次黄嘌呤对猪卵母细胞体外自发成熟抑制作用的研究

STUDIES ON THE INHIBITORY EFFECT OF HYPOXANTHINE ON THE SPONTANEOUS MEIOTIC MATURATION OF PORCINE OOCYTES IN VITRO

利用猪卵母细胞体外无血清培养技术,研究了次黄嘌呤(HX)对猪卵母细胞体外自发成熟的抑制作用。猪卵丘-卵母细胞复合体(COC)和裸卵母细胞(DO)取自初情期猪卵巢,培养在M-199培养液中,并施以各种处理。培养不同时间后,观察卵母细胞核成熟(GVBD)情况。实验结果表明:(1)HX(1-4mmol/L)对猪COC的自发成熟具有抑制作用,且具有剂量依赖关系。4mmol/L的HX对COC和DO自发成熟的抑制作用无明显差异(P>005);(2)HX对COC的自发成熟的抑制作用,随培养时间的延长而减弱;(3)COC在含有4mmol/LHX的M-199培养液中培养24h后转移到不含HX的培养液中继续培养时,仍能够自发成熟。在体外,HX能够可逆地部分抑制猪卵母细胞体外自发成熟,该作用具有剂量依赖性。

Using the serum-free culture technique for pig oocyte,the inhibitory effect of hypoxanthine (HX) on the spontaneous meiotic maturation of porcine oocytes was investigated.Pig cumulus oocyte complexes and denuded oocytes (COC and DO) were obtained from the ovaries of prepubertal gilts and cultured in medium 199.Different treatments were carried out on the oocytes during the culture.The incidence of germinal vesicle breakdown (GVBD) was examined at the end of the culture.The results showed:(1)HX,at the concentration of 1-4mmol/L,could significantly inhibit the spontaneous maturation of pig COC in a dose-dependent manner.The inhibitory effect of 4mmol/L HX on COC was not different from that on DO.(2)The inhibitory effect of HX (4mmol/L) on COC decreased in a time-dependent manner within the 72-hour culture period.(3)COC still could mature spontaneously in the following subsequent culture period when transferred from HX-supplemented medium to HX-free medium.These results indicated that HX could partially inhibit the spontaneous meiotic maturation of pig oocytes in vitro in a dose-dependent manner;This inhibitory effect is reversible.