体外成熟液中添加PAF对牦牛卵母细胞发育能力及基因表达的影响

doi:10.11843/j.issn.0366-6964.2020.03.011

Abstract

Abstract: The objectives of this study were to evaluate the effects of platelet-activating factor (PAF) supplementation in in vitro maturation (IVM) medium on yak (Bos grunniens) oocyte development competence and gene expression. A total of 1 025 yak cumulus-oocyte complexes (COCs) were assigned to 4 groups. COCs in different groups were matured in IVM medium with different concentrations of PAF (0, 10^-8, 10^-7, and 10^-6mol·L^-1), followed by in vitro fertilization (IVF) with cattle sperm and in vitro culture (IVC). The maturation rate, cleavage rate and blastocyst rate of oocytes, and the relative expression level of pro-apoptosis (BAX), anti-apoptosis (BCL-2), epidermal growth factor (EGF) and its receptor (EGFR), transcription factors (OCT-4 and NANOG) and c-fos genes in COCs and embryos were compared among different groups. The results showed that oocytes in IVM medium with 10^-7mol·L^-1 PAF had higher maturation rate((92.16±0.19)%), cleavage rate((77.20±0.85)%) and blastocyst rate((46.71±0.68)%) than those in the other groups (P<0.05). The results of qPCR showed that the expression level of BCL-2, EGF, EGFR, c-fos were significantly up-regulated (P<0.05), but the expression level of pro-apoptotic gene (BAX) was significantly down-regulated in matured COCs (P<0.05) in 10^-7 mol·L^-1 PAF group. The expression level of EGF, EGFR and OCT-4 in blastocysts were significantly up-regulated in 10^-7 mol·L^-1 PAF group (P<0.05). The expression level of BCL-2, BAX and NANOG genes in 2-, 4-, 8-cell, morula and blastocyst stages were not different among different groups, but the expression of BCL-2 was down-regulated and BAX was up-regulated in morula and blastocyst. In conclusion, the optimal PAF concentration in IVM medium (10^-7mol·L^-1) could improve oocyte maturation and embryo development, and regulate the expression of apoptosis and proliferation related genes of the yak.

Key words: yak, platelet-activating factor, oocyte, in vitro fertilization, qPCR

CLC Number:

S823.85

WANG Qin, XIONG Yan, ZI Xiangdong. Effects of PAF Supplementation during in vitro Maturation Medium on Yak Oocyte Development Competence and Gene Expression[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 514-523.

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Effects of PAF Supplementation during in vitro Maturation Medium on Yak Oocyte Development Competence and Gene Expression

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