Endometrial receptivity is the ability of the endometrium to accept embryos. Thus, endometrial receptivity dysfunction is an important factor leading to embryo implantation failure. A good endometrial receptivity provides a suitable environment for embryo implantation, improving the embryo implantation rate. The “implantation window” stage, or the receptive stage of the endometrium, is regulated by various hormones, genes, proteins and cytokines, among which microRNAs (miRNAs) and their target genes have a regulatory effect on endometrial receptivity. This review outlines the relationship between endometrial receptivity and pregnancy, the mRNAs and related signalling pathways that regulate endometrial receptivity, and the regulatory role of miRNA in endometrial receptivity, providing a deeper understanding of the regulatory mechanisms of miRNA on endometrial receptivity in humans and animals and reference for the endometrial receptivity-related research.

One of the most recognizable cases of preimplantation genetic diagnosis (PGD) is X-linked diseases. Diagnosis of fetal sex is essential for couples who are known to be at risk of some X-linked disorders. The objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting sex chromosomes-specific sequences in spent culture medium and comparing these results to PGD/CGH array results. It may open new window for the development of a non-invasive PGD method. 120 Embryo’s spent media from Day 3 and Day 5 embryos were collected. Modified phenol-chloroform solution was used for DNA extraction from spent media. Sex determination was performed using SRY, TSPY and AMELOGENIN evaluation through quantitative polymerase chain reaction (q-PCR) method. IBM SPSS and MedCalc were used for statistical analyses to compare sex determination of embryos by spent medium with PGD/CGH array results. Culture time was demonstrated to increase the DNA amount among day 5 embryos culture medium samples. Non-invasive PGD by means of spent culture medium gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% for sex determination. Results of sex determination using spent medium by q-PCR were consistent with the results of PGD/CGH array. Improvements in cell-free DNA extraction and PCR amplification procedures provide us an effective method to perform a PGD test without biopsy in the future, especially about X-linked diseases.

This study aimed to analyze the influence of different temperatures on the embryonic and larval development of discus fish Symphysodon aequifasciatus and determine the time required for the beginning of exogenous feeding. Eggs and larvae were obtained from natural spawns and distributed in five treatments: 24.0, 26.0, 28.0, 30.0, and 32.0 °C. To assess the developmental stages and embryonic structures, samples were taken at regular intervals and checked under an optical microscope. At the end of the experimental period, statistical analysis was performed, followed by Tukey’s test. As a result, it was possible to observe the significant effects of temperature on the variables. It was noted that the temperature accelerated the embryonic and larval development of the discus and also contributed to a reduction in the time between the incubation period and the feeding transition. It was also noted that the incubation of eggs and larvae at a temperature of 24.0 °C can cause damage to embryos, such as malformation of the body as well as anomalies in the circulatory system.

We herein investigated the influence of temperature on the embryonic development (from fertilisation to hatching) of Mugil liza larvae. For this purpose, oocytes (>600 μm) and sperm were obtained from breeding stock at the laboratory of marine fish culture (LAPMAR). After fertilisation, 1200 eggs were distributed in 12 cylindrical experimental units of 400 mL under four different temperatures 18, 22, 26 and 30 ºC, all in triplicate. Every 15 min until hatching, about 10 eggs were randomly sampled in each treatment. The eggs were visualized and photographed, and the classification of embryonic stages was performed. Temperature influenced the main events of the embryonic development of M. liza. More accelerated development was observed according to the increase in temperature until the gastrula phase. At temperatures of 22 and 26 °C, embryonic development occurred from fertilisation to hatching of the larvae. In the 18 °C treatment, it was verified that most of the embryos ceased development during the final phase of cleavage and the beginning of blastula formation, while in the 30 °C treatment patterns of embryo malformation were also verified, with erratic divisions of the blastomeres, resulting in irregular cells. Unlike what was observed at a temperature of 18 °C, none of the embryos incubated at 30 °C reached the blastopore closure phase, stopping in the gastrula. The larvae hatched in the treatments at 22 and 26 °C were viable and exhibited intense swimming, with a large amount of reserve material (yolk) and an evident drop of oil.

The aims of this study were to evaluate the doxorubicin concentration that induces toxic effects on in vitro culture of isolated mouse secondary follicles and to investigate whether resveratrol can inhibit or reduce this toxicity. Secondary follicles were isolated and cultured for 12 days in control medium (α-MEM+) or in α-MEM+ supplemented with doxorubicin (0.1 µg/ml) or different concentrations of resveratrol (0.5, 2, or 5 µM) associated with doxorubicin (0.1 µg/ml) (experiment 1). For experiment 2, follicles were cultured in α-MEM+ alone or supplemented with doxorubicin (0.3 µg/ml) or different concentrations of resveratrol (5 or 10 µM) associated or not with doxorubicin (0.3 µg/ml) (experiment 2). The endpoints analyzed were morphology (survival), antrum formation, follicular diameter, mitochondrial activity, glutathione (GSH) levels and DNA fragmentation. In the first experiment, doxorubicin (0.1 µg/ml) maintained survival and antrum formation similar to the control, while 5 µM resveratrol showed increased parameters, maintained mitochondrial activity and increased GSH levels compared to the control. In the second experiment, doxorubicin (0.3 µg/ml) reduced survival, antrum formation and follicular diameter compared to the control. Resveratrol at a concentration of 10 µM attenuated the damage caused by doxorubicin by improving follicular survival and did not present DNA fragmentation. In conclusion, supplementation of the in vitro culture medium with 0.3 µg/ml doxorubicin reduced the survival and impaired the development of mouse-isolated preantral follicles. Resveratrol at 10 µM reduced doxorubicin-induced follicular atresia, without DNA fragmentation in the follicles.

