Panda Bibhudatta S K, Pandey Sriti, Somal Anjali, Parmar Mehtab S, Bhat Irfan A, Baiju Indu, Bharti Mukesh K, Sai Kumar G, Chandra Vikash, Sharma G Taru
Reproductive Physiology Laboratory, Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, Bareilly, U.P., India.
Division of Veterinary Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, Bareilly, U.P., India.
Theriogenology. 2017 Aug;98:116-122. doi: 10.1016/j.theriogenology.2017.05.008. Epub 2017 May 7.
The aim of the present study was to determine potential role of leptin on in vitro developmental competence of buffalo oocytes and embryos. Slaughterhouse derived culture grade buffalo cumulus oocyte complexes (COCs) were matured in vitro (IVM) with leptin (10 ng/ml) or without leptin (control). In each experiment, a pool of matured COCs was used for further in vitro embryo production and another pool of COCs was used for cumulus cells and mature oocytes isolation to study the relative mRNA expression of developmentally important genes. Presumptive zygotes were cultured in embryo culture (IVC) media supplemented with leptin (10 ng/ml) or without leptin (control). Cleavage rate was higher (p < 0.05) when leptin was supplemented during IVM + IVC, both, as compared to other groups. Higher cleavage rate was observed in leptin-treated groups, though it was non-significant. Blastocyst rate was higher (p < 0.05) in all the leptin treated groups. The relative mRNA expression of LEPR (Ob-Rb), HAS2 and EGFR was significantly (p < 0.05) up-regulated and the expression of CASPASE3 was down-regulated in cumulus cells of leptin-treated groups. The expression of GDF9, BMP15, GLUT1, LEPR and CASPASE3 transcripts in leptin and non-treated oocytes did not differ. The relative mRNA expression of POU5F1and LEPR transcripts in blastocysts was higher (p < 0.05) in leptin-treated groups; the change in expression of GLUT1, INF-τ and CASPASE3 transcripts was not significant (p > 0.05). Thus, it is concluded that leptin promotes developmental competence of bubaline oocytes by modulating cumulus enabling factors and genes regulating pluripotentcy in the blastocysts.
本研究的目的是确定瘦素对水牛卵母细胞和胚胎体外发育能力的潜在作用。从屠宰场获取的培养级水牛卵丘卵母细胞复合体（COCs）在体外成熟（IVM）时，一组添加瘦素（10 ng/ml），另一组不添加瘦素（对照）。在每个实验中，一组成熟的COCs用于进一步的体外胚胎生产，另一组COCs用于分离卵丘细胞和成熟卵母细胞，以研究发育重要基因的相对mRNA表达。推测的合子在添加瘦素（10 ng/ml）或不添加瘦素（对照）的胚胎培养液（IVC）中培养。与其他组相比，在IVM + IVC过程中添加瘦素时，卵裂率更高（p < 0.05）。在瘦素处理组中观察到较高的卵裂率，尽管不显著。所有瘦素处理组的囊胚率均较高（p < 0.05）。在瘦素处理组的卵丘细胞中，LEPR（Ob-Rb）、HAS2和EGFR的相对mRNA表达显著上调（p < 0.05），而CASPASE3的表达下调。瘦素处理组和未处理组的卵母细胞中GDF9、BMP15、GLUT1、LEPR和CASPASE3转录本的表达没有差异。瘦素处理组囊胚中POU5F1和LEPR转录本的相对mRNA表达较高（p < 0.05）；GLUT1、INF-τ和CASPASE3转录本表达的变化不显著（p > 0.05）。因此，得出结论，瘦素通过调节卵丘促成因子和调控囊胚多能性的基因来促进水牛卵母细胞的发育能力。
