Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, India.
Department of Veterinary Gynaecology and Obstetrics, GBPUA &T, Pantnagar, India.
Reprod Domest Anim. 2020 Oct;55(10):1362-1370. doi: 10.1111/rda.13782. Epub 2020 Aug 17.
In vitro-produced (IVP) embryos are reported to be developmentally lesser competent than in vivo-derived (IVD) embryos and supposed to differ in the expression of genes related with glucose metabolism. So, the present study was conducted to analyse the expression pattern of GLUT 1, 5, 8 and citrate synthase (CS) in oocytes and embryos produced in vivo or in vitro in buffalo. IVD embryos were obtained from 18 superovulated buffaloes. IVP embryos were obtained from slaughterhouse-derived oocytes subsequently subjected to in vitro fertilization and culture. Total RNA was isolated from different stages of oocytes (immature and in vitro matured) and embryos (8-16 cell to blastocysts of IVP embryos and morula to blastocysts of IVD embryos). Results demonstrated that the expression of GLUT1, GLUT 8 increased from 8 to 16 cells to blastocyst and was significantly (p < .05) higher in IVP embryos. Expression of both genes was (p < .05) higher in IVD than in IVP blastocysts; though GLUT5 transcripts were not detected at 8- to 16-cell stage IVP embryos, significantly (p < .05) higher transcripts were found at morula and blastocyst stages irrespective of embryo source with significantly (p < .05) higher expression in IVD embryos compared to IVP embryos. No significant difference was observed in citrate synthase expression in embryos at morula stage irrespective of the embryo source while significantly (p < .05) higher transcript level was observed at blastocyst stage with no difference between in vivo and in vitro embryos. It can be concluded that expression of GLUTs and CS is upregulated with progression of embryonic stage and expression is higher in in vivo embryos than in vitro counter parts; thus, it can be said that in vivo-produced embryos are metabolically superior to in vitro embryos.
体外生产（IVP）的胚胎被报道在发育能力上不如体内生产（IVD）的胚胎，并且在与葡萄糖代谢相关的基因表达上可能存在差异。因此，本研究旨在分析水牛体内和体外生产的卵母细胞和胚胎中 GLUT1、5、8 和柠檬酸合酶（CS）的表达模式。IVD 胚胎是从 18 头超排的水牛中获得的。IVP 胚胎是从屠宰场获得的卵母细胞进行体外受精和培养后获得的。从不同阶段的卵母细胞（不成熟和体外成熟）和胚胎（8-16 细胞至囊胚的 IVP 胚胎和桑椹胚至囊胚的 IVD 胚胎）中分离总 RNA。结果表明，GLUT1 和 GLUT8 的表达从 8-16 细胞期至囊胚期增加，并且在 IVP 胚胎中显著（p<.05）更高。这两个基因在 IVD 胚胎中的表达均显著（p<.05）高于 IVP 胚胎中的表达；尽管在 8-16 细胞期的 IVP 胚胎中未检测到 GLUT5 转录本，但在桑椹胚和囊胚期均发现显著（p<.05）更高的转录本，无论胚胎来源如何，IVD 胚胎中的表达均显著（p<.05）高于 IVP 胚胎。在胚胎的桑椹胚期，柠檬酸合酶的表达在胚胎来源上没有差异，而在囊胚期，转录本水平显著（p<.05）升高，体内和体外胚胎之间没有差异。可以得出结论，GLUTs 和 CS 的表达随着胚胎阶段的进展而上调，并且体内胚胎的表达高于体外胚胎；因此，可以说体内生产的胚胎在代谢上优于体外胚胎。
