Satoh J, Gallyas F, Endoh M, Yamamura T, Kunishita T, Kobayashi T, Tabira T
Division of Demyelinating Diseases and Aging, National Institute of Neuroscience, Tokyo, Japan.
J Neurobiol. 1992 Sep;23(7):905-19. doi: 10.1002/neu.480230711.
Two clonal immortalized neurons designated CL8c4.7 and CL8a5.2 were established by somatic cell fusion between a hypoxanthine phosphoribosyltransferase-(HPRT-) deficient neuroblastoma N18TG2 and newborn mouse cerebellar/brain stem neurons. In the serum-containing medium without extra differentiating agents, both clones exhibited a morphology of differentiated neurons. They contained high levels of glutamate but no gamma-aminobutyric acid (GABA). The CL8a5.2 clone synthesized choline acetyltransferase and serotonin. In immunocytochemical studies, both clones expressed 200 kD neurofilament protein, neuron-specific enolase, microtubule-associated protein 2 (MAP2), tau protein, neuronal cell adhesion molecule (N-CAM), HNK-1, Thy-1.2, saxitoxin-binding sodium channel protein, and glutamate. Synaptophysin immunoreactivity was identified in the neuritic terminals of CL8c4.7 cells. Most of these antigens were barely detectable on N18TG2 cells. Electrophysiologically, both clones generated action potentials in response to electrical stimuli. The hybrid clones that express characteristics of differentiated neurons derived from the cerebellar and brain stem regions might be invaluable for the study of the molecular basis of neuronal differentiation and degeneration in these regions.
通过次黄嘌呤磷酸核糖基转移酶（HPRT）缺陷的神经母细胞瘤N18TG2与新生小鼠小脑/脑干神经元之间的体细胞融合，建立了两个克隆永生神经元，分别命名为CL8c4.7和CL8a5.2。在不含额外分化剂的含血清培养基中，两个克隆均呈现出分化神经元的形态。它们含有高水平的谷氨酸，但没有γ-氨基丁酸（GABA）。CL8a5.2克隆合成胆碱乙酰转移酶和5-羟色胺。在免疫细胞化学研究中，两个克隆均表达200 kD神经丝蛋白、神经元特异性烯醇化酶、微管相关蛋白2（MAP2）、tau蛋白、神经元细胞黏附分子（N-CAM）、HNK-1、Thy-1.2、石房蛤毒素结合钠通道蛋白和谷氨酸。在CL8c4.7细胞的神经末梢中鉴定出突触素免疫反应性。在N18TG2细胞上几乎检测不到这些抗原中的大多数。在电生理方面，两个克隆均能对电刺激产生动作电位。表达源自小脑和脑干区域的分化神经元特征的杂交克隆，对于研究这些区域神经元分化和退化的分子基础可能具有重要价值。
