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兔蛋白激酶C ζ在肾内的分布

Intrarenal distribution of rabbit PKC zeta.

作者信息

Hao C M, Breyer R M, Davis L S, Breyer M D

机构信息

Division of Nephrology, Veterans Administration Medical Center, Nashville, Tennessee, USA.

出版信息

Kidney Int. 1997 Jun;51(6):1831-7. doi: 10.1038/ki.1997.250.

Abstract

To elucidate roles of protein kinase C (PKC) zeta in rabbit kidney, PKC zeta was cloned from a rabbit kidney cortex cDNA library. Sequencing revealed a 2113 m insert with an open reading frame encoding a protein of 591 amino acids. The predicted amino acid sequence is 93.7% identical with rat PKC zeta. In situ hybridization in rabbit kidney with a riboprobe generated from the cloned cDNA, showed PKC zeta mRNA is highly expressed in proximal tubule, thick limb, and collecting duct. No message was detected over glomerular cells. Immunohistochemical studies using a monoclonal antibody against PKC zeta confirmed this distribution with low expression in vascular elements and high expression in tubule epithelium. Confocal microscopy showed diffuse cytosolic immunoreactivity in confluent cultured cortical collecting ducts (CCDs). However, in subconfluent cells, immunoreactivity was restricted to the peri-nuclear area. This differential distribution of PKC zeta in the CCD suggests that PKC zeta action be involved in growth and differentiation of the collecting duct. In conclusion, PKC zeta is differentially expressed in the rabbit kidney with high expression in the tubule epithelium and little expression in vascular elements. These studies suggest an important role for PKC zeta along the nephron.

摘要

为阐明蛋白激酶C(PKC)ζ在兔肾中的作用,从兔肾皮质cDNA文库中克隆了PKCζ。测序显示有一个2113 m的插入片段,带有一个编码591个氨基酸的蛋白质的开放阅读框。预测的氨基酸序列与大鼠PKCζ的同源性为93.7%。用克隆的cDNA产生的核糖探针在兔肾中进行原位杂交,结果显示PKCζ mRNA在近端小管、髓袢升支粗段和集合管中高度表达。在肾小球细胞上未检测到信号。使用抗PKCζ单克隆抗体进行的免疫组织化学研究证实了这种分布,在血管成分中表达较低,在肾小管上皮中表达较高。共聚焦显微镜显示,在汇合培养的皮质集合管(CCD)中有弥漫性的胞质免疫反应性。然而,在亚汇合细胞中,免疫反应性局限于核周区域。PKCζ在CCD中的这种差异分布表明,PKCζ的作用参与了集合管的生长和分化。总之,PKCζ在兔肾中差异表达,在肾小管上皮中高表达,在血管成分中低表达。这些研究表明PKCζ在肾单位中起重要作用。

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