Liu Yan, Pathak Narendra, Kramer-Zucker Albrecht, Drummond Iain A
Nephrology Division, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
Development. 2007 Mar;134(6):1111-22. doi: 10.1242/dev.02806. Epub 2007 Feb 7.
Epithelial tubules consist of multiple cell types that are specialized for specific aspects of organ function. In the zebrafish pronephros, multiciliated cells (MCCs) are specialized for fluid propulsion, whereas transporting epithelial cells recover filtered-blood solutes. These cell types are distributed in a ;salt-and-pepper' fashion in the pronephros, suggesting that a lateral inhibition mechanism may play a role in their differentiation. We find that the Notch ligand Jagged 2 is expressed in MCCs and that notch3 is expressed in pronephric epithelial cells. Morpholino knockdown of either jagged 2 or notch3, or mutation in mind bomb (in which Notch signaling is impaired), dramatically expands ciliogenic gene expression, whereas ion transporter expression is lost, indicating that pronephric cells are transfated to MCCs. Conversely, ectopic expression of the Notch1a intracellular domain represses MCC differentiation. Gamma-secretase inhibition using DAPT demonstrated a requirement for Notch signaling early in pronephric development, before the pattern of MCC differentiation is apparent. Strikingly, we find that jagged 2 knockdown generates extra cilia and is sufficient to rescue the kidney cilia mutant double bubble. Our results indicate that Jagged 2/Notch signaling modulates the number of multiciliated versus transporting epithelial cells in the pronephros by way of a genetic pathway involving repression of rfx2, a key transcriptional regulator of the ciliogenesis program.
上皮小管由多种细胞类型组成,这些细胞类型专门负责器官功能的特定方面。在斑马鱼前肾中,多纤毛细胞(MCCs)专门负责液体推进,而转运上皮细胞则回收滤过血液中的溶质。这些细胞类型在前肾中以“椒盐”方式分布,这表明侧向抑制机制可能在它们的分化中起作用。我们发现Notch配体Jagged 2在MCCs中表达,而notch3在前肾上皮细胞中表达。对Jagged 2或notch3进行吗啉代敲低,或在mind bomb(其中Notch信号传导受损)中发生突变,会显著扩大纤毛生成基因的表达,而离子转运蛋白的表达则丧失,这表明前肾细胞转变为MCCs。相反,Notch1a细胞内结构域的异位表达会抑制MCC分化。使用DAPT抑制γ-分泌酶表明,在MCC分化模式明显之前,前肾发育早期需要Notch信号传导。令人惊讶的是,我们发现Jagged 2敲低会产生额外的纤毛,并且足以挽救肾纤毛突变体双泡。我们的结果表明,Jagged 2/Notch信号通过涉及抑制rfx2(纤毛生成程序的关键转录调节因子)的遗传途径,调节前肾中多纤毛细胞与转运上皮细胞的数量。