Centre de Ressources Biologiques Xénope UMS U3387, CNRS, and Rennes 1 University, Rennes, France.
PLoS One. 2013 Jun 14;8(6):e66487. doi: 10.1371/journal.pone.0066487. Print 2013.
In contrast to mammals that have limited proliferation and neurogenesis capacities, the Xenopus frog exhibit a great potential regarding proliferation and production of new cells in the adult brain. This ability makes Xenopus a useful model for understanding the molecular programs required for adult neurogenesis. Transcriptional factors that control adult neurogenesis in vertebrate species undergoing widespread neurogenesis are unknown. NeuroD1 is a member of the family of proneural genes, which function during embryonic neurogenesis as a potent neuronal differentiation factor. Here, we study in detail the expression of NeuroD1 gene in the juvenile and adult Xenopus brains by in situ hybridization combined with immunodetections for proliferation markers (PCNA, BrdU) or in situ hybridizations for cell type markers (Vimentin, Sox2). We found NeuroD1 gene activity in many brain regions, including olfactory bulbs, pallial regions of cerebral hemispheres, preoptic area, habenula, hypothalamus, cerebellum and medulla oblongata. We also demonstrated by double staining NeuroD1/BrdU experiments, after long post-BrdU administration survival times, that NeuroD1 gene activity was turned on in new born neurons during post-metamorphic neurogenesis. Importantly, we provided evidence that NeuroD1-expressing cells at this brain developmental stage were post-mitotic (PCNA-) cells and not radial glial (Vimentin+) or progenitors (Sox2+) cells.
与增殖和神经发生能力有限的哺乳动物相比,非洲爪蟾在成年大脑中的增殖和新细胞产生方面表现出巨大的潜力。这种能力使非洲爪蟾成为理解成年神经发生所需的分子程序的有用模型。在经历广泛神经发生的脊椎动物物种中,控制成年神经发生的转录因子尚不清楚。NeuroD1 是神经前基因家族的成员,在胚胎神经发生过程中作为一种有效的神经元分化因子发挥作用。在这里,我们通过原位杂交结合增殖标志物(PCNA、BrdU)的免疫检测或细胞类型标志物(Vimentin、Sox2)的原位杂交,详细研究了 NeuroD1 基因在幼年和成年非洲爪蟾大脑中的表达。我们发现 NeuroD1 基因在许多脑区都有活性,包括嗅球、大脑半球的皮层区、视前区、缰核、下丘脑、小脑和延髓。我们还通过双重染色 NeuroD1/BrdU 实验证明,在 BrdU 给药后长时间存活后,NeuroD1 基因活性在变态后神经发生期间被开启于新生神经元中。重要的是,我们提供的证据表明,在这个大脑发育阶段表达 NeuroD1 的细胞是有丝分裂后(PCNA-)细胞,而不是放射状胶质(Vimentin+)或祖细胞(Sox2+)细胞。