摘要
目的 :本文采用免疫组织化学方法观察Myb转录因子家族成员c myb蛋白在小鼠精子和卵母细胞 卵丘复合物中的分布。方法 :建立小鼠体外受精模型 ,用不同浓度反义c myb寡脱氧核苷酸 (c mybASODNs)与精子、卵母 卵丘细胞复合物共孵育观察其对小鼠体外受精率的影响 ,并进一步探讨c mybASODNs对小鼠体外受精率的影响机制。结果 :在颗粒细胞胞核和精子的头部有c myb蛋白分布。c mybASODNs呈剂量依赖性抑制小鼠体外受精率 (小鼠体外受精率在空白组、低、中、高浓度c mybASODNs组、无义tatODNs组分别为 34.97%、30 .89%、2 0 .14 %、16 .6 8%、34.4 7% )。GABA、P4、Verapamil和dbcAMP与c mybASODNs共同作用时 ,GABA、P4、dbcAMP三者均逆转c mybASODNs对受精的抑制作用 ,(小鼠体外受精率在空白组、中浓度c mybASODNs组、GABA、P4、Verapamil和dbcAMP组、分别为 34.81%、2 2 .96 %、4 0 .83%、39.12 %、7.4 6 3%、4 0 .6 1% ) ,GABA、P4、和dbcAMP三者对精子中的c myb蛋白免疫组化染色阳性的细胞百分率无明显影响。Verapamil能抑制小鼠体外受精 ,并协同c mybASODNs的抑制作用 ,且使精子中的c myb蛋白免疫组化染色阳性的细胞百分率明显下降。结论 :c mybA SODNs与受精密切相关 ,Verapamila可能通过调节精子中的c
Aim: To investigate the distribution of c-myb,an oncoprotein,in mouse oocytes-cumulus cell complex and sperm immunohistochemically. Methods: To study the effect of c-myb on mouse fertilization in vitro,various concentration of c-myb antisenseoligodeoxynucleotides (c-myb ASODNs) were incubated with sperms and oocytes during fertilization.To explore the possible mechanism involved in fertilization,the relationship between c-myb ASODNs and GABA or dbcAMP or Verapamil or Progesterone in ferti- lization was also observed by immunohistochemical methods. Results: c-myb oncoprotein was observed on the nucleus of cumulus cell and head of sperm. c-myb ASODNs inhibited the rate of fertilization in vitro in a dose-dependant way.The fertilization rates of the control group,low(5 μmol/L),medium(10 μmol/L),high(20 μmol/L) concentration c-myb ASODNs groups and nonsense tat oligodeoxynucleotides(20 μmol/L) group were 34.97%, 30.89%, 20.14%, 16.68%, 34.47%,respectively. All of GABA,Progesterone and dbcAMP inversed the c-myb ASODNs inhibition effects on fertilization rate,but neither of them showed significant effect on the percentages of immunohistochemical stain of Myb on sperm and cumulus cells.By contrast,Verapamil inhibited the fertilization rate.Co-treated with c-myb ASODNs, Verapamil showed synergic inhibiting effects on the fertilization with c-myb ASODNs.Verapamil also inhibited the expression of Myb on head of sperm. The fertilization rates of the control group,medium (10 μmol/L) concentration c-myb ASODNs group, GABA group, P 4 group, Verapamil group,dbcAMP group were 34.81%,22.96%,40.83%,39.12%,7.46%,40.61%,respectively. Conclusion: c-myb ASODNs is closely correlated with fertilization.Verapamil can inhibit fertilization in vitro through regulating Myb expression of sperm,while GABA, dbcAMP and Verapamil may affect the process of fertilization through the way other than Myb expression.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2005年第1期81-85,i003,共6页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助课题 (3 0 2 60 0 3 5 )
江西省自然科学基金项目资助项目 (9940 0 0 1)
