Division of Renal Diseases and Hypertension, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA.
Methods Mol Biol. 2021;2224:123-132. doi: 10.1007/978-1-0716-1008-4_9.
Proteinuria is a widely used marker of renal disease and is strongly associated with renal and cardiovascular outcomes. The molecular mechanisms underlying filtration of serum proteins through the glomerular filtration barrier (GFB) remain to be determined. Since the GFB is a complex structure, studies of albumin or IgG trafficking in cultured cells in vitro may not fully recapitulate these processes in vivo. In other epithelial cells including renal proximal tubular cells, the neonatal Fc receptor (FcRn) is required to divert albumin and IgG from the degradative pathway which allows these proteins to be recycled or transcytosed. To examine the role of podocyte FcRn in albumin and IgG trafficking in vivo, we detail the creation of a podocyte-specific FcRn knockout mouse and describe methods for examining intraglomerular detection of albumin and IgG in these mice.
蛋白尿是肾脏疾病的常用标志物,与肾脏和心血管结局密切相关。然而,血清蛋白通过肾小球滤过屏障(glomerular filtration barrier,GFB)滤过的分子机制仍有待确定。由于 GFB 是一种复杂的结构,体外培养细胞中白蛋白或 IgG 转运的研究可能无法完全重现体内这些过程。在其他上皮细胞,包括肾近端小管细胞中,需要新生 Fc 受体(neonatal Fc receptor,FcRn)将白蛋白和 IgG 从降解途径中转移出来,从而使这些蛋白质能够被回收或转运。为了研究足细胞 FcRn 在白蛋白和 IgG 体内转运中的作用,我们详细介绍了一种足细胞特异性 FcRn 敲除小鼠的构建,并描述了在这些小鼠中检查肾小球内白蛋白和 IgG 检测的方法。