was 0.996 for a cut-off of 0.04 and 97.9% and 100% sensitivity and specificity, respectively. This study suggests that the PAG/P4 ratio can serve as a reliable biomarker for PPG diagnosis, helping to distinguish it from pregnancy and nonpregnancy and improving reproductive management in dairy goats.

Fertility protective effects of Brillantaisia patula leaf extract against cyclophosphamide-induced ovarian damage in Wistar rats

BackgroundThe primary indication of infertility is the incapacity to conceive, and in females, the majority of instances of female infertility stem from ovulation disorders. This study evaluated the female fertility-enhancing effects and safety of aqueous leaf extract of Brillantaisia patula (ALEBP) in a cyclophosphamide (CYP) model of sterility in Wistar rats.MethodSixty-six female rats randomly allotted to six groups (n = 11) were administered with the appropriate regimen for 21 days and then mated with male rats. Group 1 (control) received distilled water. Groups 2–6 were treated with a single dose (200 mgkg− 1 body weight) of cyclophosphamide intraperitoneally and, in addition, received the same volume (0.5 mL) of distilled water, 18, 36, 72 mgkg− 1 body weight of ALEBP and 200 mg per body weight of vitamin C orally. Mating lasted 11 days; on day 20, the female Wistar rats were sacrificed. Data were analysed using One-way Analysis of Variance (ANOVA) followed by Dunett’s posthoc analysis, and GraphPad (at p < 0.05).ResultsResults herein showed that ALEBP significantly (p < 0.05) increased the diminution in activities/levels of glutathione peroxidase (GPx), reduced glutathione (GSH), total antioxidant capacity (TAC), cholesterol, alkaline phosphatase (ALP), acid phosphatase (ACP), estrogen (ES), and luteinising hormone (LH) induced by cyclophosphamide. ALEBP further reversed the increased level of malondialdehyde (MDA), tumour necrosis factor-α (TNFα), interleukin 8 (IL-8), and follicle-stimulating hormone (FSH) caused by cyclophosphamide (p < 0.05). In addition, ALEBP, while it significantly increased the cyclophosphamide-induced reduction in the number of implantations in each animal, the total number of viable fetuses, the total number of corpora lutea, and the fertility index, also significantly reduced the number of fetal resorptions in each animal and pre-implantation loss that was increased by cyclophosphamide. Moreover, the cyclophosphamide-induced degenerative and necrotic changes in the ovarian cells and uterus were reversed by ALEBP.ConclusionsConsidered as a whole, the aqueous leaf extract of Brillantaisia patula reversed oxidative stress and inflammatory side effects of cyclophosphamide, preserving ovarian function and fertility in the rats. This may suggest its exploration as a safe agent against toxic side effects of chemotherapy and fertility-related disorders of the uterus and ovary.

Transcriptome-based studies on the candidate genes related to body color yellowing in channel catfish (Ictalurus punctatus)

This experiment was aimed to screen the key factors regulating pigment cells and elucidate the potential mechanism of yellowing body color in the channel catfish. Three-channel catfish with all-black body color (NS group) and three channel catfish with yellow body color (YS group) were taken for transcript sequencing analysis using Illumina sequencing technology. The results showed that a total of 2368 significant differentially expressed genes (DEGs) were screened. Compared with the transcription expression genes in the NS group, there were 1003 up-regulated genes and 1365 down-regulated genes in the YS group. Seven DEGs related to pigment regulation were selected for qRT-PCR analysis, and the results confirmed that the transcriptome analysis was reliable. Furthermore, the DEGs were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and the results showed that most of the DEGs were involved in immune response, endocytosis, cell adhesion molecules, etc. Furthermore, analysis of Melanogenesis and Wnt signaling pathways and other pathways with links to pigment regulation obtained by KEGG enrichment revealed that genes that promote melanogenesis, such as etb, plc, and camk2 were significantly down-regulated. Nevertheless, genes that inhibit melanogenesis (nf-κb, tnf-α, ifn) and genes that promote yellow pigmentation (xdh) were significantly up-regulated. In summary, it is hypothesized that oxidative stress is an important factor contributing to the yellowing of the body color of channel catfish, thus causing the expression of genes related to melanogenesis (etb, plc, camk2), yellow pigmentation (xdh), and immune-antioxidant responses (nf-κb, tnf-α, ifn).

The effect of rapamycin treatment on mouse ovarian follicle development in dehydroepiandrosterone-induced polycystic ovary syndrome mouse model.

Polycystic ovary syndrome (PCOS) is a complex reproductive and endocrine disorder affecting 5-10% of women of reproductive age, but the pathophysiology of PCOS still remains unknown. Here, the aim of our study was to analyze the effects of rapamycin treatment that may regulate impaired hormonal levels and folliculogenesis in dehydroepiandrosterone (DHEA)-treated PCOS mouse. We hypothesized that rapamycin may ameliorate the negative effects of PCOS in DHEA-induced PCOS mouse model. The target of rapamycin (TOR) gene product is a serine/threonine kinase that has been implicated in the control of cell growth, proliferation and autophagy, and rapamycin is a potent inhibitor of mTORC1 pathway. In this study, for the first time, mTORC1 and activation products are presented at protein and mRNA levels after rapamycin treatment in DHEA-induced PCOS mouse ovary. We showed that rapamycin treatment may regulate follicular development, hormonal levels and provide ovulation in DHEA-induced PCOS mouse. Additionally, we assessed decreased primordial follicle reserve, increased number of primary and secondary follicles, corpus luteum structure forms again after 10 days of rapamycin treatment. This study presented here suggests rapamycin treatment regulates hormonal phenotype and folliculogenesis in the ovary and also mTOR signalling pathway in granulosa cells of DHEA-induced PCOS mouse ovary which may have potential to attenuate understanding the mechanism of dominant follicle selection and anovulatory infertility.