Zygote最新文献

In cattle, maternal metabolic health has been suggested to influence oocyte and embryo quality. Here, we examined whether maternal liver abnormalities affected in vitro oocyte maturation by screening meiotic maturation, spindle morphology, actin filaments, and lysosomes. In oocytes from the abnormal liver group, the maturation rate (80.2%) was significantly lower compared to a control group with healthy livers (90.8%; P < 0.05). Mean spindle area in oocytes of the abnormal group (50.4 ± 3.4 μm²) was significantly larger than in the control (40.8 ± 1.6 μm²; P < 0.05). Likewise, mean spindle width in the abnormal group (8.8 ± 0.3 μm) was significantly larger than in the control group (7.8 ± 0.2 μm; P < 0.05). The proportion of cells with correctly aligned chromosomes in the abnormal group (48.0%) was significantly lower than in the control (78.3%; P < 0.05). The number of cortical actin filaments in mature oocytes of the abnormal group (299.3 ± 3.7) was significantly lower than in the control (314.7 ± 3.2; P < 0.05). The number of lysosomes in mature oocytes of the abnormal group (1363.6 ± 39.0) was significantly higher than in the control (1123.4 ± 26.3; P < 0.05). In conclusion, our findings indicate that the quality of in vitro matured oocytes is lower in cattle with liver abnormalities than in healthy cattle.

Recently, the World Health Organization recommendation for abstinence time for semen analysis has been challenged in some studies and many of them have supported the advantages of a second short abstinence ejaculation. More evidence is needed to approve this for clinical use. This study aimed to compare the average routine abstinence time (2-7 days) with the short time (1-2 h) on sperm quality based on functional parameters in a population of oligo-astheno-teratozoospermia (OAT) men. The semen samples were retrieved from 50 men with OAT two times: one standard 2-7 days (long ejaculation) and short duration trimming (1-2 hours later the first ejaculation). All semen parameters as well as sperm DNA integrity were compared between groups. Results showed that mean sperm concentration (10.40 vs. 8.76), total sperm count (28.53 vs. 12.24) and mean semen volume (2.69 vs. 1.40) were higher in the first ejaculation (2-7 days of abstinence), while progressive motility (20.52 vs. 13.32), non-progressive motility (53.46 vs. 48.86), morphology (2.46 vs. 1.46) and viability (83.90 vs. 77.96) were significantly higher in the second ejaculation (P < 0.05). The second sample also showed lower immotile (26.82 vs. 38.02) and DNA fragmentation (19.5 vs. 26.96) (P < 0.05). Taking all data into account, an additional short abstinence period (AP) may be a simple and helpful strategy to obtain better sperm quality in couples with male infertility causes, especially in OAT patients. The recommended current guidelines regarding the AP may need to be revisited in severe male factors.

This study explores the efficacy of a novel microfluidic device in isolating rheotactic sperm and assesses their advantages compared with other motile sperm. Two microfluidic devices were used in this study: the microfluidic device we designed to separate sperm based on rheotaxis and a simple passive microfluidic device. We compared the results with the density gradient centrifugation technique. Sperm attributes including concentration, morphology, viability and motility were assessed using related procedures. Statistical analyses were conducted using one-way analysis of variance. Results showed differences in sperm concentration, motility, morphology and vitality using different sperm separation techniques. The sperms separated using our microfluidic device demonstrated the highest motilities, normal morphology percentages and higher sperm vitality but significantly lower sperm concentrations. These findings suggest the potential of our microfluidic design in enhancing sperm quality. Our findings are in agreement with previous research, emphasizing the capability of microfluidics in enhancing sperm quality. Specifically, our designed microfluidic device exhibited exceptional efficacy in isolating highly motile sperm, a critical factor for successful fertilization.

Introduction: Long non-coding RNAs (lncRNAs) are a subset of RNA molecules that have been shown to be involved in gene regulation. A lot of different pathways are involved during gametogenesis and any disturbance to these pathways may have a derogatory impact on producing a haploid gamete and thus a euploid embryo. Steroidogenesis pathway plays a crucial role in gametogenesis. The purpose of this work was to quantify the levels of lnc-CYP11A1-1 and RP11573D15.8 expression levels in aneuploid and euploid embryos. Materials and methods: A total of 20 surplus human embryos, of which 10 euploid and ten aneuploid embryos, were collected from an IVF centre. The expression levels of two lncRNAs, which have been hypothesized to regulate expression of CYP11A1, were evaluated in these embryos. RNA was extracted and used to synthesize cDNA for the experiments. Real-time polymerase chain reaction was performed to evaluate the expression levels of each lncRNA in aneuploid and euploid embryos, respectively. Results and discussion: This study shows that lnc-CYP11A1-1 was more expressed in aneuploid than in euploid embryos. RP11-573D15.8 is expressed more in aneuploid embryos than in euploid ones. The results for RP11-573D15.8 were statistically significant with a p-value of 0.02 (less than the standard threshold of p 0.05), whereas the results for lnc-CYP11A1-1 were not statistically significant with a p-value of 0.07 (greater than the standard threshold of p 0.05). Thus, the result of this study demonstrates that lncRNAs may have a role in gametogenesis and formation of aneuploid gametes.

