Coexpression of neuropilin-1, Flk1, and VEGF164 in developing and mature mouse kidney glomeruli
Coexpression of neuropilin-1, Flk1, and VEGF164 in developing and mature mouse kidney glomeruli
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DOI：
10.1152/ajprenal.2000.279.2.f275
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发表时间：
2000-08-01
影响因子：
4.2
通讯作者：
Abrahamson, DR
中科院分区：
文献类型：
--
作者：
Robert, B;Zhao, XM;Abrahamson, DR
神经纤毛蛋白-1是一种神经元细胞表面信号蛋白III受体蛋白，其对于发育中的外周神经系统传出神经的轴突引导是重要的，也已被鉴定为内皮细胞上的血管内皮生长因子（VEGF）受体。为了评估其在肾脏中的表达，我们对新生儿和成人肾脏总RNA进行了RT-PCR。一个403 bp的产物，这是预测从neuropilin-1 mRNA，被发现在这两个样品。核苷酸测序证实这些产物编码神经纤毛蛋白-1。新生儿和成人肾脏RNA的北方分析显示与大小合适的条带（约6 kb）特异性杂交。与小鼠特异性反义神经毡蛋白-1（35）S-cRNA探针的原位杂交显示新生儿和成人切片上不同的肾小球定位。在用针对另一种VEGF受体Flk1的反义cRNA探针和VEGF探针处理的切片中观察到类似的杂交模式。然而，VEGF杂交信号显着低于成人肾小球比神经纤毛蛋白-1和Flk1。由于neuropilin-1特异性结合VEGF（165）在人类中，我们进行了RT-PCR小鼠肾脏RNA的引物扩增的三个可变剪接的VEGF mRNA的亚型。我们的分析表明，在新生儿和成人肾脏中，VEGF mRNA的相对丰度为VEGF（164）>> VEGF（120）> VEGF（188）。我们的结论是，神经纤毛蛋白-1的表达，与Flk1和VEGF（164），共同有助于肾小球毛细血管的发展和维护。
Neuropilin-1, a neuronal cell surface semaphorin III receptor protein important for axonal guidance in developing peripheral nervous system efferents, has also been identified as a vascular endothelial growth factor (VEGF) receptor on endothelial cells. To evaluate its expression in kidney, we carried out RT-PCR on newborn and adult total renal RNAs. A 403-bp product, which was predicted to be that from neuropilin-1 mRNA, was found in both samples. Nucleotide sequencing confirmed that these products encoded neuropilin-1. Northern analysis of newborn and adult kidney RNA showed specific hybridization to appropriately sized bands of similar to 6 kb. In situ hybridization with a mouse-specific antisense neuropilin-1 (35)S-cRNA probe showed distinct glomerular localization on sections from both newborns and adults. Similar patterns of hybridization were seen in sections treated with antisense cRNA probes against another VEGF receptor, Flk1, and with VEGF probes. However, the VEGF hybridization signal was markedly less in adult glomeruli than those for neuropilin-1 and Flk1. Because neuropilin-1 specifically binds VEGF(165) in humans, we carried out RT-PCR on mouse kidney RNA with primers that amplified the three alternatively spliced isoforms of VEGF mRNA. Our analysis showed that for both newborn and adult kidneys, the relative abundance of VEGF mRNA was VEGF(164) >> VEGF(120) > VEGF(188). We conclude that the expression of neuropilin-1, in conjunction with Flk1 and VEGF(164), jointly contributes to the development and maintenance of glomerular capillaries.