Pituello F, Kan P, Geffard M, Duprat A M
URA-CNRS 675 l'INSERM, Université Paul Sabatier, Toulouse, France.
Int J Dev Biol. 1989 Dec;33(4):445-53.
At the late gastrula-early neurula stage some embryonic neuroblasts from neural plate and neural fold present apparently as a consequence of neural induction, the capability to develop in vitro into different neuronal subpopulations (cholinergic, dopaminergic, noradrenergic, somatostatinergic and some other peptidergic subpopulations without ongoing influences from the chordamesoderm (Duprat et al., 1987). Using the same in vitro model system, the aim of the present work was to delineate the abilities of these neuroblasts to develop GABAergic traits. The initial appearance and development of GABAergic phenotype has been quantitated by assaying the activity of glutamic acid decarboxylase (GAD). GAD activity was undetectable at the early gastrula stage (stage 8a) and was slightly measurable at the early neurula stage (stage 14- onset of the culture). It increased subsequently over the next 14 days in vitro. The temporal pattern of appearance and development of GAD activity in culture was in agreement with that observed in vivo. Immunocytochemical studies showed that GABA-like immunoreactivity was expressed in vitro in a subpopulation of neurons. Thus the developmental program for GAD expression and GABA phenotype maturation is acquired at least in some neuronal precursors. These data together with previously reported results on the expression of cholinergic, catecholaminergic and peptidergic phenotypes demonstrate that different neuronal subpopulations emerge near the end of gastrulation i.e. immediately after neural induction. The embryonic origin of this neuroblast heterogeneity remains to be determined.
在原肠胚晚期至神经胚早期阶段,由于神经诱导作用,神经板和神经褶中的一些胚胎神经母细胞明显出现,它们具有在体外发育成不同神经元亚群(胆碱能、多巴胺能、去甲肾上腺素能、生长抑素能以及其他一些肽能亚群)的能力,且无需脊索中胚层的持续影响(Duprat等人,1987年)。利用相同的体外模型系统,本研究的目的是描绘这些神经母细胞发育出γ-氨基丁酸能特性的能力。通过检测谷氨酸脱羧酶(GAD)的活性,对γ-氨基丁酸能表型的最初出现和发育进行了定量分析。在原肠胚早期阶段(8a期)未检测到GAD活性,在神经胚早期阶段(14期 - 培养开始时)可略微检测到。随后在接下来的14天体外培养中其活性增加。培养物中GAD活性出现和发育的时间模式与体内观察到的一致。免疫细胞化学研究表明,γ-氨基丁酸样免疫反应性在体外的一部分神经元中表达。因此,至少在一些神经元前体中获得了GAD表达和γ-氨基丁酸表型成熟的发育程序。这些数据与先前报道的关于胆碱能、儿茶酚胺能和肽能表型表达的结果一起表明,不同的神经元亚群在原肠胚形成末期即神经诱导后立即出现。这种神经母细胞异质性的胚胎起源仍有待确定。