di Porzio U, Rougon G, Novotny E A, Barker J L
Laboratory of Neurophysiology, National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1987 Oct;84(20):7334-8. doi: 10.1073/pnas.84.20.7334.
Dopaminergic neurons represent a rare neurotransmitter phenotype within the mammalian central nervous system. The mesencephalic dopaminergic neurons form the ascending dopaminergic pathways in mammals and are involved in motor and limbic functions. Here we report that about 30% of all developing mouse mesencephalic cells, including virtually all of the dopaminergic phenotype, express surface membrane determinant(s) recognized by a monoclonal antibody to neural specific protein 4 (NSP4). We have been able to isolate and culture neurons from the mesencephalon according to their expression of NSP4, using the anti-NSP4 immunoreaction in conjunction with fluorescence-activated cell sorting. Cultures of NSP4+-sorted cells showed a significant enrichment in three morphologically distinct putative dopaminergic phenotypes when compared to unsorted mesencephalic cultures, whereas the cultures of NSP4--sorted cells were virtually devoid of dopaminergic neurons. This flow cytometric enrichment in dopaminergic neurons should provide the necessary cells for multidisciplinary study of dopaminergic phenotype differentiation.
多巴胺能神经元是哺乳动物中枢神经系统中一种罕见的神经递质表型。中脑多巴胺能神经元在哺乳动物中形成上行多巴胺能通路，并参与运动和边缘系统功能。在此我们报告，所有发育中的小鼠中脑细胞约30%，包括几乎所有多巴胺能表型，表达一种针对神经特异性蛋白4（NSP4）的单克隆抗体所识别的表面膜决定簇。我们能够根据中脑细胞对NSP4的表达，利用抗NSP4免疫反应结合荧光激活细胞分选技术，从中脑分离并培养神经元。与未分选的中脑培养物相比，NSP4+分选细胞的培养物在三种形态上不同的假定多巴胺能表型中显著富集，而NSP4-分选细胞的培养物几乎没有多巴胺能神经元。这种通过流式细胞术对多巴胺能神经元的富集应为多巴胺能表型分化的多学科研究提供所需的细胞。
