Cyr D G, Hermo L, Robaire B
Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec, Canada.
Endocrinology. 1993 Mar;132(3):1115-24. doi: 10.1210/endo.132.3.7679970.
The formation of junctional complexes between adjacent epithelial principal cells leads to formation of the blood-epididymal barrier; this barrier is complete by 21 days of postnatal age. Cadherins are cell surface proteins that mediate intercellular adhesion and are involved in the formation of adherence, gap, and tight junctions between epithelial cells. In the adult rat epididymis, epithelial cadherin (E-Cad) is localized in principal cells; E-Cad mRNA concentrations are androgen dependent in this tissue. The objectives of this study were to determine the regulation of E-Cad mRNA concentrations and the pattern of immunocytochemical localization of E-Cad during epididymal development. Using Northern blot analysis, we noted that in the caput-corpus epididymidis, there was a 3-fold increase in E-Cad mRNA concentrations between 7-14 days; an additional 3-fold increase between days 35-42, when E-Cad mRNA concentrations reached their peak, was noted. A dramatic decrease in E-Cad mRNA was observed between 42-49 days of age. This effect was transitory as E-Cad mRNA concentrations returned to almost 80% of peak concentrations on day 56 and remained constant thereafter. In the cauda epididymidis, E-Cad mRNA concentrations increased by only 1.6-fold between days 7-21. E-Cad mRNA concentrations then decreased by 70% to their lowest concentrations on day 56. There was a 2-fold increase in E-Cad mRNA concentrations between postnatal ages 56-91 days. These results suggest that the developmental regulation of E-Cad mRNA concentrations is segment specific. A subsequent study on the longitudinal distribution of E-Cad mRNA levels in six epididymal segments at 21, 42, and 56 days of age revealed that the relative proportion of E-Cad mRNA along the epididymis changes as a function of age. An immunocytochemical study with the light microscope, using an anti-E-Cad antibody, demonstrated that the localization and relative concentrations of E-Cad varied as a function of age. On day 15, the immunoperoxidase staining of the entire epididymal epithelium was apical, with the weakest staining in the cauda epididymidis. By day 21, the reaction spread to cover the supranuclear region of the principal cells in all segments, while on day 39, it covered the entire cytoplasm of these cells, suggesting a high rate of synthesis or storage of the protein. At later time intervals, the intensity of staining over the principal cells appeared to increase with age.(ABSTRACT TRUNCATED AT 400 WORDS)
相邻上皮主细胞之间连接复合体的形成导致血-附睾屏障的形成;该屏障在出生后21天发育完全。钙黏蛋白是介导细胞间黏附的细胞表面蛋白,参与上皮细胞间黏附连接、缝隙连接和紧密连接的形成。在成年大鼠附睾中,上皮钙黏蛋白(E-Cad)定位于主细胞;该组织中E-Cad mRNA浓度依赖雄激素。本研究的目的是确定附睾发育过程中E-Cad mRNA浓度的调节以及E-Cad免疫细胞化学定位模式。通过Northern印迹分析,我们注意到在附睾头-体部,E-Cad mRNA浓度在7至14天之间增加了3倍;在35至42天之间又增加了3倍,此时E-Cad mRNA浓度达到峰值。在42至49天龄时观察到E-Cad mRNA显著下降。这种效应是短暂的,因为E-Cad mRNA浓度在第56天恢复到峰值浓度的近80%,此后保持稳定。在附睾尾部,E-Cad mRNA浓度在7至21天之间仅增加了1.6倍。然后E-Cad mRNA浓度在第56天下降70%至最低浓度。在出生后56至91天之间,E-Cad mRNA浓度增加了2倍。这些结果表明,E-Cad mRNA浓度的发育调节具有节段特异性。随后一项关于21、42和56天龄时六个附睾节段E-Cad mRNA水平纵向分布的研究表明,沿附睾的E-Cad mRNA相对比例随年龄变化。一项使用抗E-Cad抗体的光镜免疫细胞化学研究表明,E-Cad的定位和相对浓度随年龄变化。在第15天,整个附睾上皮的免疫过氧化物酶染色位于顶端,在附睾尾部染色最弱。到第2天,反应扩散到覆盖所有节段主细胞的核上区域,而在第39天,它覆盖了这些细胞的整个细胞质,表明该蛋白的合成或储存率很高。在随后的时间段,主细胞上的染色强度似乎随年龄增加。(摘要截于400字)