Biology of Reproduction最新文献

Altered peristaltic and ciliary dysfunction is a feature of females with endometriosis. To further explore this premise, we examined the ampulla of rhesus macaques (Macaca mulatta) with and without spontaneous endometriosis for the expression of adenylate kinase 7 (AK7), a mitochondrial-dwelling nucleotide converting enzyme with critical roles in cellular kinesis, forkhead protein box J1 (FOXJ1), a marker of cilia abundance, and Anoctamin 1 (ANO1) as a marker of both smooth muscle contraction and ciliogenesis. We further performed an in vitro experiment that treated ampullary segments with peritoneal fluid from animals with and without endometriosis. We report significantly downregulated expression of ANO1 in the ampulla of monkeys with endometriosis (in vivo), and in the ampullary segments exposed to peritoneal fluid of animals with endometriosis. We did not observe statistically significant differences in the expression of AK7 or FOXJ1 both in vivo and in vitro. This highlights potentially essential roles of ANO1 in the oviduct, the dampening of which may lead to a specific subtype of endometriosis-caused subfertility.

Endometriosis is an estrogen dependent disease, which is related to infertility. Decidualization is a prerequisite for successful implantation of human embryos, and endometriosis affects the occurrence of decidualization. However, the mechanism that affects decidualization in endometriosis is not fully understood. Here, we find that Aurora kinase A (AURKA) is upregulated in the eutopic endometrium of endometriosis. AURKA inhibits the decidualization of stromal cells in the eutopic endometrium of endometriosis. Furthermore, in animal experiments, AURKA promotes endometriosis and inhibits decidualization in mice with endometriosis, leading to decreased expression of decidualization markers, such as prolactin, insulin-like growth factor-binding protein-1, and desmin. Afterwards, we find that nuclear factor-κB (NF-κB) p65 is a new substrate of AURKA. AURKA interacts with p65 to promote its phosphorylation and nuclear translocation. Meanwhile, AURKA enhances the protein stability of p65 by prolonging its half-life. In summary, AURKA inhibits the decidualization of the eutopic endometrium in patients with endometriosis by regulating p65, which may provide new ideas for improving decidualization defect in patients with endometriosis.

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.

Natural ovarian aging is one of the major causes for declining fertility in female animals, which has become an insurmountable issue in human reproduction clinics and assisted reproductive technology procedures. Nevertheless, the molecular basis of oocyte aging remains poorly understood, and feasible improvement strategies are unavailable. In the present study, in vivo supplementation of pyrroloquinoline-quinone effectively elevated the fecundity of reproductively aged mice by balancing hormonal secretion, harmonizing the estrus cycle, and eliminating ovarian fibrosis. Moreover, oocyte quality also increased in aged mice after pyrroloquinoline-quinone administration from various aspects, including nuclear and cytoplasmic maturation competency, fertilization capacity, and pre-implantation embryonic development potential. Transcriptomic analysis identified target pathways that might mediate pyrroloquinoline-quinone's effects in aged oocytes. Specifically, it was demonstrated that pyrroloquinoline-quinone supplementation restored the mitochondrial dynamics and lysosomal function to remove excessive reactive oxygen species and suppress apoptosis in aged oocytes. Jointly, these findings demonstrate pyrroloquinoline-quinone administration is an efficacious method to restore the compromised ovary function and damaged oocyte quality in reproductively aged mice, which might be a potential clinical therapy for women of advanced maternal age with infertility.

