Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines
Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines
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DOI：
10.1006/excr.1997.3739
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发表时间：
1997-10-10
影响因子：
3.7
通讯作者：
Zeller, R
中科院分区：
文献类型：
--
作者：
Mundel, P;Reiser, J;Zeller, R
成熟的足细胞是最复杂的分化细胞之一，具有高度分叉的足突，对肾脏的肾小球滤过至关重要。这种分化的足细胞不能复制，培养初级足细胞会导致快速生长停滞。因此，建立了条件永生化的小鼠足细胞克隆(MPC)，该克隆在允许的条件下培养具有高度的增殖性。不允许的条件使大多数MPC细胞在6天内停止生长，并诱导分化的足细胞的许多特征。增殖和分化的MPC细胞都表达WT-1蛋白，在分化过程中有序排列的肌动蛋白纤维和微管延伸到形成细胞的过程中，这使人想起体内的足细胞过程。这些细胞骨架重排和突起的形成伴随着突触素合成的开始，突触素是一种与肌动蛋白相关的蛋白质，标志着特定分化的足细胞。此外，在分化的MPC细胞中，焦点接触被重新排列成有序的图案。最重要的是，电生理学研究表明，分化的MPC细胞通过改变细胞内钙离子浓度来对血管活性多肽缓激肽做出反应。这些结果提示足细胞在肾小球滤过中的调节作用。综上所述，这些研究证实，条件永生化的MPC细胞在体内保持着类似于足细胞的分化潜力。因此，首次利用体外诱导模型研究了足细胞分化和突起形成的决定步骤。(C)1997年学术出版社。
Mature podocytes are among the most complex differentiated cells and possess a highly branched array of foot processes that are essential to glomerular filtration in the kidney. Such differentiated podocytes are unable to replicate and culturing of primary podocytes results in rapid growth arrest. Therefore, conditionally immortalized mouse podocyte clones (MPC) were established, which are highly proliferative when cultured under permissive conditions. Nonpermissive conditions render the majority of MPC cells growth arrested within 6 days and induce many characteristics of differentiated podocytes. Both proliferating and differentiating MPC cells express the WT-1 protein and an ordered array of actin fibers and microtubules extends into the forming cellular processes during differentiation, reminiscent of podocyte processes in vivo. These cytoskeletal rearrangements and process formation are accompanied by the onset of synaptopodin synthesis, an actin-associated protein marking specifically differentiated podocytes. In addition, focal contacts are rearranged into an ordered pattern in differentiating MPC cells. Most importantly, electrophysiological studies demonstrate that differentiated MPC cells respond to the vasoactive peptide bradykinin by changes in intracellular calcium concentration. These results suggest a regulatory role of podocytes in glomerular filtration. Taken together, these studies establish that conditionally immortalized MPC cells retain a differentiation potential similar to podocytes in vivo. Therefore, the determinative steps of podocyte differentiation and process formation are studied for the first time using an inducible in vitro model. (C) 1997 Academic Press.