Freem Lucy J, Delalande Jean Marie, Campbell Alison M, Thapar Nikhil, Burns Alan J
Neural Development and Gastroenterology Units, UCL Institute of Child Health, London, UK.
Int J Dev Biol. 2012;56(4):245-54. doi: 10.1387/ijdb.113438lf.
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.
神经嵴细胞(NCC)是多能祖细胞,在整个发育中的胚胎中广泛迁移,并产生多种细胞类型。迷走神经嵴细胞从后脑迁移到前肠,然后沿着胃肠道迁移,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定殖于肠道或肺部的迷走神经嵴细胞的发育潜能。我们使用了普遍表达绿色荧光蛋白(GFP)的转基因鸡胚胎,通过种内移植来永久标记和追踪迷走神经嵴细胞的命运。这与含有GFP阳性神经嵴细胞的肠道和肺段的回植相结合,将其移植到第二个受体胚胎的迷走神经区域,通过免疫组织化学染色来确定肠道或肺神经嵴细胞是否有能力重新定殖这两个器官,或者它们的命运是否受到限制。鸡(GFP)-鸡种内移植有效地标记了鸡胚胎肠道和肺内的神经嵴细胞。当将含有GFP阳性神经嵴细胞的胚胎第(E)5.5天脐前中肠段回植到E1.5宿主胚胎的迷走神经区域时,GFP阳性神经嵴细胞重新迁移,定殖于肠道和肺部,并在典型位置分化为神经元。然而,GFP阳性肺神经嵴细胞回植时不会重新迁移。我们的研究表明,肠道神经嵴细胞并不局限于仅定殖于该器官,因为回植后GFP阳性肠道神经嵴细胞定殖于肠道和肺部。