Molecular Reproduction and Development最新文献

In the ovaries, cyclic adenosine 3′,5′-monophosphate (cAMP) is a second messenger supporting the generation of steroids. Phosphodiesterases (PDEs) are regulators of intracellular cAMP, and therefore, potential regulators of ovarian function. Interestingly, the family of PDE genes are differentially expressed in human oocytes and granulosa cells from primordial and primary follicles, suggesting diverse roles. In this study, we addressed the functions of PDE3B and PDE8B in primordial follicle regulation using inhibitors of PDE3B and PDE8B in murine ovary primary in vitro cultures. Inhibition of PDE8B in ovarian cultures prevented primordial follicle activation, while inhibition of PDE3B had no effect on follicle distribution in the ovary, under the tested conditions. As cAMP levels may increase steroid levels, we assessed the protein levels of the steroidogenic acute regulatory protein (StAR) and aromatase enzymes, and found that inhibition of PDE3B reduced StAR protein levels, whereas inhibition of PDE8 did not alter StAR expression in our murine ovary culture system conditions. Our results showed that ketotifen-induced inhibition of PDE8B can decrease primordial follicle activation, whereas we observed no effect of follicle distribution, when PDE3B was inhibited. Expression of the StaR enzyme was not altered when PDE8B was inhibited, which might reflect not sufficient inhibition by ketotifen to induce StAR alterations, or redundant mechanisms.

Throughout the reproductive life of women, cumulus cells (CC) protect the dormant oocyte from damage, act as sensors of the follicular microenvironment, and act as a gatekeeper for oocyte developmental potential. One such mechanism relies on the hypoxia-tolerance response, which, with age, decreases systematically, including in the ovary. We aimed to evaluate the association between gene expression related to hypoxia and aging in CC and reproductive results in in vitro fertilization cycles. We recruited 94 women undergoing controlled ovarian stimulation. Total RNA was extracted from pooled CCs collected after oocyte pick-up (OPU) and reverse-transcribed to complementary DNA using random hexamers to test 14 genes related to hypoxia response via HIF1α activation, oxidative stress, and angiogenic responses. The expression of CLU, NOS2, and TXNIP had a positive correlation with age (r_s = 0.25, r_s = 0.24, and r_s = 0.35, respectively). Additionally, NOS2 and HMOX1 expression correlated positively with the retrieval of immature oocytes (r_s = 0.22 and r_s = 0.40, respectively). Moreover, VEGFC levels decreased overall with increasing fertilization rate, independently of age (r_s = −0.29). We found that the fertilization potential of a cohort of oocytes is related to the ability of CC to respond to oxidative stress and hypoxia with age, pointing at NOS2, HMOX1, and VEGFC expression as markers for oocyte maturation and fertilization success.

Preeclampsia is an obstetric disorder and remains the leading contributor to maternal and fetal morbidity and mortality. This study was designed to explore the role of hsa_circ_0001740 in preeclampsia as well as its underlying mechanism. Real-time quantitative polymerase chain reaction was performed to examine hsa_circ_0001740 and miR-188-3p levels in trophoblast cell line HTR-8/SVneo. The proliferation, migration, invasion, and apoptosis of HTR-8/SVneo cells were detected using cell counting kit-8, colony formation, wound healing, transwell, and terminal-deoxynucleoitidyl transferase mediated nick end labeling assays, respectively. The expression of apoptosis- and Hippo signaling-related proteins were assessed by western blot. Moreover, the binding relationship between hsa_circ_0001740 and miR-188-3p, miR-188-3p and ARRDC3 were verified by luciferase report assay. The results showed that hsa_circ_001740 overexpression inhibited the proliferation, migration, and invasion, and promoted apoptosis of HTR-8/SVneo cells. Hsa_circ_0001740 was verified to bind to miR-188-3p, and ARRDC3 was demonstrated to be a target of miR-188-3p. miR-188-3p overexpression partially counteracted the suppressive effects of hsa_circ_001740 overexpression on the proliferation, migration, and invasion of HTR-8/SVneo cells. What's more, ARRDC3 expression was upregulated by hsa_circ_001740-overexpression but was downregulated by miR-188-3p overexpression. Hsa_circ_001740/miR-188-3p also mediated Hippo signaling. To summarize, hsa_circ_0001740 could maintain trophoblast cell function via downregulating miR-188-3p, providing a potential biomarker for the diagnosis and treatment of preeclampsia.

Meiotic defects in oocytes are the primary reason for decreased female fertility with advanced maternal age. In this study, we revealed that decreased expression of ATP-dependent Lon peptidase 1 (LONP1) in aged oocytes and oocyte-specific depletion of LONP1 disrupt oocyte meiotic progression accompanying with mitochondrial dysfunction. In addition, LONP1 downregulation increased oocyte DNA damage. Moreover, we demonstrated that splicing factor proline and glutamine rich directly interacts with LONP1 and mediate the effect of LONP1 depletion on meiotic progression in oocytes. In summary, our data suggest that decreased expression of LONP1 is involved in advanced maternal age-related meiosis defects and that LONP1 represents a new therapeutic target to improve aged oocyte quality.

