A novel mouse model reveals that polycystin-1 deficiency in ependyma and choroid plexus results in dysfunctional cilia and hydrocephalus.
A novel mouse model reveals that polycystin-1 deficiency in ependyma and choroid plexus results in dysfunctional cilia and hydrocephalus.
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DOI：
10.1371/journal.pone.0007137
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发表时间：
2009-09-23
期刊：
影响因子：
3.7
通讯作者：
Boletta A
中科院分区：
文献类型：
--
作者：
Wodarczyk C;Rowe I;Chiaravalli M;Pema M;Qian F;Boletta A
多囊蛋白-1 (PC-1) 是 PKD1 基因的产物，在大多数常染色体显性多囊肾病 (ADPKD) 病例中发生突变，是一种非常大 (∼520 kDa) 的质膜受体，位于多个亚细胞区室中，包括细胞-细胞/基质连接以及纤毛。虽然异源过表达系统已经允许鉴定该受体的几种潜在生物学作用，但其精确功能仍然很大程度上难以捉摸。由于 PC-1 的复杂性和低表达水平，体内研究 PC-1 一直是一项具有挑战性的任务。为了克服这些限制并促进内源性 PC-1 的研究，我们通过同源重组将 HA 或 Myc 标签序列插入 Pkd1 基因座。在这里，我们表明我们的方法成功地生成了功能齐全且易于检测的内源性 PC-1。 PC-1 体内分布的表征表明，它在大多数组织中普遍表达并受到发育调节。此外，我们的新工具使我们能够研究 PC-1 在大脑中的作用，该蛋白质在大脑中大量表达。 PC-1 的亚细胞定位揭示了纤毛室管膜和脉络丛细胞中强烈且特异性的染色。与这种分布一致，我们在普遍存在的敲除胚胎和大脑中 Pkd1 基因条件性失活的新生小鼠中观察到脑积水的形成。这些小鼠的脉络丛和室管膜纤毛形态均正常，表明 PC-1 在该区室的纤毛功能或信号传导中发挥作用，而不是在纤毛发生中发挥作用。我们认为 PC-1 在大脑纤毛中的作用可能是预防脑积水，这是该受体以前未被认识到的作用，并且可能对其他遗传或散发性疾病具有重要意义。
Polycystin-1 (PC-1), the product of the PKD1 gene, mutated in the majority of cases of Autosomal Dominant Polycystic Kidney Disease (ADPKD), is a very large (∼520 kDa) plasma membrane receptor localized in several subcellular compartments including cell-cell/matrix junctions as well as cilia. While heterologous over-expression systems have allowed identification of several of the potential biological roles of this receptor, its precise function remains largely elusive. Studying PC-1 in vivo has been a challenging task due to its complexity and low expression levels. To overcome these limitations and facilitate the study of endogenous PC-1, we have inserted HA- or Myc-tag sequences into the Pkd1 locus by homologous recombination. Here, we show that our approach was successful in generating a fully functional and easily detectable endogenous PC-1. Characterization of PC-1 distribution in vivo showed that it is expressed ubiquitously and is developmentally-regulated in most tissues. Furthermore, our novel tool allowed us to investigate the role of PC-1 in brain, where the protein is abundantly expressed. Subcellular localization of PC-1 revealed strong and specific staining in ciliated ependymal and choroid plexus cells. Consistent with this distribution, we observed hydrocephalus formation both in the ubiquitous knock-out embryos and in newborn mice with conditional inactivation of the Pkd1 gene in the brain. Both choroid plexus and ependymal cilia were morphologically normal in these mice, suggesting a role for PC-1 in ciliary function or signalling in this compartment, rather than in ciliogenesis. We propose that the role of PC-1 in the brain cilia might be to prevent hydrocephalus, a previously unrecognized role for this receptor and one that might have important implications for other genetic or sporadic diseases.
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影响因子：
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