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Establishment of mouse-immortalized hybrid clones expressing characteristics of differentiated neurons derived from the cerebellar and brain stem regions
Authors:Jun-Ichi Satoh  Ferenc Gallyas  Masumi Endoh  Takashi Yamamura  Tatsuhide Kunishita  Takayoshi Kobayashi  Takeshi Tabira
Abstract:Two clonal immortalized neurons designated CL8c47 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 aceryltransferase 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. © 1992 John Wiley & Sons, Inc.
Keywords:clonal hybrid neurons  differentiation  glutamate  neurofilament proteins  neurite outgrowth
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