【摘要】 研究不同浓度生长激素释放肽(growth hormone releasing peptide, GHRL)对牛卵母细胞体外成熟和发育的影响。应用免疫荧光染色法检测GHRL及生长激素促分泌素受体(growth hormone secretagogue receptor, GHSR)在牛卵和卵丘细胞中的定位，添加0、10、50、100 ng/mL的GHRL对牛卵母细胞进行体外成熟24 h后，检测卵丘细胞扩散程度；GDF 9、BMP 15 mRNA的相对表达量；统计卵母细胞成熟及胚胎发育情况。结果表明，牛卵胞质区有GHRL的表达，且GHSR主要定位于细胞膜上。与对照组相比，添加GHRL 50 ng/mL组卵丘细胞扩散程度显著增加(P<0.05),GHRL 100 ng/mL组显著减少(P<0.01);添加50 ng/mL组的极体排出率显著提高(P<0.05),且BMP 15和GDF 9 mRNA的相对表达量均显著高于对照组(P<0.01);体外成熟16 h时，添加50 ng/mL组极体排出率显著高于对照组(P<0.01),体外受精后，GHRL 50 ng/mL组卵裂率相比对照组无显著差异(P>0.05);添加GHRL 50 ng/mL体外成熟24 h和16 h受精后的卵裂率无显著差异(P>0.05),但24 h囊胚率(63.24%)显著高于16 h(P<0.05)。在试验条件下，添加50 ng/mL可促进卵母细胞的成熟，但抑制了囊胚的形成。研究结果可为牛卵母细胞体外成熟、体外受精的应用提供理论依据。更多还原

【Abstract】 The aim of this experiment was to investigate the effects of GHRL on the maturation and development of bovine oocytes in vitro.Immunofluorescence staining was used to detect the localization of GHRL and receptor GHSR in bovine oocytes and cumulus cells.After adding 0,10,50,100 ng/mL of GHRL,bovine oocytes were matured for 24hin vitro,the diffusion degree of cumulus cells was detected.The relative expression levels of GDF-9and BMP 15mRNA,oocyte maturation and embryo development were analyzed.The results showed that GHRL was expressed in the cytoplasm of bovine oocytes,and GHSR was mainly located in the cell membrane.Compared with the control group,the degree of cumulus cell diffusion was significantly increased in GHRL 50ng/mL group(P<0.05),significantly decreased in GHRL 100ng/mL group(P<0.01).The polar body excretion rate in 50ng/mL group was significantly increased(P<0.05),and the relative expression levels of BMP 15and GDF 9mRNA were significantly higher than those in control group(P<0.01).After 16hof invitro maturation,the polar body excretion rate of the 50ng/mL group was significantly higher than that of the control group(P<0.01).After in vitro fertilization,the cleavage rate of the GHRL 50ng/mL group had no significant difference compared with the control group(P>0.05).There was no significant difference in cleavage rate between GHRL 50ng/mL supplementation at 24hand 16hafter fertilization(P>0.05),but the blastocyst rate at 24hwas significantly higher than that at 16h(P<0.05).Under the conditions of this experiment,the addition of 50ng/mL can promote the maturation of oocytes,but inhibit the formation of blastocysts.It can mature in vitro of bovine oocytes and lay a theoretical foundation for the application of in vitro fertilization.更多还原