Meiners S, Marone M, Rittenhouse J L, Geller H M
Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854.
Dev Biol. 1993 Dec;160(2):480-93. doi: 10.1006/dbio.1993.1322.
Extracellular matrix (ECM) molecules have been implicated in the regulation of neuronal adhesion and neurite outgrowth both during development and after injury. It has been demonstrated in our laboratory that astrocytes are heterogeneous in expression of the ECM molecule tenascin. High-tenascin astrocytes have a reduced ability to support neurite outgrowth. In addition, astrocytes treated with exogenous basic fibroblast growth factor (bFGF) supported reduced neuronal growth and adhesion. In the current study, the hypothesis was tested that bFGF could increase the expression of tenascin by these cells. Basic FGF was added to cultures of rat cerebral cortical astrocytes at concentrations of up to 30 ng/ml, concentrations shown to have a significant effect on neuronal adhesion. Tenascin levels were evaluated by Western blot analysis of both cell extracts and conditioned media and also by immunocytochemistry techniques. Tenascin levels began to increase after 24-48 hr and continued to increase throughout 8 days in culture. The increase in tenascin was concentration-dependent, with the largest increase seen at 5 ng/ml bFGF. Tenascin production was increased approximately 5.5-fold in serum-containing medium but only about 2-fold in serum-free medium. When heparin (10 micrograms/ml) was included along with bFGF in serum-free medium, tenascin production was further enhanced. The bFGF treatment was discontinued after 8 days, and the cells were maintained for an additional 8 days in culture. Tenascin levels returned to control values, demonstrating that the bFGF effect is transient. It is our hypothesis that the action of bFGF during injury may evoke the induction of tenascin on astrocytes, thereby reducing regeneration in the central nervous system.
细胞外基质(ECM)分子在发育过程中和损伤后均参与神经元黏附和神经突生长的调节。我们实验室已证明,星形胶质细胞在细胞外基质分子腱生蛋白的表达上具有异质性。腱生蛋白高表达的星形胶质细胞支持神经突生长的能力降低。此外,用外源性碱性成纤维细胞生长因子(bFGF)处理的星形胶质细胞支持的神经元生长和黏附减少。在本研究中,我们检验了bFGF可增加这些细胞腱生蛋白表达的假说。将碱性成纤维细胞生长因子以高达30 ng/ml的浓度添加到大鼠大脑皮质星形胶质细胞培养物中,该浓度已显示对神经元黏附具有显著影响。通过对细胞提取物和条件培养基进行蛋白质印迹分析以及免疫细胞化学技术来评估腱生蛋白水平。腱生蛋白水平在24 - 48小时后开始增加,并在整个8天的培养过程中持续增加。腱生蛋白的增加呈浓度依赖性,在5 ng/ml bFGF时增加最为显著。在含血清培养基中腱生蛋白产量增加约5.5倍,但在无血清培养基中仅增加约2倍。当在无血清培养基中与bFGF一起加入肝素(10微克/毫升)时,腱生蛋白产量进一步提高。8天后停止bFGF处理,细胞在培养中再维持8天。腱生蛋白水平恢复到对照值,表明bFGF的作用是短暂的。我们的假说是,bFGF在损伤期间的作用可能诱发星形胶质细胞上腱生蛋白的诱导,从而减少中枢神经系统的再生。