Lack of (cid:3) 1 integrins in enteric neural crest cells leads to a Hirschsprung-like phenotype
Lack of (cid:3) 1 integrins in enteric neural crest cells leads to a Hirschsprung-like phenotype
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肠神经嵴细胞中缺乏 (cid:3) 1 整合素会导致先天性巨结肠表型
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通讯作者：
P. Talbot
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作者：
G. Melkonian;J. L. Wang;J. Chung;N. Munoz;P. Talbot
作者：Pietri 等人。 （Pietri 等人，2004）对于整个组织和石蜡切片，以及 Delannet 等人。 (Delannet et al., 1994) 用于冰冻切片。我们使用的一抗列于补充材料的表 S1 中。 (cid:3)-半乳糖苷酶 ((cid:3)-gal) 活性通过对整个组织进行 X-Gal 染色来检测，如 Dufour 等人所述。 （杜福尔等人，1994）。使用(cid:3)-半乳糖苷酶测定系统(Promega)测量蛋白质提取物的(cid:3)-gal活性。肠神经系统主要由迷走神经和骶神经嵴细胞产生，这些细胞在小鼠发育的 9.5 至 14 天之间定植于肠道。使用 Cre-LoxP 系统，当神经嵴细胞从神经管中出现时，我们去除了神经嵴细胞中的 (cid:3) 1 整合素。 (cid:3) 1-null 肠神经嵴细胞无法完全定植于肠道，导致降结肠神经节缺失，类似于人类先天性巨结肠症。此外，(cid:3) 1-null肠神经嵴细胞在肠壁中形成异常聚集体，导致神经节网络组织的严重改变。肠道外植体的器官型培养显示，(cid:3) 1-null 肠神经嵴细胞表现出细胞外基质粘附受损和细胞间粘附特性增强。它们在胶原凝胶和肠道组织环境中表现出迁移缺陷。我们还提供证据表明 (cid:3) 1 整合素是小肠绒毛神经支配所必需的。我们的研究结果强调了 (cid:3)1 整合素在肠神经系统发育的各个步骤中发挥的关键作用。
by Pietri et al. (Pietri et al., 2004) for whole tissues and paraffin sections, and by Delannet et al. (Delannet et al., 1994) for frozen sections. The primary antibodies we used are listed in Table S1 in the supplementary material. The (cid:3) -galactosidase ( (cid:3) -gal) activity was detected by X-Gal staining on whole tissues as described in Dufour et al. (Dufour et al., 1994). The (cid:3) -gal activity of the protein extracts was measured using the (cid:3) -galactosidase Enzyme Assay System (Promega). The enteric nervous system arises mainly from vagal and sacral neural crest cells that colonise the gut between 9.5 and 14 days of development in mice. Using the Cre-LoxP system, we removed (cid:3) 1 integrins in the neural crest cells when they emerge from the neural tube. (cid:3) 1-null enteric neural crest cells fail to colonise the gut completely, leading to an aganglionosis of the descending colon, which resembles the human Hirschsprung’s disease. Moreover, (cid:3) 1-null enteric neural crest cells form abnormal aggregates in the gut wall, leading to a severe alteration of the ganglia network organisation. Organotypic cultures of gut explants reveal that (cid:3) 1-null enteric neural crest cells show impaired adhesion on extracellular matrix and enhanced intercellular adhesion properties. They display migration defects in collagen gels and gut tissue environments. We also provide evidence that (cid:3) 1 integrins are required for the villi innervation in the small intestine. Our findings highlight the crucial roles played by (cid:3) 1 integrins at various steps of enteric nervous system development.