The role of Notch signaling in specification of podocyte and proximal tubules within the developing mouse kidney
The role of Notch signaling in specification of podocyte and proximal tubules within the developing mouse kidney
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DOI：
10.1111/j.1523-1755.2005.00627.x
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发表时间：
2005-11-01
影响因子：
19.6
通讯作者：
Kopan, R
中科院分区：
文献类型：
--
作者：
Cheng, HT;Kopan, R
Notch基因编码跨膜受体，这些受体通过与相邻细胞上的配体结合来介导细胞间相互作用。由于一些Notch通路基因缺陷的小鼠在胚胎早期死亡，Notch信号在肾脏发育中的作用尚未明确。我们使用一种针对γ-分泌酶切割的Notch 1的N末端的特异性抗体，在逗号形和S形小体中发现了Notch 1激活的证据。因此，我们在γ -分泌酶抑制剂N - S -苯基 - 甘氨酸 - 叔丁酯（DAPT）存在的情况下培养胚胎（E）第12.5天的小鼠后肾，以阻断Notch信号。观察到的肾上皮结构减少，近端小管和肾小球足细胞严重缺失。远端小管存在，但数量减少，并且伴有中间非上皮细胞的增加。通过培养第14.5天的后肾，我们观察到在DAPT处理3天后足细胞簇的形成。这些观察结果表明，γ -分泌酶活性（可能通过激活Notch）对于S形小体阶段之后的足细胞形成不是必需的，但在S形小体形成时对于近端小管和足细胞的命运是必需的。
Notch genes encode transmembrane receptors that mediate intercellular interaction by binding to the ligands on the adjacent cells. Due to early embryonic lethality in mice deficient for some Notch pathway genes, the role of Notch signaling for kidney development has not yet been defined. Using an antibody specific to the N-terminal end of gamma-secretase-cleaved Notch 1, we found evidence for Notch 1 activation in the comma-shaped and S-shaped bodies. We therefore cultured embryonic (E) day E12.5 mouse metanephroi in the presence of a gamma-secretase inhibitor, N-S-phenyl-glycine-t-butyl ester (DAPT), to block Notch signaling. Fewer renal epithelial structures were observed, with a severe deficiency in proximal tubules and glomerular podocytes. Distal tubules were present but at a reduced number, and this was accompanied by an increase in intervening, nonepithelial cells. By culturing day E14.5 metanephroi, we observed the formation of podocyte clusters after 3 days of DAPT treatment. These observations suggest that gamma-secretase activity, probably through activation of Notch, is not essential for podocyte formation beyond the stage of S-shaped body but is required for the proximal tubule and podocyte fates when S-shaped bodies are forming.