Localization of Fos and Jun proteins in rat aortic smooth muscle cells after vascular injury.
Localization of Fos and Jun proteins in rat aortic smooth muscle cells after vascular injury.
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发表时间：
1993-03
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通讯作者：
J. Miano;N. Vlasic;R. R. Tota-R.;Michael B. Stemerman
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作者：
J. Miano;N. Vlasic;R. R. Tota-R.;Michael B. Stemerman
检测基因活性的特异性试剂的问世，为研究体内平滑肌细胞(SMC)增殖提供了重要工具和潜在的新方向。在这篇报道中，我们用Northern印迹法检测了血管损伤后几个FOS和Jun家族成员在主动脉组织中的稳态mRNA水平。此外，对这些基因的蛋白产物进行了免疫细胞化学分析。球囊损伤后15分钟内，c-fos、FosB、c-jun、JunB和Jund的mRNA水平均升高数倍。相反，FOS相关抗原(fra-1)的表达出现延迟。这些即刻早期基因的表达动力学与生长因子刺激的培养细胞的表达动力学相似，提示体内这种SMC基因的激活反映了血源性生长因子向血管壁的渗透或预先形成的生长因子在血管内的释放。球囊损伤后2小时，用抗Fos和Jun蛋白的血清观察到Fos和Jun基因被翻译成免疫反应产物。用放线菌素治疗大鼠后，这种免疫反应消失了。Fos和Jun产物主要分布在大鼠主动脉管腔边缘的SMC核中。这种局部表达可能与血管损伤后SMC DNA合成和迁移的启动有关。此外，血管球囊损伤后SMC中Fos和Jun蛋白的表达可作为多种实验条件下SMC活化的指标。
The availability of specific reagents to measure gene activity has provided important tools and potential new directions for the study of smooth muscle cell (SMC) proliferation in vivo. In this report, we have measured steady-state mRNA levels of several fos and jun family members in aortic tissue by Northern blotting after vascular injury. In addition, protein products of these genes were analyzed by immunocytochemistry. Within 15 minutes of balloon injury, mRNA levels of c-fos, fosB, c-jun, junB, and junD were elevated severalfold. In contrast, fos-related antigen (fra-1) mRNA showed a delayed onset of expression. The expression kinetics of these immediate early genes was similar to those in cultured cells stimulated to undergo proliferation by growth factors, suggesting that such SMC gene activation in vivo reflects permeation of blood-derived growth factors into the vessel wall or intravascular release of preformed growth factors. Translation of fos and jun genes into immunoreactive products was demonstrated 2 hours after balloon injury with antisera to Fos and Jun proteins. Treating rats with cycloheximide abolished this immunoreactivity. The distribution of Fos and Jun products was concentrated in SMC nuclei at the luminal border of the rat aorta. Such focal expression may have consequences for the initiation of SMC DNA synthesis and migration after vascular injury. Furthermore, the expression of Fos and Jun proteins in SMC after vascular balloon injury may be used as an index of SMC activation under a variety of experimental settings.