Shinar Y, McMorris F A
Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
J Neurosci Res. 1995 Nov 1;42(4):516-27. doi: 10.1002/jnr.490420410.
Insulin-like growth factors IGF-I and IGF-II are potent inducers of oligodendrocyte development. Because IGF-I is produced, in some cases, by the same cells that respond to it (autocrine/paracrine action), we examined the possibility that IGF-I is expressed by developing oligodendroglial cells. We employed a sensitive method, reverse transcriptase-polymerase chain reaction (RT-PCR), to detect IGF-I mRNA in purified populations of oligodendroglial cells isolated from rat brain during the period of oligodendrocyte development. Cells were purified by fluorescence activated cell sorting (FACS), using antibodies to the cell surface antigenic markers O4 and galactocerebroside (GC). RNA was isolated from the sorted cells, reverse-transcribed, and PCR-amplified, using a strategy that recognizes IGF-I mRNA but not DNA. The amplified band was identified as IGF-I by size, hybridization to an IGF-I-specific antisense probe, and restriction analysis. IGF-I mRNA was detected in O4-positive/GC-negative oligodendrocyte precursors and, more weakly, in GC-positive oligodendrocytes. IGF-I mRNA could be detected reproducibly in RNA extracted from 100-cell samples of O4-positive cells, making it unlikely that the mRNA was derived from contaminants in the FACS-sorted cell populations. We conclude that IGF-I is expressed by developing oligodendroglia. Autocrine expression of IGF-I by developing oligodendroglial cells suggests that oligodendrocyte development is, in part, autoregulatory.
胰岛素样生长因子IGF-I和IGF-II是少突胶质细胞发育的有效诱导剂。由于在某些情况下,IGF-I是由对其产生反应的相同细胞产生的(自分泌/旁分泌作用),我们研究了发育中的少突胶质细胞是否表达IGF-I。我们采用了一种灵敏的方法,即逆转录聚合酶链反应(RT-PCR),来检测在少突胶质细胞发育期间从大鼠脑中分离出的纯化少突胶质细胞群体中的IGF-I mRNA。使用针对细胞表面抗原标志物O4和半乳糖脑苷脂(GC)的抗体,通过荧光激活细胞分选(FACS)对细胞进行纯化。从分选的细胞中分离RNA,进行逆转录,并使用一种识别IGF-I mRNA而不识别DNA的策略进行PCR扩增。通过大小、与IGF-I特异性反义探针杂交以及限制性分析,将扩增带鉴定为IGF-I。在O4阳性/GC阴性少突胶质细胞前体中检测到IGF-I mRNA,在GC阳性少突胶质细胞中检测到的信号较弱。在从100个O4阳性细胞样本中提取的RNA中可重复检测到IGF-I mRNA,这使得该mRNA不太可能来自FACS分选细胞群体中的污染物。我们得出结论,发育中的少突胶质细胞表达IGF-I。发育中的少突胶质细胞自分泌IGF-I表明少突胶质细胞发育在一定程度上是自我调节的。