Lack of organ specific commitment of vagal neural crest cell derivatives as shown by back-transplantation of GFP chicken tissues
Lack of organ specific commitment of vagal neural crest cell derivatives as shown by back-transplantation of GFP chicken tissues
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DOI：
10.1387/ijdb.113438lf
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发表时间：
2012-01-01
影响因子：
0.7
通讯作者：
Burns, Alan J.
中科院分区：
文献类型：
--
作者：
Freem, Lucy J.;Delalande, Jean Marie;Burns, Alan J.
神经嵴细胞（NCC）是一种多能祖细胞，在胚胎发育过程中广泛迁移，并产生多种细胞类型。迷走神经细胞从后脑迁移到前肠，并从那里沿着胃肠道形成肠神经系统（ENS），即肠道的内在神经支配，并进入发育中的肺芽形成肺的内在神经支配。本研究的目的是确定已经在肠道或肺部定植的迷走神经细胞癌的发育潜力。我们利用普遍表达绿色荧光蛋白（GFP）的转基因鸡胚胎，采用种内嫁接技术永久标记迷走神经细胞癌并绘制其命运图。结合将含有gfp阳性NCC的肠道和肺段反移植到第二个受体胚胎的迷走神经区，使用免疫组织化学染色来确定肠道或肺NCC是否能够在这两个器官重新定植，或者它们的命运是否受到限制。鸡(GFP)-鸡种内移植在鸡胚肠和肺内有效标记NCC。当含有gfp阳性NCC的胚胎日(E)5.5脐带前中肠片段被移植到E1.5宿主胚胎的迷走神经区时，gfp阳性NCC迁移到肠道和肺部，并在典型位置分化为神经元。然而，gfp阳性的肺NCC在回移植时不会迁移。我们的研究表明，肠道NCC不仅局限于定植这个器官，因为在背部移植时，gfp阳性的肠道NCC在肠道和肺部都定植。
Neural crest cells (NCC) are multipotent progenitors that migrate extensively throughout the developing embryo and generate a diverse range of cell types. Vagal NCC migrate from the hindbrain into the foregut and from there along the gastrointestinal tract to form the enteric nervous system (ENS), the intrinsic innervation of the gut, and into the developing lung buds to form the intrinsic innervation of the lungs. The aim of this study was to determine the developmental potential of vagal NCC that had already colonised the gut or the lungs. We used transgenic chicken embryos that ubiquitously express green fluorescent protein (GFP) to permanently mark and fate-map vagal NCC using intraspecies grafting. This was combined with back-transplantation of gut and lung segments, containing GFP-positive NCC, into the vagal region of a second recipient embryo to determine, using immunohistochemical staining, whether gut or lung NCC are competent of re-colonising both these organs, or whether their fate is restricted. Chick(GFP)-chick intraspecies grafting efficiently labelled NCC within the gut and lung of chick embryos. When segments of embryonic day (E)5.5 pre-umbilical midgut containing GFP-positive NCC were back-transplanted into the vagal region of E1.5 host embryos, the GFP-positive NCC remigrated to colonise both the gut and lungs and differentiated into neurons in stereotypical locations. However, GFP-positive lung NCC did not remigrate when back-transplanted. Our studies suggest that gut NCC are not restricted to colonising only this organ, since upon back-transplantation GFP-positive gut NCC colonised both the gut and the lung.