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Establishment of cerebellar Purkinje cell lines by somatic cell fusion

通过体细胞融合建立小脑浦肯野细胞系

基本信息

批准号：
03670422
负责人：
NISHIZAWA Masatoyo
金额：
$ 1.28万
依托单位：
Jichi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670422/
关键词：
cerebellum cell fusion neuronal hybridoma degeneration 神経細胞株

项目摘要

In order to obtain clues for elucidating pathogenetic mechanism of cerebellar neurodegenerative disorders, we established murine cerebellar neuronal hybridomas by somatic cell fusion with a neuroblastoma cell line. Several neuronal hybridoma cell lines growing well in vitro and expressing GABA could be identified by immunostaining with anti-GABA antibody. However, we could not detect markers known to be specific for cerebellar Purkinje cells. After culturing these cells under serum-free condition, a part of them also expressed immunoreactivity against tyrosine hydroxylase. This indicates that neuronal properties expressed in these neuronal hybridomas are not always derived from those expressed in parental cerebellar neurons, and that neuronal properties of these hybridomas can change under culture conditions. Since we now realize that neuronal hybridomas are inadequate for studying the expression of neuronal properties, we gave up producing cerebellar neuronal hybridomas from murine models of human cerebellar degenerations, such as pcd, and comparing those with neuronal hybridomas established from normal littermates. Instead, we have made cDNA libraries from these mutant and normal cerebella, and are now trying to identify genes, the expression of which is deferent between them, using differential hybridization method.

为了获得阐明小脑神经退行性疾病发病机制的线索，我们建立了小鼠小脑神经元杂交瘤的体细胞融合与神经母细胞瘤细胞系。用抗GABA抗体免疫组化法鉴定了几株在体外生长良好并表达GABA的神经元杂交瘤细胞系。然而，我们无法检测到已知的小脑浦肯野细胞特异性标志物。在无血清条件下培养这些细胞后，它们中的一部分也表达抗酪氨酸羟化酶的免疫反应性。这表明这些神经元杂交瘤中表达的神经元特性并不总是来源于亲代小脑神经元中表达的神经元特性，并且这些杂交瘤的神经元特性在培养条件下可能会发生变化。由于我们现在意识到神经元杂交瘤不足以研究神经元特性的表达，我们放弃了从人类小脑变性的小鼠模型（如pcd）中产生小脑神经元杂交瘤，并将其与从正常同窝出生的小鼠中建立的神经元杂交瘤进行比较。相反，我们已经从这些突变体和正常小脑中制备了cDNA文库，现在正试图用差异杂交方法鉴定它们之间表达差异的基因。