Matsushita Susumu, Ishii Yasuo, Scotting Paul J, Kuroiwa Atsushi, Yasugi Sadao
Department of Biology, School of Medicine, Tokyo Women's Medical University, Shinjuku-ku, Tokyo, Japan.
Dev Dyn. 2002 Jan;223(1):33-47. doi: 10.1002/dvdy.1229.
In this study, we set out to test the ability of endoderm from 1.5-day-old chick embryos (just before digestive tube formation) to develop region-specific characteristics when cultured heterotopically. Various parts of the 1.5-day endoderm were cultured in combination with the flank somatic mesoderm of 3- to 3.5-day chick embryos, and these cultures were analyzed for the expression of several transcription factors and the differentiation of the endoderm. By 1.5 days of normal development, the transcription factors, which are expressed in specific digestive organs, cSox2, CdxA, and cHoxb9/a13 were already expressed in the endodermal cells of the presumptive areas of their later expression domains. When 1.5-day pre-gut endoderm was cultured for 14-15 days, it showed specific differentiation into appropriate organ structures. In general, the more anterior part of the pre-gut endoderm formed the more rostral digestive organ structures while the posterior part became the caudal gut. The differentiation of these regions of endoderm matches their normal fate as recently elucidated (Matsushita [1996a] Rouxs Arch. Dev. Biol. 205:225-231; Matsushita [1999] Dev. Growth Differ. 41:313-319). Expression of cSox2, CdxA, and cHoxb9/a13 in endoderm cultured for 4-5 days is also consistent with their normal fate. Thus, each part of the pre-gut endoderm appears to be already regionally committed to some extent, in accordance with its fate by 1.5 days of development.
在本研究中,我们着手测试1.5日龄鸡胚(就在消化管形成之前)的内胚层在异位培养时形成区域特异性特征的能力。将1.5日龄内胚层的各个部分与3至3.5日龄鸡胚的侧腹体壁中胚层联合培养,并对这些培养物进行几种转录因子表达及内胚层分化的分析。在正常发育到1.5天时,在特定消化器官中表达的转录因子cSox2、CdxA和cHoxb9/a13已经在其后期表达域的预定区域的内胚层细胞中表达。当1.5日龄的前肠内胚层培养14至15天时,它显示出向适当器官结构的特异性分化。一般来说,前肠内胚层越靠前的部分形成越靠头端的消化器官结构,而后部则形成尾肠。内胚层这些区域的分化与其最近阐明的正常命运相符(松下[1996a]《鲁克斯发育生物学杂志》205:225 - 231;松下[1999]《发育、生长与分化》41:313 - 319)。在培养4至5天的内胚层中cSox2、CdxA和cHoxb9/a13的表达也与其正常命运一致。因此,前肠内胚层的每个部分似乎在发育到1.5天时就已经在一定程度上根据其命运进行了区域特化。