Reactive oxygen and nitrogen species (RONS) are essential components of diverse intracellular signalling pathways. In addition to their involvement in apoptosis, RONS are crucial in the regulation of multiple developmental and physiological processes. This review aims to summarise their role in the regulation of key ovarian stages: ovulation, maturation and postovulatory ageing of the oocyte, and the formation and regression of the corpus luteum. At the cellular level, a mild increase in RONS is associated with the initiation of a number of regulatory mechanisms, which might be suppressed by increased activity of the antioxidant system. Moreover, a mild increase in RONS has been linked to the control of mitochondrial biogenesis and abundance in response to increased cellular energy demands. Thus, RONS should also be perceived in terms of their positive role in cellular signalling. On the other hand, an uncontrolled increase in ROS production or strong-down regulation of the antioxidant system results in oxidative stress and damage of cellular components associated with ovarian pathologies and ageing. Similarly, the disturbance of signalling functions of RNS caused by dysregulation of NO production by NO synthases in ovarian tissues interferes with the proper regulation of physiological processes in the ovary.

Germ cell mutagenicity testing is increasingly required for chemical risk assessment. Duplex Sequencing (DS) is rapidly gaining acceptance as a method to assess in vivo mutagenesis, and as a valid alternative to transgenic rodent (TGR) mutation models such as the MutaMouse. We used a DS panel of 20 genomic targets and the TGR assay to measure mutations in the germ cells of MutaMouse males exposed to 0, 1, 2, or 5 mg/kg N-ethyl-N-nitrosourea (ENU) for 28 days. Germ cells from the seminiferous tubules were collected 28 days post-exposure. The TGR assay showed a significant increase in mutant frequencies at the high (p < 0.001) and medium (p = 0.01) ENU doses relative to controls, while DS revealed a significant increase (p < 0.001) in ENU-induced mutations only at the high-dose. DS mutation frequencies were lower in genic than in intergenic targets, suggesting a protective role for transcription-coupled repair. Interestingly, we observed several unique germ cell characteristics with respect to DS data from rodent somatic tissues: 1) larger inter-animal variability in clonally expanded mutations that affects the ability to detect significant increases in mutation frequency; 2) a target on chromosome 2 showing much higher susceptibility to spontaneous and chemical-induced mutagenesis than other targets; and 3) a mutation spectrum consistent with that observed in the offspring of ENU-treated males but not with the spectrum in bone marrow of directly-exposed males. These results suggest that DS is a promising approach for characterizing germ cell mutagenesis and that mutagenic mechanisms operating in germ cells differ from those in somatic tissues.

Kynurenine (KYN) is a primary tryptophan derivative found in the human body and fermented foods. Previous studies have shown that KYN is an Aryl hydrocarbon receptor (AHR) agonist and is important in regulating various physiological activities, including female reproduction. Progesterone is a vital steroid hormone that facilitates embryo implantation and maintains pregnancy. However, whether KYN affects its biosynthesis remains unclear. To gain understanding, in vitro luteinized porcine granulosa luteal (pGL) cells were treated with KYN. The results showed that KYN disrupted progesterone biosynthesis by decreasing the expression of STAR and HSD3B in pGL cells. In addition, the expression of three transcription factors of STAR and HSD3B (GATA4, GATA6, and CEBPB) decreased after KYN treatment. Furthermore, the AHR blockade results showed comparable to those of KYN treatment, and subsequent knockdown experiments confirmed these results. These findings suggest that KYN inhibits progesterone biosynthesis in pGL cells by downregulating GATA4, GATA6, and CEBPB expression through AHR. Thus, our results showed for the first time, a previously unknown connection between KYN and progesterone biosynthesis.

The purinergic system is composed of purine nucleotides, enzymes, transporters, and receptors and involved in a variety of physiological processes in the body, including development, metabolism, immunity, tumorigenesis, and reproduction. The importance of the purinergic system for embryo implantation in the endometrium and in pathophysiological conditions has been shown in some species. However, the expression, regulation, and function of purinergic system molecules at the maternal-conceptus interface in pigs is not fully understood. Therefore, we determined the expression of purinergic system molecules in the endometrium during the estrous cycle and pregnancy and in the conceptus and chorioallantoic tissues during pregnancy in pigs. The expression of many purinergic system molecules in the endometrium changed dynamically during pregnancy, with the expression of several components in the endometrium greater on Day 15 of pregnancy than Day 15 of the estrous cycle, and it was regulated by conceptus-derived interleukin-1β and interferon-γ. Purinergic system molecules were also expressed in conceptuses during early pregnancy and in chorioallantoic tissues during mid- to term pregnancy. Furthermore, ATP, a major purinergic signaling molecule, increased migration of endometrial epithelial and conceptus trophectoderm cells, the expression of prostaglandin synthetic enzymes, and the secretion of prostaglandin F2α in endometrial epithelial cells in vitro. These data suggest that the purinergic system molecules expressed in the endometrium, conceptus, and chorioallantoic tissues might play an important role in the establishment and maintenance of pregnancy by regulating various cellular functions at the maternal-conceptus interface in pigs.