Rainbow trout (Oncorhynchus mykiss) is a promising cultivable fish species with significant potential for expansion. As a cold-water fish belonging to the Salmonidae family, it requires an optimal temperature range of 10-15°C for optimal growth. This study explores a method for producing sterile rainbow trout with maximum survival rates by using heat shock treatment to enhance growth characteristics and improve aquaculture practices. A control group and four heat shock treatments were given at 26°C and 28°C for 10 min, applied 15 and 20 min after the mixing of eggs and milt, using a water bath. Among the treated groups, the highest fertilisation, hatching and yolk sac absorption rates were 90.3 ± 0.3%, 81.8 ± 0.8% and 83.9 ± 0.5%, respectively. The highest triploidy rate of 76.6 ± 3.3% was observed with a heat shock at 28°C, 20 min after fertilisation. In contrast, none of the fish from the control group were triploids. The control group demonstrated higher survival rates at fertilisation (93.1 ± 0.4%), hatching (84.2 ± 0.4%) and complete yolk sac absorption (86.2 ± 0.5%) compared to the heat-shocked groups. The diploid and triploid chromosome numbers in rainbow trout were determined to be 2n = 60 and 3n = 91, respectively. This study confirms that heat shock treatment can effectively induce triploidy in rainbow trout, with significant variations in triploidy rates depending on the temperature and timing of the shock. While heat shock can enhance the production of sterile fish, it is essential to balance the treatment parameters to maintain high survival rates. These findings contribute to the optimisation of triploidy induction techniques and support the advancement of aquaculture practices by improving the growth, management and survival rates of rainbow trout which could significantly benefit aquaculture efficiency and sustainability.

Treatment with follicle-stimulating hormone (FSH) and testosterone (T2) and their combination have been observed to be influential on ovarian follicles of 1-day-old mice ovaries cultured for 8 days. Given that extension of the culture period could positively impact the development of follicles in cultured ovaries, the present study was conducted to evaluate the main and interaction effects of FSH by T2 on the development of ovarian follicles in 1-day-old mice ovaries cultured for 12 days. One-day-old mice ovaries were initially cultured with base medium for 4 days; thereafter, different hormonal treatments were added to the culture media, and the culture was continued for 8 additional days until day 12. Ovaries were collected for histological and molecular assessments on day 12. The greatest activation of primordial follicles and progression of activated follicles to the preantral stage was detected in ovaries treated with the combination of FSH and T2 (P < 0.05). This positive effect on the morphology of ovarian follicles was accompanied by upregulation of Pi3k, Gdf9, Bmp15, Cx37 and Fshr in the ovaries cultured with the combination of FSH and T2 (P < 0.05). Nonetheless, treatment with FSH and T2 led to a diminished proportion of intact follicles (P < 0.05), even though Bax/Bcl2 gene expression ratio, as an apoptotic index, was less in hormone-treated ovaries (P < 0.05). In conclusion, the combination of FSH and T2 could improve the activation of primordial follicles and the growth of activated follicles towards the preantral stage. This positive effect of FSH plus T2 appeared to be at least partly mediated through the upregulation of Pi3k and oocyte-derived growth factors including Gdf9 and Bmp15.

