Division of Genetic Medicine, Department of Medicine, Vanderbilt University School of Medicine, 2215 Garland Avenue, Nashville, TN 37232-0275, USA.
Hum Mol Genet. 2010 Nov 15;19(22):4353-72. doi: 10.1093/hmg/ddq357. Epub 2010 Aug 25.
Abnormalities in the development of enteric neural crest-derived progenitors (ENPs) that generate the enteric nervous system (ENS) can lead to aganglionosis in a variable portion of the distal gastrointestinal tract. Cumulative evidence suggests that variation of aganglionosis is due to gene interactions that modulate the ability of ENPs to populate the intestine; however, the developmental processes underlying this effect are unknown. We hypothesized that differences in enteric ganglion deficits could be attributable to the effects of genetic background on early developmental processes, including migration, proliferation, or lineage divergence. Developmental processes were investigated in congenic Sox10(Dom) mice, an established Hirschsprung disease (HSCR) model, on distinct inbred backgrounds, C57BL/6J (B6) and C3HeB/FeJ (C3Fe). Immuno-staining on whole-mount fetal gut tissue and dissociated cell suspensions was used to assess migration and proliferation. Flow cytometry utilizing the cell surface markers p75 and HNK-1 was used to isolate live ENPs for analysis of developmental potential. Frequency of ENPs was reduced in Sox10(Dom) embryos relative to wild-type embryos, but was unaffected by genetic background. Both migration and developmental potential of ENPs in Sox10(Dom) embryos were altered by inbred strain background with the most highly significant differences seen for developmental potential between strains and genotypes. In vivo imaging of fetal ENPs and postnatal ganglia demonstrates that altered lineage divergence impacts ganglia in the proximal intestine. Our analysis demonstrates that genetic background alters early ENS development and suggests that abnormalities in lineage diversification can shift the proportions of ENP populations and thus may contribute to ENS deficiencies in vivo.
肠神经嵴祖细胞（ENP）发育异常会导致远端胃肠道的一部分出现无神经节细胞症。越来越多的证据表明，无神经节细胞症的变异是由于基因相互作用，调节了 ENP 定植肠道的能力；然而，这种效应背后的发育过程尚不清楚。我们假设，神经节缺失的差异可能归因于遗传背景对早期发育过程的影响，包括迁移、增殖或谱系分化。在不同的近交系背景下，即 C57BL/6J（B6）和 C3HeB/FeJ（C3Fe）上，对 Sox10(Dom) 小鼠（一种已建立的先天性巨结肠病 [HSCR] 模型）进行了研究。在整个胎肠组织和分离的细胞悬液上进行免疫染色，以评估迁移和增殖。利用细胞表面标志物 p75 和 HNK-1 的流式细胞术用于分离活的 ENP，以分析其发育潜能。Sox10(Dom) 胚胎中的 ENP 频率相对于野生型胚胎减少，但不受遗传背景影响。Sox10(Dom) 胚胎中 ENP 的迁移和发育潜能都受到近交系背景的改变，其中不同品系和基因型之间的发育潜能差异最为显著。对胎生 ENP 和出生后神经节的体内成像表明，谱系分化的改变会影响近端肠道的神经节。我们的分析表明遗传背景改变了早期 ENS 发育，并表明谱系多样化的异常可能会改变 ENP 群体的比例，从而可能导致体内 ENS 缺陷。