Diet-induced obesity can alter ovarian functions after long-term high-fat high-sugar (HFHS) diet feeding. However, the acute impact of obesogenic diets on the follicular cells, and how this impact evolves over time when intake is continued and obesity develops, is not known. Our aim was to determine the onset and progression of changes in the granulosa cell (GC) transcriptomic profile after starting a HFHS-diet feeding in mice. We also examined the changes in oocyte lipid droplet content (LDC) and mitochondrial ultrastructural abnormalities. Swiss (outbred) mice were sacrificed at 24 h, 3 days, and at 1, 4, 8, 12 and 16 wks of feeding HFHS and control diets. LDC significantly increased in the HFHS oocytes already after 24 h compared to controls (P < 0.05). Oocyte mitochondrial abnormalities only increased starting from 8wks. Granulosa RNA-seq revealed altered transcriptomic gene-set enrichments (GO-terms and KEGG pathways, P adj < 0.05) already at 3d and 1wk indicating acute endoplasmic-reticulum unfolded-protein responses, with concomitant fluctuations in several cellular metabolic pathways and gene-sets related to mitochondrial bioenergetic functions, some of which persisted after 8wks. Interestingly the short- and long-term patterns of changes in cytochrome P450, steroid hormone biosynthesis, retinol metabolism, bile acid metabolism, fatty acid metabolism and Pi3K/Akt signaling pathways were most prominent and highly correlated; all being acutely upregulated, then chronically downregulated. These results show that the impact of obesogenic diet on the oocyte and granulosa cells is prompt, while the response depends on the duration of feeding and occurs in a multiphasic cascade together with a progressive deterioration in oocyte quality.

Infertility and post-partum reproductive diseases are significant challenges in cattle farming, with the maternal immune system's ability to recognize and tolerate the embryo being crucial for successful gestation. DNA methylation in hematopoietic cells may influence susceptibility to post-partum fertility issues, making the identification of epigenetic changes vital for sustainable animal production. This study aimed to characterize the methylome of immune cells in relation to fertility, potentially enabling early detection of subfertility. Using whole epigenome sequencing and enzymatic methyl-seq, we analyzed DNA methylation patterns in blood from twelve Holstein cows before the onset of any disease. Our findings revealed 216,990 differentially methylated cytosines (DMCs) between fertile and subfertile cows. Notably, three genes-Interferon tau-3 (IFNT3), KIAA0825, and RAS-Related Protein 2A-showed high significance in their differential methylation between fertile and subfertile cows. IFNT3, crucial for early embryonic development, had seven DMCs in its TSS shores in subfertile cows. Additionally, the KLRA1 gene (Ly49), was identified as containing DMCs across all five genomic regions analyzed (TSS shores, exons, introns, downstream, and distal intergenic). Its widespread differential methylation highlights its potential impact on fertility. Key interleukin genes, including IL6, IL15, IL22, and IL36G, also showed multiple DMCs, reinforcing the role of the immune system in bovine fertility. These findings illustrate the potential control that immune cell epigenetics exert on cattle post-partum fertility. Additionally, this study suggests that the risk of developing subfertility could potentially be estimated with as few as 220 biomarkers, paving the way for enhanced animal health management and improved fertility treatments.