Feutz A C, Bellomi I, Allinquant B, Schladenhaufen Y, Ghandour M S
Laboratoire de Neurobiologie Ontogénique (UPR 417-CNRS), Centre de Neurochimie, Strasbourg, France.
J Neurocytol. 1995 Nov;24(11):865-77. doi: 10.1007/BF01179985.
This study characterizes jimpy oligodendrocyte-enriched secondary cultures isolated from 10-12 days in vitro primary glial cell cultures derived from 1-2-day-old jimpy mouse brains. Proliferation of defective oligodendrocytes was carefully investigated with regard to the expression of myelin basic protein and proteolipid protein and their respective mRNAs. Less than 5% of contaminating astrocytes (GFAP+ cells) were usually present. The identity of jimpy oligodendrocytes was confirmed using an antibody directed against a peptide from the wild type proteolipid protein C-terminal sequence for immunocytochemistry and an oligonucleotide complementary to mRNA derived from exon 5 of the proteolipid protein gene for in situ hybridization. Both the antibody and the probe recognize only normal oligondedrocytes while jimpy oligodendrocytes always remain unstained. Proteolipid protein in normal and jimpy oligodendrocytes was detected with antibody recognizing normal and mutated forms. Between 80 and 95% of the cells in normal and jimpy cultures at 2 and 4 days in vitro in secondary cultures express myelin basic protein and proteolipid protein and their respective mRNAs. The percentage of oligodendrocytes (PLP+ or MBP+) in S phase of the cell cycle was 7-10% for both normal and jimpy oligodendrocytes. This contrasts with the in vivo situation where the proliferation rate of oligodendrocytes in jimpy brains is higher than in normal brains. In addition, jimpy oligodendrocytes remain unresponsive to basic fibroblast growth factor treatment while a similar treatment stimulates the proliferation of normal oligodendrocytes.
本研究对从1 - 2日龄jimpy小鼠脑来源的体外培养10 - 12天的原代神经胶质细胞培养物中分离得到的富含jimpy少突胶质细胞的传代培养物进行了表征。针对髓鞘碱性蛋白和蛋白脂质蛋白及其各自mRNA的表达，仔细研究了有缺陷的少突胶质细胞的增殖情况。通常存在的污染星形胶质细胞（GFAP +细胞）不到5%。使用针对野生型蛋白脂质蛋白C末端序列的肽的抗体进行免疫细胞化学，以及使用与蛋白脂质蛋白基因外显子5衍生的mRNA互补的寡核苷酸进行原位杂交，证实了jimpy少突胶质细胞的身份。该抗体和探针仅识别正常少突胶质细胞，而jimpy少突胶质细胞始终不着色。用识别正常和突变形式的抗体检测正常和jimpy少突胶质细胞中的蛋白脂质蛋白。在传代培养的第2天和第4天，正常和jimpy培养物中80%至95%的细胞表达髓鞘碱性蛋白和蛋白脂质蛋白及其各自的mRNA。正常和jimpy少突胶质细胞在细胞周期S期的少突胶质细胞百分比（PLP +或MBP +）均为7 - 10%。这与体内情况形成对比，在jimpy脑内少突胶质细胞的增殖率高于正常脑。此外，jimpy少突胶质细胞对碱性成纤维细胞生长因子处理无反应，而类似处理可刺激正常少突胶质细胞的增殖。
