Kendall G, Ensor E, Brar-Rai A, Winter J, Latchman D S
Department of Molecular Pathology, University College London Medical School, UK.
Brain Res Mol Brain Res. 1994 Aug;25(1-2):73-9. doi: 10.1016/0169-328x(94)90280-1.
We have used primary cultures of adult rat dorsal root ganglia (DRG), enriched in sensory neurons, to investigate the induction of immediate-early genes by NGF and a variety of other growth factors. Using the polymerase chain reaction we have quantitatively amplified specific mRNA transcripts induced by NGF, in the presence and absence of the protein synthesis inhibitor cycloheximide. NGFIA (Egr-1) and NGFIB (nur 77) mRNAs were elevated in level within 60 min of NGF treatment and independently of de novo protein synthesis. This was consistent with the behaviour of immediate-early genes. These kinetics were seen at a range of NGF concentrations. NGFIA and NGFIB mRNAs were also found to be induced in DRG cultures by a variety of other growth factors. Different patterns of induction of NGFIA and NGFIB mRNA observed in DRG cultures suggested that transcript-specific pathways of signal transduction were operating within neurons, dependent upon the particular growth factor stimulus. Comparison of data reported from growth factor treatment of other cell types with data from DRG cultures also revealed patterns of NGFIA and NGFIB mRNA induction specific to DRG neurons.
我们利用富含感觉神经元的成年大鼠背根神经节(DRG)原代培养物,来研究神经生长因子(NGF)及多种其他生长因子对即刻早期基因的诱导作用。我们运用聚合酶链反应,在存在和不存在蛋白质合成抑制剂环己酰亚胺的情况下,对由NGF诱导产生的特定mRNA转录本进行了定量扩增。在NGF处理后60分钟内,NGFIA(Egr-1)和NGFIB(nur 77)的mRNA水平升高,且与从头合成蛋白质无关。这与即刻早期基因的表现一致。在一系列NGF浓度下均观察到了这种动力学现象。在DRG培养物中,多种其他生长因子也能诱导NGFIA和NGFIB的mRNA产生。在DRG培养物中观察到的NGFIA和NGFIB mRNA的不同诱导模式表明,神经元内存在依赖于特定生长因子刺激的转录本特异性信号转导途径。将其他细胞类型经生长因子处理后报告的数据与DRG培养物的数据进行比较,也揭示了DRG神经元特有的NGFIA和NGFIB mRNA诱导模式。