Nephrin localizes to the slit pore of the glomerular epithelial cell -: Rapid Communication
Nephrin localizes to the slit pore of the glomerular epithelial cell -: Rapid Communication
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DOI：
10.1046/j.1523-1755.1999.00719.x
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发表时间：
1999-10-01
影响因子：
19.6
通讯作者：
Abrahamson, DR
中科院分区：
文献类型：
--
作者：
Holzman, LB;St John, PL;Abrahamson, DR
背景。认识到由 NPHS1 基因编码的去氧肾上腺素蛋白的突变会单独导致细胞改变，从而导致足突消失和肾病范围蛋白尿，这强调了该蛋白在调节肾小球滤过器完整性中发挥的关键作用。本文报道了研究小鼠去氧肾上腺素生物学所需试剂的开发，并描述了去氧肾上腺素蛋白的初步表征。方法。克隆并测序了包含全长小鼠去氧肾上腺素开放阅读框的 cDNA。开发了针对小鼠去氧肾上腺素细胞质结构域的免疫亲和纯化多克隆抗血清。结果。在小鼠肾小球提取物中鉴定出的去氧肾上腺素是一种表观分子质量为 185 kDa 的糖蛋白。通过间接免疫荧光显微镜和免疫金电子显微镜检测，去氧肾上腺素仅存在于内脏肾小球上皮细胞中，其靶向成熟足细胞足突的细胞间连接。在新生小鼠肾小球发育过程中，抗肾上腺素免疫定位于分化足细胞之间最早的狭缝孔区域，狭缝隔膜首先可见的部位。d结论。作为免疫球蛋白超家族的假定细胞粘附分子，去氧肾上腺素可能参与足细胞足突之间的细胞间相互作用，并且可能代表裂隙隔膜的组成部分。
Background. Recognition that mutation of the protein nephrin, encoded by the NPHS1 gene, singly results in the cellular alterations that result in foot process effacement, and nephrotic range proteinuria emphasizes the pivotal role that this protein plays in regulating glomerular filter integrity. This article reports the development of reagents necessary to study the biology of nephrin in mouse, and describes the initial characterization of the nephrin protein.Methods. A cDNA including the full-length mouse nephrin open reading frame was cloned and sequenced. Immune-affinity purified polyclonal antiserum directed against the cytoplasmic domain of mouse nephrin was developed.Results. Nephrin identified in mouse glomerular extract was found to be a glycoprotein with an apparent molecular mass of 185 kDa. As detected by indirect immunofluorescence microscopy and immunogold electron microscopy, nephrin was located only in visceral glomerular epithelial cells, where it was targeted to intercellular junctions of mature podocyte foot processes. In developing glomeruli of newborn mouse, antinephrin immunolocalized to the earliest slit pore regions between differentiating podocytes, sites where slit diaphragms first become visible.dConclusion. As a putative cell adhesion molecule of the immunoglobulin superfamily, nephrin likely participates in cell-cell interactions between podocyte foot processes and may represent a component of the slit diaphragm.