Division of Histology and Cell Biology, Department of Anatomy, Jichi Medical University School of Medicine, Yakushiji 3311-1, Shimotsuke-shi, Tochigi 329-0498.
Acta Histochem Cytochem. 2010 May 1;43(2):83-8. doi: 10.1267/ahc.10001. Epub 2010 Apr 21.
Cadherins are a family of transmembrane glycoproteins that mediate cell-to-cell adhesion. A change in cadherin type in cells, i.e., cadherin switching, induces changes in the character of the cell. Recent studies of the developing rat adenohypophysis found that primordial cells co-expressed E- and N-cadherins, but that hormone-producing cells lost E-cadherin and ultimately possessed only N-cadherin. In the present study, we examined the roles of cadherin switching in cytogenesis of anterior pituitary cells by observing prolactin mRNA and protein expression in lactotrophs that were transformed with an E-cadherin expression vector. In hormone-producing cells that were transfected with a pIRES2-ZsGreen1 plasmid with a full-length E-cadherin cDNA (rE-cad-IZ) insert in primary culture, we detected E- and N-cadherins on plasma membrane and E-cadherin in cytoplasm. In these rE-cad-IZ-transfected cells, in situ hybridization revealed prolactin mRNA signals that were at a level identical to that in control cells, while prolactin protein was barely detectable using immunocytochemistry. The mean signal intensity of prolactin protein in rE-cad-IZ-transfected cells was approximately one fourth that in intact cells and in null-IZ-transfected cells (P<0.01). These results suggest that the expression of E-cadherin does not affect prolactin mRNA transcription; rather, it reduces prolactin protein content, presumably by affecting trafficking of secretory granules.
钙黏蛋白是一类介导细胞间黏附的跨膜糖蛋白。细胞中钙黏蛋白类型的改变,即钙黏蛋白转换,会诱导细胞特征的变化。最近对发育中的大鼠腺垂体的研究发现,原始细胞共同表达 E-和 N-钙黏蛋白,但产生激素的细胞失去了 E-钙黏蛋白,最终只拥有 N-钙黏蛋白。在本研究中,我们通过观察转染 E-钙黏蛋白表达载体的催乳素细胞中催乳素 mRNA 和蛋白的表达,来研究钙黏蛋白转换在前垂体细胞发生中的作用。在原代培养中,用含有全长 E-钙黏蛋白 cDNA(rE-cad-IZ)插入物的 pIRES2-ZsGreen1 质粒转染的激素产生细胞中,我们检测到质膜上的 E-和 N-钙黏蛋白以及细胞质中的 E-钙黏蛋白。在这些 rE-cad-IZ 转染的细胞中,原位杂交显示催乳素 mRNA 信号与对照细胞相同,但免疫细胞化学几乎检测不到催乳素蛋白。rE-cad-IZ 转染细胞中催乳素蛋白的平均信号强度约为完整细胞和空载体转染细胞的四分之一(P<0.01)。这些结果表明,E-钙黏蛋白的表达不会影响催乳素 mRNA 的转录;相反,它降低了催乳素蛋白的含量,可能是通过影响分泌颗粒的运输。