促甲状腺激素释放激素受体-1(TRH-R1)蛋白在大鼠出生后睾丸不同发育阶段的表达和定位

目的研究促甲状腺激素释放激素受体-1(thyrotrophin-releasing hormone receptor type-1, TRH-R1)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TRH-R1在8d、15d、20d、35d、60d和90d大鼠睾丸中的表达和定位,并结合图像分析技术对免疫组化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-R1蛋白表达于15d以后各阶段的大鼠睾丸;而运用免疫组化在第8d即检测到TRH-R1的表达,以后发育过程中的各个阶段均有阳性反应细胞, TRH-R1定位于大鼠睾丸的间质细胞;免疫反应阳性物均位于胞膜和胞质,胞核区为阴性;图像分析结果表明,随着大鼠睾丸的发育,TRH-R1表达量呈增多趋势,且具有统计学差异(P＜0.01).结论本实验证明TRH-R1在出生后8d大鼠的睾丸内即有表达,并持续表达于其后各个发育阶段;TRH-R1定位于睾丸的间质细胞,其表达量随着增龄变化呈增多趋势,即同发育过程相关.

EXPRESSION AND LOCALIZATION OF TRH-R1 PROTEIN IN THE RAT TESTIS DURING POSTNATAL DEVELOPMENT

Objective To investigate theexpression of thyrotrophin-releasing hormone receptor type-1 (TRH-R1) in rat testis during postnatal development, and to explore its function in the regulation of reproduction development. Methods Western blotting and the immunohistochemical ABC method was used to detect the expression and localization of TRH-R1 in rat testes 8, 15, 20, 35, 60 and 90 days after birth. And the positive results of immunohitochemistry were analyzed with the image analysis system andstatistical method. Results Western blot showed that TRH-R1 was expressed in the testis of rats of 15 days and above. Butwith the immunohistochemical ABC method, we found that TRH-R1 appeared in 8-day rats. In all the following stages we also detected the immunochemical positive cells in testes. TRH-R1 was localizedin the interstitial cells of rattestisand positive substance was expressed in the membrane and cytoplasmof Leydig cells. Image analysis and statisticalinvestigation revealed that with the postnatal development of rats the intensity of TRH-R1 expression increased progressively. Conclusion TRH-R1 is expressed in rat testis during its postnatal development and localized in the Leydig cells, and such expression is development-dependent.