Growth differentiation factor 9 (GDF9) is an oocyte-specific paracrine factor involved in bidirectional communication, which plays an important role in oocyte developmental competence. In spite of its vital role in reproduction, there is insufficient information about exact transcriptional control mechanism of GDF9. Hence, present study was undertaken with the aim to study the expression of basic helix-loop-helix (bHLH) transcription factors (TFs) such as the factor in the germline alpha (FIGLA), twist-related protein 1 (TWIST1) and upstream stimulating factor 1 and 2 (USF1 and USF2), and nuclear receptor (NR) superfamily TFs like germ cell nuclear factor (GCNF) and oestrogen receptor 2 (ESR2) under three different in vitro maturation (IVM) groups [follicle-stimulating hormone (FSH), insulin-like growth factor-1 (IGF1) and oestradiol)] along with all supplementation group as positive control, to understand their role in regulation of GDF9 expression. Buffalo cumulus-oocyte complexes were aspirated from abattoir-derived ovaries and matured in different IVM groups. Following maturation, TFs expression was studied at 8 h of maturation in all four different IVM groups and correlated with GDF9 expression. USF1 displayed positive whereas GCNF, TWIST1 and ESR2 revealed negative correlation with GDF9 expression. TWIST1 & ESR2 revealing negative correlation with GDF9 expression were found to be positively correlated amongst themselves also. GCNF & USF1 revealing highly significant correlation with GDF9 expression in an opposite manner were found to be negatively correlated. The present study concludes that the expression of GDF9 in buffalo oocytes remains under control through the involvement of NR and bHLH TFs.

EXOSC10 is an exosome-associated ribonuclease that degrades and processes a wide range of transcripts in the nucleus. The initial segment (IS) of the epididymis is crucial for sperm transport and maturation in mice by affecting the absorption and secretion that is required for male fertility. However, the role of EXOSC10 ribonuclease-mediated RNA metabolism within the IS in the regulation of gene expression and sperm maturation remains unknown. Herein, we established an Exosc10 conditional knockout (Exosc10 cKO) mouse model by crossing Exosc10 ^F/F mice with Lcn9-Cre mice which expressed recombinase in the principal cells of IS as early as post-natal day 17. Morphological and histological analyses revealed that Exosc10 cKO males had normal spermatogenesis and development of IS. Moreover, the sperm concentration, morphology, motility, and frequency of acrosome reactions in the cauda epididymides of Exosc10 cKO mice were comparable with those of control mice. Thus, Exosc10 cKO males had normal fertility. Collectively, our genetic mouse model and findings demonstrate that loss of EXOSC10 in the IS of epididymis is dispensable for sperm maturation and male fertility.

This study aimed to demonstrate the utilization value of 1PN embryos. The 1PN zygotes collected from December 2021 to September 2022 were included in this study. The embryo development, the pronuclear characteristics, and the genetic constitutions were investigated. The overall blastocyst formation and good-quality blastocyst rates in 1PN zygotes were 22.94 and 16.24%, significantly lower than those of 2PN zygotes (63.25 and 50.23%, respectively, P = 0.000). The pronuclear characteristics were found to be correlated with the developmental potential. When comparing 1PN zygotes that developed into blastocysts to those that arrested, the former exhibited a significantly larger area (749.49 ± 142.77 vs. 634.00 ± 119.05, P = 0.000), a longer diameter of pronuclear (29.81 ± 3.08 vs. 27.30 ± 3.00, P = 0.000), and a greater number of nucleolar precursor body (NPB) (11.56 ± 3.84 vs. 7.19 ± 2.73, P = 0.000). Among the tested embryos, the diploidy euploidy rate was significantly higher in blastocysts in comparison with the arrested embryos (66.67 vs. 11.76%, P = 0.000), which was also significantly higher in IVF-1PN blastocysts than in ICSI-1PN blastocysts (75.44 vs. 25.00%, P = 0.001). However, the pronuclear characteristics were not found to be linked to the chromosomal ploidy once they formed blastocysts.In summary, while the developmental potential of 1PN zygotes is reduced, our study shows that, in addition to the reported pronuclear area and diameter, the number of NPB is also associated with their developmental potential. The 1PN blastocysts exhibit a high diploidy euploidy rate, are recommend to be clinically used post genetic testing, especially for patients who do not have other 2PN embryos available.

In vitro production of porcine embryos is a complicated process that includes in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). Insufficient cytoplasmic maturation, slow zona reaction and improper embryo culture conditions will compromise the efficiency of porcine embryo production in vitro. Previous studies have shown that insulin-transferrin-selenium (ITS) in IVM or IVC medium could improve porcine oocyte maturation, decrease polyspermy fertilization and promote subsequent embryonic development in vitro. However, the effect of ITS both in IVM and IVC media on porcine embryo production in vitro hasn't been elucidated. In this study, we found that 1.0% ITS supplementation in IVM/IVC media promoted the expansion of cumulus cells, raised mitochondrial membrane potential, increased ATP content and reduced ROS level in matured oocytes, improved blastocyst rate and the cell number of blastocyst, simultaneously. In conclusion, the IVM/IVC media supplemented with 1.0% ITS can improve the efficiency of porcine embryo production in vitro.