c-erbB_2与小鼠体外受精及作用机理研究  

EXPERIMENTAL STUDY OF c-erbB_2 ON THE FERTILIZATION IN MOUSE

作　　者：钟志胜[1] 吴凌峰[1] 郑月慧[1] 方廉[1] 

机构地区：[1]江西医学院生理教研室,江西南昌330006

出　　处：《中国应用生理学杂志》2003年第1期60-64,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金 ( 30 2 6 0 0 35) ;江西省自然科学基金项目 ( 994 0 0 0 0 1)资助

摘　　要：目的和方法 :用免疫组织化学的方法观察ErbB2 在小鼠睾丸、附睾、卵巢、输卵管、卵母细胞 -卵丘细胞复合物及精子细胞上的分布。用不同浓度反义c erbB2 寡脱氧核苷酸 (c erbB2 ASODNs)与精子和卵母细胞 -卵丘细胞复合物共孵育 ,观察其对小鼠体外受精率的影响 ,并进一步探讨其机制。结果 :在附睾的小管上皮细胞的管腔面 ,卵丘细胞和精子的细胞膜上有ErbB2 蛋白分布。c erbB2 ASODNs呈剂量依赖方式抑制小鼠体外受精率。小鼠体外受精率在空白对照组、低、中、高浓度c erbB2 ASODNs组、无义tatODNs组分别为 38.3 %、1 9.6 %、1 0 .7%、5 .0 %、33 .8% ,同时伴有精子细胞上ErbB2 免疫组化染色明显下降 ,中、高c erbB2 ASODNs浓度组卵丘细胞贴壁生长受抑制。GABA和db cAMP均能增加精子小鼠体外受精率 ,两者均部分逆转c erbB2 ASODNs对受精的抑制作用 ,但对精子细胞上ErbB2 免疫组化染色均无明显影响 ,而Verapamil能抑制小鼠体外受精率 ,并协同c erbB2 ASODNs的抑制作用 ,且使精子细胞膜上ErbB2 免疫组化染色明显下降。结论 :c erbB2 ASODNs与受精密切相关 ,阻断Ca2 + 内流后能通过抑制精子的c erbB2表达 ,从而抑制小鼠体外受精 ,而GABA、dbcAMP可能不是通过c erbB2Aim and Methods:The distribution of ErbB 2 in mouse testis, epididymidis, ovaries, oocyte cumulus cells complexes in oviducts and sperms was investigated immunohistochemically. To study the effect of c erbB 2 on mouse fertilization in vitro , various concentrations of c erbB 2 antisense oligonucleotides (c erbB 2 ASODNs) were incubated with sperms and oocyte cumulus cells complexes during fertilization in vitro . To explore possible mechanisms involved in fertilization, the relationship between c erbB 2 ASODNs and GABA,or dbcAMP,or verapamil during fertilization in vitro was also observed. Results: ErbB 2 oncoprotein was observed in epithelial cells in epididymis, sperms and cumulus cells. C erbB 2 ASODNs inhibited the rate of fertilization in vitro in a dose dependant way. The fertilization rate of the control group, low(5 μmol/L), medium(10 μmol/L),high(20 μmol/L) concentration c erbB 2 ASODNs group, and nonsense at oligonucleotides group (20 μmol/L) was 38.3%, 19.6%, 10.7%, 5.0%, and 33.8% respectively. Integral optical density immustaining of ErbB 2 in sperms was notably reduced. Medium and high concentration of c erbB 2 ASODNs notably inhibited cumulus cells adhering to inner wall of Petri dish.Treated alone with GABA or dbcAMP, the rate of fertilization was increased. Both GABA and dbcAMP partially inversed the ASODNs inhibition effect on fertilization rate, but neither of them showed significant effect on sperm integral optical density of ErbB 2 immustaining. In contrast, verapamil inhibited fertilization rate. Co treated with c erbB 2 ASODN, verapamil showed synergic inhibiting effect on fertilization with c erbB 2 ASODN. Verapamil also inhibited the expression of c erbB 2 in sperms. Conclusion:It is suggested that c erbB 2 is closely correlated with fertilization. Ca 2+ may inhibit fertilization in vitro through regulation the expression of c erbB 2 gene in sperm cells, while both of GABA and dbcAMP may affect the process of fertilizatio

关 键 词：C-ERBB2 反义寡脱氧核苷酸 体外受精 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]