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目的研究促甲状腺激素释放激素受体-2(TRH-R2)在大鼠睾丸出生后不同发育阶段的表达,探讨其在生殖发育调节中的作用。方法应用蛋白质免疫印迹杂交技术以及免疫组织化学ABC法检测8d、15d、20d、35d、60d和90d大鼠睾丸中TRH-R2的表达和定位,并结合图像分析技术对免疫组织化学结果进行统计学分析,观察其在发育过程中的变化。结果免疫印迹杂交发现TRH-R2蛋白表达于出生后15d以后各阶段的大鼠睾丸;而免疫组织化学在第8d以后的各个发育阶段均检测到TRH-R2的表达,TRH-R2定位于大鼠睾丸的间质细胞,TRH-R2免疫反应阳性物位于细胞膜和细胞质内,细胞核为阴性;图像分析结果显示随着大鼠睾丸的发育,TRH-R2表达量逐渐增多,35d达到最大值,此后维持稳定水平,表达量在35d同其他阶段之间具有统计学差异(P<0·01)。结论TRH-R2在大鼠睾丸发育过程中均有表达,定位于睾丸的间质细胞,其表达量随着增龄变化而变化,即同发育过程相关。
Objective To investigate the expression of thyrotropin-releasing hormone receptor-2 (TRH-R2) in different developmental stages after birth in rats and to explore its role in the regulation of reproductive development. Methods The expression and localization of TRH-R2 in the testes of rats at 8d, 15d, 20d, 35d, 60d and 90d were detected by Western blotting and immunohistochemical ABC method, and the immunohistochemical results were analyzed by image analysis Analysis, observation of its changes in the development process. Results The expression of TRH-R2 protein was detected in all the stages of postnatal 15 days after immunoblotting. The expression of TRH-R2 was detected in all developmental stages after 8th day by immunohistochemistry. TRH-R2 was localized in rat Testis stromal cells, TRH-R2 immunoreactivity positive located in the cell membrane and cytoplasm, the nucleus was negative; image analysis showed that with the development of rat testes, TRH-R2 expression gradually increased, reached the maximum 35d, then maintained Stable level, the expression of 35d with other stages with a statistically significant difference (P <0.01). Conclusion TRH-R2 is expressed in rat testis and located in the interstitial cells of testis. The expression of TRH-R2 changes with aging, which is related to development.