In many multicellular organisms, mature gametes originate from primordial germ cells (PGCs). Improvements in the culture of PGCs are important not only for developmental biology research, but also for preserving endangered species, and for genome editing and transgenic animal technologies. SMAD2/3 appear to be powerful regulators of gene expression; however, their potential positive impact on the regulation of PGC proliferation has not been taken into consideration. Here, the effect of TGF-β signaling as the upstream activator of SMAD2/3 transcription factors was evaluated on chicken PGCs' proliferation. For this, chicken PGCs at stages 26−28 Hamburger−Hamilton were obtained from the embryonic gonadal regions and cultured on different feeders or feeder-free substrates. The results showed that TGF-β signaling agonists (IDE1 and Activin-A) improved PGC proliferation to some extent while treatment with SB431542, the antagonist of TGF-β, disrupted PGCs' proliferation. However, the transfection of PGCs with constitutively active SMAD2/3 (SMAD2/3CA) resulted in improved PGC proliferation for more than 5 weeks. The results confirmed the interactions between overexpressed SMAD2/3CA and pluripotency-associated genes NANOG, OCT4, and SOX2. According to the results, the application of SMAD2/3CA could represent a step toward achieving an efficient expansion of avian PGCs.

The goal for the present study was to investigate the effect of aneuploidy on embryo morphokinetics events in a time-lapse imaging (TLI) system incubator. This retrospective cohort study was performed in a private university-affiliated in vitro fertilization center, between 2019 March and December 2020. Kinetic data were analyzed in 935 embryos, derived from 316 patients undergoing intracytoplasmic sperm injection cycle with preimplantation genetic testing (PGT) for aneuploidy, individually cultured in a TLI incubator until Day 5 of development. Timing of morphokinetic variables, the incidences of multinucleation, and Known Implantation Data Score (KIDScore)-Day 5 were compared between euploid (n = 352) and aneuploid embryos (n = 583). Aneuploid embryos showed significantly longer timing to complete specific morphokinetic parameters compared to euploidy embryos. Euploidy embryos also showed a significantly higher KIDScore when compared with the aneuploidy ones. Our evidence suggests that TLI monitoring may be an adjunct approach to select embryos for PGT; however, cautious investigation is still needed.

Prediction of a boar's fertility level has great economic importance for sow herds. After standard sperm morphology and motility metrics are met, approximately 25% of boars have less than 80% conception rates. Due in part to the many factors involved in the fertilization process, a multifactorial model incorporating multiple relevant sperm physiology factors will likely lead to increased understanding of boar fertility. Here we review the current literature on boar sperm capacitation as a predictor of boar fertility. While limited, several studies have provided correlations between the percentage of sperm in an ejaculate that are capable of undergoing sperm capacitation in a chemically defined media and artificial insemination field fertility as well as proteome and other methods. Work summarized here underscores the need for further understanding of boar fertility.

Herein we aimed at exploring mitochondrial energy metabolism status in patients with repeated implantation failure (RIF) and whether key regulatory factor PGC-1α of energy metabolism is involved in the decidualization of endometrial stromal cells. Mitochondrial oxidative phosphorylation level and ATP synthesis were compared in primary endometrial stromal cells from RIF and control group. At the same time, as one of key transcription regulators of mitochondrial energy metabolism, the expression level and acetylation level of PGC-1α were compared with two groups. Then, we downregulated the acetylation levels of PGC-1α, and the expression of decidual markers (PRL and IGFBP1) was observed further. Mitochondrial energy metabolism, showing by mitochondrial oxidative phosphorylation level and ATP synthesis, was decreased in the endometrial stromal cells of the RIF group (RIF-hEnSCs). Meanwhile, PGC-1α acetylation levels were significantly higher in RIF-hEnSCs. When we reduced the acetylation levels of PGC-1α in RIF-hEnSCs, the basal O₂ consumption rate and maximal respiration were increased, and also the PRL and IGFBP1. Overall, our data indicated that the endometrial stromal cells of the RIF patients had low level of mitochondrial energy metabolism. Reducing acetylation level of key energy metabolism regulator PGC-1α can increase the decidualization level of RIF-hEnSCs. These findings may inspire new ideas about the treatment of RIF.

Hyper-prolific sows frequently do not have a sufficient number of functional teats for their piglets to nurse which has led to the use of nurse sows to manage these surplus piglets. This review discusses strategies for using nurse sows and factors that influence preweaning survival and weight gain of their litters, as well as those that affect their subsequent rebreeding performance. Rearing piglets using a nurse sow can be as successful as piglets reared with their biological mother and is thus a powerful management tool to decrease preweaning piglet mortality. Selecting a young sow as nurse sow is beneficial for piglet survival; however, piglets nursing first parity sows often have a lower daily weight gain than piglets nursing multiparous sows. A litter of uniform surplus piglets is preferably handled using the two-step nurse sow strategy. A consequence of nonuniform litters will most likely be an increased mortality and decreased weaning weight among the smallest piglets within a litter. The subsequent fertility of nurse sows is not compromised. There is an increased risk of lactational oestrus when using nurse sows leading to an increased weaning-to-oestrus interval; however, litter size in nurse sows is identical or even moderately higher in the subsequent parity compared with nonnurse sows.

This work aims to study the testicular morphology and spermatogenesis of Gymnotus carapo to provide information on their reproductive biology which is useful in managing this species as a fishing resource. The testicles were isolated and fixed in 10% formalin; subsequently, they were processed for scanning electron microscopy with conventional histological technique. To analyze the cell proliferation of germline cells and Sertoli cells, immunodetection of the proliferating cell nuclear antigen (PCNA) protein was performed. In G. carapo spermatogenesis, the spermatogenic line is organized into cysts. Spermatogonia A is characterized by more bigger and solitary cells. Spermatogonia B are smaller cells; their nucleus has a larger area concerning the cytoplasm and is grouped in tubules. Spermatocytes (I–II) are smaller than spermatogonia in the prophase of meiotic division. Spermatids are cells with dense, rounded nucleus. The sperm were found in the lumen of the tubule. By immunostaining PCNA, it was possible to observe the proliferative activity of germ line cells and Sertoli cells during the cyst reorganization phase. These results are the basis for future studies focusing on the analysis compared to females of the reproductive cycle of G. carapo.