Tubular morphogenesis and mesenchymal interactions affect renin expression and secretion in SIMS mouse submandibular cells
Tubular morphogenesis and mesenchymal interactions affect renin expression and secretion in SIMS mouse submandibular cells
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DOI：
10.1006/excr.1999.4404
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发表时间：
1999-04-10
影响因子：
3.7
通讯作者：
Rougeon, F
中科院分区：
文献类型：
--
作者：
Laoide, BM;Gastinne, I;Rougeon, F
我们之前已经在腺病毒5e1a启动子的控制下，使来自青春期雄性小鼠的小鼠下颚腺（SMG）导管上皮细胞系SIMS永生，该细胞系转SV40大T抗原。在这里，我们证明了细胞外环境不仅在指导细胞的形态发生行为方面的作用，而且在肾素表达和分泌方面也指导了细胞的功能分化。首先，我们测量了极化的SIMS细胞的肾素活性。肾素的分泌水平较低，从肾顶区和基底外侧区都有；机制似乎是直接的，因为没有发现肾素在细胞中被转酶。其次，我们研究了同型和异型间充质细胞与SIMS细胞的相互作用。我们发现，在I型胶原凝胶中，上皮-间充质共培养可导致SIMS细胞的分支管状形态形成，并分泌大量肾素，可能作为前体形式，原肾素进入管腔。第三，我们研究了基底膜对SIMS细胞形态和功能的影响，发现仅这种结构就足以使促肾素和活性肾素的表达和分泌。最后，我们通过瞬时转染实验确定了SIMS细胞可以表达雄激素调节基因。此外，在基质培养中，雄激素受体的表达似乎是诱导的，这表明SIMS细胞系将有助于进一步研究在雄性小鼠中观察到的smg特异性基因的高水平表达的分子基础。(C) 1999学术出版社。
We have previously immortalized a mouse submandibular gland (SMG) ductal epithelial cell line, SIMS, from pubertal male mice transgenic for the SV40 large T antigen under the control of the adenovirus 5 E1A promoter. Here we demonstrate the role of the extracellular environment in directing not only the morphogenetic behavior of the cells, but also their functional differentiation in terms of renin expression and secretion. First, we measured renin activity of polarized SIMS cells. Low levels of renin are secreted from both the apical and the basolateral domains; the mechanism appears to be direct as no renin was found to be transcytosed across the cell. Second, we studied homotypic and heterotypic mesenchymal cell interactions with SIMS cells. We found that epithelial-mesenchymal coculture in collagen I gels results in branching tubular morphogenesis of SIMS cells and that significant amounts of renin are secreted, probably into the lumen, as the precursor form, prorenin. Third, we investigated the effects of the basement membrane on SIMS cell morphology and function and found that this structure alone is sufficient to allow expression and secretion of both prorenin and active renin. Finally, we established that SIMS cells can express androgen-regulated genes in a transient transfection assay. In addition, in Matrigel cultures androgen receptor expression appears to be induced, suggesting that the SIMS cell line will be useful for further studies on the molecular basis of the observed high-level expression of SMG-specific genes in male mice. (C) 1999 Academic Press.