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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。这一结论将为人类生育调控和不孕症的治疗提供新思路
Objective To determine the cellular localization and expression of androgen receptor (AR) and follicle stimulating hormone receptor (FSHR) in rat testes. METHODS: Digoxigenin-labeled cRNA probes were used to perform in situ hybridization on frozen sections of testes of adult rats. At the same time, seminiferous tubules of adult rats were divided into Ⅱ ~ Ⅵ, Ⅶ ~ Ⅷ, Ⅸ ~ Ⅻ, Ⅹ Ⅲ ~ Ⅰ four stages, the total RNA was extracted, α 32 P labeled cDNA probe hybridization spots observed AR and FSHR mRNA expression of cell location and phase-dependent changes. Using the image analysis system, the average luminance of the positive hybridization signal per unit area was quantitatively analyzed. Results The positive hybridization signal of AR mRNA was located in the supporting cells and interstitial cells, the strongest in Ⅶ ~ Ⅷ and the weakest in Ⅸ ~ Ⅰ. The positive signals of FSHR mRNA in supporting cells were the strongest in ⅩⅢ ~ Ⅰ and the weakest in Ⅶ ~ Ⅷ. . There was a significant difference between the stages (P <0.01). Conclusion The different rules of AR and FSHR-dependent expression suggest that T (testosterone) and FSH (follicle stimulating hormone) act on different stages of spermatogenesis, indicating that testosterone and follicle-stimulating hormone can regulate spermatogenesis in adult rats. This conclusion will provide new ideas for the regulation of human fertility and the treatment of infertility