雄激素受体和卵泡刺激素受体在成年大鼠睾丸中的期依赖性表达

STAGE-DEPENDENT EXPRESSION OF ANDROGEN RECEPTOR AND FSH RECEPTOR IN ADULT RAT TESTIS

目的 确定雄激素受体 (AR)和卵泡刺激素受体 (FSHR)在大鼠睾丸中的细胞定位和表达变化。 方法 利用地高辛标记的cRNA探针 ,在成年大鼠睾丸冰冻切片上进行原位杂交 ;同时利用透光显微切割技术将成年大鼠曲细精管分为Ⅱ～Ⅵ、Ⅶ～Ⅷ、Ⅸ～Ⅻ、ⅩⅢ～Ⅰ 4个阶段 ,提取总RNA ,用α 32 P标记的cDNA探针进行斑点杂交 ,观察AR和FSHRmRNA表达的细胞定位和期依赖性变化。利用图像分析系统 ,对阳性杂交信号单位面积平均辉度进行定量分析。 结果 ARmRNA阳性杂交信号位于支持细胞和间质细胞 ,于Ⅶ～Ⅷ期最强 ,Ⅸ～Ⅰ期最弱 ;FSHRmRNA阳性杂交信号位于支持细胞 ,于ⅩⅢ～Ⅰ期最强 ,Ⅶ～Ⅷ期最弱。各阶段之间具有非常显著性差异 (P<0 0 1)。 结论 AR和FSHR期依赖性表达的不同规律提示T(睾酮 )和FSH(卵泡刺激素 )作用于精子发生的不同阶段 ,说明睾酮和卵泡刺激素协同作用调节成年大鼠的精子发生。

Objective To determine the cellular localization and expression pattern of AR and FSHR in adult rat testis must be helpful to understand the action site and mechanisms that T and FSH regulate spermatogenesis. Methods We applied in situ Hybridization to detect the expression of AR and FSHR on adult testis, in which Dig labeled cRNA probe was used to carry out the experiment on frozen sections; at the same time, following the technique of transillumination assisted microdissection we separated seminiferous epithelium into four stages(Ⅱ Ⅵ,Ⅶ Ⅷ,Ⅸ Ⅻ and ⅩⅢ Ⅰ), extracted total RNA and carried out dot hybridization, using α 32 P labeled cDNA probe, in order to test qualitatively and quantitatively the location of AR and FSHR mRNA and their expression pattern in adult rat testis. Results Our results showed that the positive signal of AR mRNA was located in Sertoli cells and Leydig cells. The signal in Sertoli cells began to appear in Ⅱ Ⅵ stages, strongest in Ⅶ Ⅷ stages and weakest in Ⅸ Ⅰ stages ( P <0 01). The positive signal of FSHR mRNA was located in Sertoli cells strongest in ⅩⅢ Ⅰ stages, weakest in Ⅶ Ⅷ and intermediate in the other stages ( P <0 01). The different pattern of stage specific expression of AR and FSHR in adult rat testis suggested that T and FSH act on different stages during spermatogenesis. Conclusion We suppose that there is a coaction at regulation spermatogenesis between T and FSH. From this point of view, it may provide new ideas for designing contraceptive strategies and treating human infertility.