Du Pasquier D, Chesneau A, Ymlahi-Ouazzani Q, Boistel R, Pollet N, Ballagny C, Sachs L M, Demeneix B, Mazabraud A
Laboratoire de Transgenèse et Génétique des Amphibiens, Université Paris-Sud, Orsay, France.
Genesis. 2007 Jan;45(1):1-10. doi: 10.1002/dvg.20252.
Xenopus is a well proven model for a wide variety of developmental studies, including cell lineage. Cell lineage in Xenopus has largely been addressed by injection of tracer molecules or by micro-dissection elimination of blastomeres. Here we describe a genetic method for cell ablation based on the use of tBid, a direct activator of the mitochondrial apoptotic pathway. In mammalian cells, cross-talk between the main apoptotic pathways (the mitochondrial and the death domain protein pathways) involve the pro-death protein BID, the active form of which, tBID, results from protease truncation and translocation to mitochondria. In transgenic Xenopus, restricting tBID expression to the lens-forming cells enables the specific ablation of the lens without affecting the development of other eye structures. Thus, overexpression of tBid can be used in vivo as a tool to eliminate a defined cell population by apoptosis in a developing organism and to evaluate the degree of autonomy or the inductive effects of a specific tissue during embryonic development.
非洲爪蟾是用于包括细胞谱系在内的多种发育研究的成熟模型。非洲爪蟾的细胞谱系很大程度上是通过注射示踪分子或通过显微解剖去除卵裂球来研究的。在此，我们描述一种基于使用tBid（线粒体凋亡途径的直接激活剂）进行细胞消融的遗传方法。在哺乳动物细胞中，主要凋亡途径（线粒体和死亡结构域蛋白途径）之间的相互作用涉及促凋亡蛋白BID，其活性形式tBID是蛋白酶切割并转运至线粒体的结果。在转基因非洲爪蟾中，将tBid的表达限制在晶状体形成细胞中可实现晶状体的特异性消融，而不影响其他眼结构的发育。因此，tBid的过表达可在体内用作一种工具，通过凋亡消除发育生物体中特定的细胞群体，并评估胚胎发育过程中特定组织的自主程度或诱导作用。
