Charrier C, Coronas V, Fombonne J, Roger M, Jean A, Krantic S, Moyse E
Physiologie Neurovégétative, UMR CNRS (6153), INRA (1147), Université Aix-Marseille III Paul Cézanne, France.
Neuroscience. 2006;138(1):5-16. doi: 10.1016/j.neuroscience.2005.10.046. Epub 2005 Dec 7.
The dorsal vagal complex, located in the brainstem, is the major integrative center of the autonomic nervous system. By combining in vivo bromodeoxyuridine incorporation and phenotypic immunolabeling, we have previously reported that neurogenesis occurs in the adult rat dorsal vagal complex [Bauer S, Hay M, Amilhon B, Jean A, Moyse E (2005) In vivo neurogenesis in the dorsal vagal complex of the adult rat brainstem. Neuroscience 130:75-90.]. In the present study we asked whether adult dorsal vagal complex contains proliferative and/or neural stem cells. Using Ki-67 immunolabeling and cyclin D1 Western blot, we showed intrinsic cell proliferation in the dorsal vagal complex and its stimulation by vagotomy. Detailed time-course analysis revealed that vagotomy-induced proliferation in the dorsal vagal complex peaked three days after lesion. In order to directly assess the presence of intrinsic stem cells, primary cell cultures from adult rat dorsal vagal complex were performed in the presence of epidermal growth factor and basic fibroblast growth factor (neurosphere assay). A discrete subpopulation of dorsal vagal complex cells proliferated as neurospheres, self-renewed when passaged, and differentiated into neurons, astrocytes and oligodendrocytes. Proliferation and neuron-differentiating potentials of dorsal vagal complex neurospheres were both lower than those of subventricular zone neurospheres from the same rats. The relationship between in vitro neurosphere-forming cells of dorsal vagal complex and in vivo dorsal vagal complex neurogenesis is discussed and remains to be directly addressed. The present data demonstrate the occurrence of neural stem cells in the dorsal vagal complex of adult rat brain.
位于脑干的迷走神经背核复合体是自主神经系统的主要整合中心。通过结合体内溴脱氧尿苷掺入和表型免疫标记，我们之前报道过成年大鼠迷走神经背核复合体内发生神经发生[鲍尔 S，海 M，阿米隆 B，让 A，莫伊斯 E（2005年）成年大鼠脑干迷走神经背核复合体内的体内神经发生。神经科学130：75 - 90]。在本研究中，我们探究成年迷走神经背核复合体是否包含增殖性和/或神经干细胞。使用Ki - 67免疫标记和细胞周期蛋白D1免疫印迹法，我们显示了迷走神经背核复合体内的固有细胞增殖以及迷走神经切断术对其的刺激作用。详细的时间进程分析表明，迷走神经切断术诱导的迷走神经背核复合体增殖在损伤后三天达到峰值。为了直接评估固有干细胞的存在情况，在表皮生长因子和碱性成纤维细胞生长因子存在的条件下进行了成年大鼠迷走神经背核复合体的原代细胞培养（神经球测定）。迷走神经背核复合体细胞的一个离散亚群增殖形成神经球，传代时自我更新，并分化为神经元、星形胶质细胞和少突胶质细胞。迷走神经背核复合体神经球的增殖和神经元分化潜能均低于同一只大鼠脑室下区神经球。讨论了迷走神经背核复合体体外形成神经球的细胞与体内迷走神经背核复合体神经发生之间的关系，该关系仍有待直接研究。目前的数据证明成年大鼠脑的迷走神经背核复合体内存在神经干细胞。