Although many Fertility Centers have adopted day 5 or 6 embryo transfer policy, yet, 30% of embryo transfers in the US are performed on day 3. This is mainly due to concerns related to longer embryo culture effect and higher rates of embryo transfer cancellation on day 5, with no effect on cumulative pregnancy rate. We conducted a retrospective cohort study comparing individual embryo transfer order rank, best embryo for fresh transfer and intention to freeze, of day-3 and day-5 embryos based on their morphology score. Day-3 embryos of each patient were ranked by embryologists for the order of transfer and intention to freeze, based on morphological score, blinded to actual blastulation outcome. The corresponding blastocysts were similarly ranked for the order of transfer and vitrification intention. Ranking was compared to test the predictive value of day-3 morphological assessment. Sixty patients with 784 day-3 embryos were included. There was only a moderate positive significant correlation between ranks on day-3 and ranks on day-5 [r = 0.662 95% CI (0.611–0.706, p < 0.001)]. Only 25% of the best embryos for transfer on day 3 (rank = 1) were chosen for fresh transfer on day 5. A total of 441 embryos were intended to be frozen on day 3. Of those, 201 were not transferred nor vitrified on day 5–6 (45%), 3.35 embryos per patient. No significant difference was found between average day-3 rank of embryos ranked 1, 2 (3.12 vs 4.12, p = 0.074) and 3 (3.12 vs 4.08, p = 0.082) on day-5–6. To conclude, this study brings a different perspective to the comparison of day 3 and day 5 by following each embryo’s putative and actual designation. Day-3 ranking of embryo morphology did not provide a reliable prediction for blastocyst formation, transfer order and vitrification intention, and may support transfer or cryopreservation of blastocysts over cleavage stage embryos.

Metabolite supplementation during in vitro embryo development improves blastocyst quality, however, our understanding of the incorporation of metabolites during in vitro maturation (IVM) is limited. Two important metabolites, follistatin and choline, have beneficial impacts during in vitro culture; however, effects of supplementation during IVM are unknown. The objective of this study was to investigate combining choline and follistatin during IVM on bovine oocytes and subsequent early embryonic development. We hypothesized that supplementation of choline with follistatin would synergistically improve oocyte quality and subsequent early embryonic development. Small follicles were aspirated from slaughterhouse ovaries to obtain cumulus oocyte complexes for IVM with choline (0, 1.3 or 1.8 mM) and follistatin (0 or 10 ng/mL) supplementation in a 3 × 2 design. A subset of oocytes underwent transcriptomic analysis, the remaining oocytes were used for IVF and in vitro culture (IVC). Transcript abundance of CEPT1 tended to be reduced in oocytes supplemented with 1.8 mM choline and follistatin compared to control oocytes (P = 0.07). Combination of follistatin with 1.8 mM choline supplementation during maturation, tended (P = 0.08) to reduce CPEB4 in oocytes. In the blastocysts, HDCA8, NANOG, SAV1 and SOX2 were increased with choline 1.8 mM supplementation without follistatin (P < 0.05), while HDCA8 and SOX2 were increased when follistatin was incorporated (P < 0.05). The combination of choline and follistatin during oocyte maturation may provide a beneficial impact on early embryonic development. Further research is warranted to investigate the interaction between these two metabolites during early embryonic development and long-term influence on fetal development.

The Malabar dwarf puffer, Carinotetraodon travancoricus is the smallest known pufferfish (family Tetraodontidae) and one of the smallest freshwater fishes of the Indian subcontinent. Due to their miniature size, wacky behaviour and appearance, they are much preferred in the international aquarium fish trade, although little is known regarding their breeding activity in captivity and their embryonic development. The purpose of this study was to fill these knowledge gaps. Wild-caught Malabar dwarf puffers were acclimatised to conditions, and pairs were introduced to breeding tanks. Adult fishes were fed with live and frozen diets including Artemia nauplii, moina and bloodworm. During spawning seasons, adult fish displayed elaborate courtship behaviour around sunset. Carinotetraodon travancoricus is a batch spawner releasing 1 to 5 eggs per diem. The eggs were spherical, and non-sticky, with a diameter of 1.48 ± 0.1 mm, and hatching took place after 108 to 116 h post-incubation. The newly hatched larvae were 3.5 ± 0.2 mm in length, and weighed 2.9 ± 0.4 mg. The early larvae have substantial yolk and oil globules as an energy reserve. Histological studies on mature females suggested the batch spawning nature of the species and low fecundity. Given its unique reproductive behaviour and characters, in situ protected habitats are required to ensure their continued survival in the wild, apart from encouraging captive breeding to augment the demand in the international aquarium fish trade.

We have established trophoblast cell lines, from parthenogenesis-derived buffalo blastocysts. The buffalo trophoblast cells were cultured continuously over 200 days and 21 passages. These cells were observed by phase-contrast microscopy for their morphology and characterized by reverse transcriptase polymerase chain reaction and immunofluorescence against trophoblast-specific markers and cytoskeletal proteins. Trophoblast cells showed positive staining for CDX2, a marker of these cells at both blastocyst and cell line levels. Epithelial morphology of these cells was revealed by positive staining against cytokeratins and tubulin but not against vimentin and dolichos biflorus agglutinin. Gene expression profiles of many important placenta-specific genes were studied in the primary trophectoderm outgrowths, which were collected on days 0, 5, 9, 12 and 15 of culture and trophoblast cell line at passages 12–15. Therefore, the trophoblast cell line derived can potentially be used for in vitro studies on buffalo embryonic development.