生殖细胞核因子在不同成长期大鼠附睾中的表达

Expression of germ cell nuclear factor in rat epididymis during postnatal development

目的 探讨生殖细胞核因子 (GCNF)在大鼠附睾中的时空表达与分布。方法 应用免疫组化和蛋白质免疫印迹技术 ,检测GCNF蛋白在出生后 7、14、2 8、4 5、6 0、90天龄和 18月龄大鼠附睾内的表达、定位和发育变化 ,并通过凝胶成像及图像分析系统对实验结果进行统计学分析。结果 GCNF表达于出生后 14天龄至 18月龄大鼠附睾上皮的细胞核内 ,其表达呈明为的年龄相关性和附睾区段以及细胞特异性。 14天龄附睾上皮内出现散在阳性细胞核 ,2 8天龄时阳性细胞数和阳性强度明显增强 ,4 5天龄时达到高峰。成年后阳性细胞数维持不变而阳性染色强度开始逐渐减弱 ,18月龄时减弱非常明显。附睾管上皮内的各种细胞如主细胞、基细胞、顶细胞、狭窄细胞和亮细胞等均呈GCNF阳性染色 ,从附睾起始段到尾段到尾段均有阳性染色 ,其中以近端头部最强 ;远端尾部主细胞核阳性减弱非常明显甚至呈阴性 ,而亮细胞核则阳性较强。 14d后各年龄组附睾抽提物中均可见 5 50 0 0处有GCNF特异性条带 ,其年龄变化与免疫组化结果一致。结论 GCNF在大鼠附睾上皮内的表达与附睾上皮分化和发育过程明显相关 ,且其分布表现出明显的附睾区段和细胞特异性。

Objective To explore the expression and distribution of germ cell nuclear factor (GCNF),an orphan receptor of the nuclear receptor superfamily which is supposed to have multifunction including spermatogenesis,neurogenesis and embryo development,in adult rat epididymis and young rats during postnatal development.Methods Whole epididymis was collected from SD rats' aged 7,14,28,45,60 and 90 days,and 18 months.The protein expression of GCNF during rat epididymal development and its cellular localizations were examined by Western blotting and immunohistochemnistry respectively. Results GCNF was immunolocalized in the nuclei of epididymal epithelial cells with negatively stained cytoplasm in rats during postnatal development.Study of postnatal development revealed that the density of GCNF protein (about 55 000) was age-dependent.GCNF protein was first detected in the rats aged day 14.The number and intensity of positive cells increased along with differentiation and proliferation of the epithelium,and peaked when the rats were aged 45 days and then decreased gradually.The results of Western-blotting confirmed those of immuneohistochemistry.In adult rats,the GCNF immunoreactive particles were observed in all cell types of epididymis,namely,principal cell,basal cell,apical cell,narrow cell,and clear cell and halo cell.GCNF in the principal cells exhibited a region-spectific and cell-specific expression pattern that was observed from the age of 14 days.The GCNF expression was gradually decreased from the initial segment to proximal cauda epididymis.There was a predominant expression in the proximal caput region.The principal cells in the same segment also displayed a variable staining pattern,with the nucleus being intensely,moderately,or weekly reactive.In the distal cauda epididymis,only the nuclei of clear cells displayed an intensive staining,as compared with very weak staining in the principal.Conclusion The expression of GCNF may be concerned with the differentiation and development of epididymal epithelium,and its expression pattern is epididymal segment-specific and cell-specific.