Jordan C, Friedrich V, Dubois-Dalcq M
Laboratory of Molecular Genetics, National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, Maryland 20892.
J Neurosci. 1989 Jan;9(1):248-57. doi: 10.1523/JNEUROSCI.09-01-00248.1989.
We analyzed the location and abundance of transcripts for the 4 CNS myelin protein genes, myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG), and 2',3'-cyclic nucleotide phosphohydrolase (CNP), in the mouse cervical spinal cord from the time of rapid myelination until adulthood (8-45 d). In the white matter, maximal levels of transcripts were found for each of the myelin genes at the peak of myelination (8 d). Total MBP and PLP mRNAs stayed high until 20 d and showed a minor decrease thereafter. In contrast, MAG and the MBP exon 2 containing transcripts (coding for the 21.5 and 17 kDa MBP isoforms) decreased sharply between 8 and 20 d, suggesting that high levels of these transcripts are needed primarily during the initiation of myelination. CNP transcripts were less abundant, maintained high expression until 20 d, and then decreased sharply. PLP, MAG, and CNP transcripts were clustered in the oligodendrocyte cell body, while MBP mRNAs were scattered throughout the cell body and processes. In contrast to the white matter, all these myelin specific transcripts in the gray matter showed a marked increase from 8 to 20 d, as did the number of oligodendrocytes identified by CNP immunostaining. MAG transcripts were found in white matter and in satellite and other oligodendrocytes of the gray matter but not in neurons identified by their expression of neurofilament transcripts. The results of our quantitative in situ hybridization study are in good agreement with those of previous molecular studies and provide new information on the cellular and topographic distribution of myelin-specific mRNAs during myelination.
我们分析了4种中枢神经系统髓鞘蛋白基因,即髓鞘碱性蛋白(MBP)、蛋白脂蛋白(PLP)、髓鞘相关糖蛋白(MAG)和2',3'-环核苷酸磷酸二酯酶(CNP)的转录本在小鼠颈脊髓从快速髓鞘形成期直至成年期(8 - 45日龄)的位置和丰度。在白质中,每种髓鞘基因的转录本在髓鞘形成高峰期(8日龄)达到最高水平。总MBP和PLP mRNA在20日龄前一直保持较高水平,此后略有下降。相比之下,MAG和含有MBP外显子2的转录本(编码21.5 kDa和17 kDa的MBP异构体)在8至20日龄之间急剧下降,这表明这些转录本的高水平主要在髓鞘形成起始阶段需要。CNP转录本丰度较低,在20日龄前保持高表达,然后急剧下降。PLP、MAG和CNP转录本聚集在少突胶质细胞胞体中,而MBP mRNA则散布在整个胞体和突起中。与白质不同,灰质中所有这些髓鞘特异性转录本从8日龄到20日龄均显著增加,通过CNP免疫染色鉴定的少突胶质细胞数量也如此。在白质以及灰质的卫星少突胶质细胞和其他少突胶质细胞中发现了MAG转录本,但在通过神经丝转录本表达鉴定的神经元中未发现。我们定量原位杂交研究的结果与先前的分子研究结果高度一致,并提供了关于髓鞘形成过程中髓鞘特异性mRNA的细胞和拓扑分布的新信息。