Rastaldi Maria Pia, Armelloni Silvia, Berra Silvia, Li Min, Pesaresi Marzia, Poczewski Helga, Langer Brigitte, Kerjaschki Dontscho, Henger Anna, Blattner Simone Monika, Kretzler Matthias, Wanke Rudiger, D'Amico Giuseppe
Renal Immunopathology Laboratory, Associazione Nuova Nefrologia, c/o San Carlo Borromeo Hospital, Via Pio II, 20153 Milan, Italy.
Am J Pathol. 2003 Sep;163(3):889-99. doi: 10.1016/S0002-9440(10)63449-9.
Several recent studies have focused on similarities between glomerular podocytes and neurons because the two cells share a specialized cytoskeletal organization and several expression-restricted proteins, such as nephrin and synaptopodin. In neurons, the small guanosine triphosphatase Rab3A and its effector rabphilin-3A form a complex required for the correct docking of synaptic vesicles to their target membrane. Because rabphilin-3A binds in neurons to cytoskeletal proteins also important for podocyte homeostasis, and the complex rabphilin-3A-Rab3A has been demonstrated in neurons and neuroendocrine cells, the aim of our work was to investigate their possible expression and regulation in podocytes. Normal kidneys from mouse, rat, and human were studied by immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction to evaluate the expression of Rab3A and rabphilin-3A. Double-staining immunohistochemistry and immunogold electron microscopy were then used to precisely localize the two proteins at the cellular and subcellular levels. Rab-3A and rabphilin-3A regulations in disease were then analyzed in growth hormone-transgenic mice, a well established model of focal and segmental glomerulosclerosis, and in human biopsies from proteinuric patients. Our results demonstrated that rabphilin-3A and Rab3A are present in normal mouse, rat, and human kidneys, with an exclusively glomerular expression and a comma-like pattern of positivity along the glomerular capillary wall, suggestive for podocyte staining. Co-localization of both molecules with synaptopodin confirmed their presence in podocytes. By immunogold electron microscopy both proteins were found around vesicles contained in podocyte foot processes. Their expression was increased in growth hormone-transgenic mice compared to their wild-type counterpart, and in a subset of biopsies from proteinuric patients. Our data, demonstrating the presence of two synaptic proteins in podocytes, further supports similarities between cytoskeletal and vesicular organization of podocytes and neurons. The altered expression observed in mouse and human proteinuric diseases suggests a possible role for these molecules in glomerulopathies.
最近的几项研究聚焦于肾小球足细胞与神经元之间的相似性,因为这两种细胞具有特殊的细胞骨架组织以及几种表达受限的蛋白质,如nephrin和突触素蛋白。在神经元中,小GTP酶Rab3A及其效应蛋白rabphilin - 3A形成一个复合物,该复合物是突触小泡正确对接至其靶膜所必需的。由于rabphilin - 3A在神经元中与对足细胞稳态也很重要的细胞骨架蛋白结合,并且rabphilin - 3A - Rab3A复合物已在神经元和神经内分泌细胞中得到证实,我们研究的目的是调查它们在足细胞中可能的表达和调控情况。通过免疫组织化学、蛋白质印迹法和逆转录聚合酶链反应对小鼠、大鼠和人类的正常肾脏进行研究,以评估Rab3A和rabphilin - 3A的表达。然后使用双重染色免疫组织化学和免疫金电子显微镜在细胞和亚细胞水平精确地定位这两种蛋白质。随后在生长激素转基因小鼠(一种成熟的局灶节段性肾小球硬化模型)以及蛋白尿患者的人类活检组织中分析疾病状态下Rab - 3A和rabphilin - 3A的调控情况。我们的结果表明,rabphilin - 3A和Rab3A存在于正常小鼠、大鼠和人类肾脏中,仅在肾小球表达,并且沿着肾小球毛细血管壁呈逗号样阳性模式,提示足细胞染色。这两种分子与突触素蛋白的共定位证实了它们在足细胞中的存在。通过免疫金电子显微镜发现这两种蛋白质存在于足细胞足突内的小泡周围。与野生型对照相比,它们在生长激素转基因小鼠以及蛋白尿患者的一部分活检组织中的表达增加。我们的数据表明足细胞中存在两种突触蛋白,进一步支持了足细胞与神经元在细胞骨架和小泡组织方面的相似性。在小鼠和人类蛋白尿疾病中观察到的表达改变表明这些分子在肾小球病中可能发挥作用。