Tucker R P, Hammarback J A, Jenrath D A, Mackie E J, Xu Y
Department of Neurobiology and Anatomy, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157-1010.
J Cell Sci. 1993 Jan;104 ( Pt 1):69-76. doi: 10.1242/jcs.104.1.69.
The glycoprotein tenascin is found in the extracellular matrix in regions of cell motility, cell proliferation, and tissue modelling. We have used novel tenascin cDNA probes to localize tenascin transcripts in the developing mouse and to study the regulation of tenascin expression by growth factors in vitro. At postnatal day 1 tenascin mRNAs are abundant in regions of bone and cartilage formation, as well as in the ependymal layer of the central nervous system. Previous studies have demonstrated that transforming growth factor-beta type 1 (TGF-beta 1) can induce tenascin expression in vitro. As TGF-beta 1 is absent or scarce in the developing brain, it is likely that other growth factors, alone or in addition to TGF-beta 1, may regulate tenascin expression during development. Therefore, we have compared the effects of TGF-beta 1 and a growth factor that is found in both developing connective tissue and the central nervous system, basic fibroblast growth factor (bFGF), on tenascin expression in a mouse embryo fibroblast cell line (Swiss 3T3 cells). Immuno-slot blot analysis of Swiss 3T3 cell-conditioned culture medium demonstrates that bFGF is a more potent inducer of tenascin expression than TGF-beta 1. Furthermore, bFGF and TGF-beta 1 have an additive effect on levels of tenascin, but not fibronectin, in the conditioned medium. Western blots revealed that different forms of tenascin are induced by bFGF and TGF-beta 1: the tenascin induced by the former has a molecular mass of approximately 250 kDa, the latter induces an approximately 200 kDa form of tenascin. The induction of large tenascin by bFGF was confirmed by northern blot analysis, which revealed increased levels of an 8 kb tenascin transcript after 24 h by as little as 4 ng/ml of bFGF in serum-free medium. Thus bFGF, alone or in combination with TGF-beta 1, is a potential regulator of tenascin expression in vitro. bFGF may alter not only the relative abundance of tenascin and fibronectin in the extracellular matrix, but also the splice variant of tenascin expressed by a given cell type.
腱生蛋白这种糖蛋白存在于细胞迁移、细胞增殖和组织重塑区域的细胞外基质中。我们使用了新型的腱生蛋白cDNA探针来定位发育中小鼠体内的腱生蛋白转录本,并在体外研究生长因子对腱生蛋白表达的调控。出生后第1天,腱生蛋白mRNA在骨骼和软骨形成区域以及中枢神经系统的室管膜层中大量存在。先前的研究表明,转化生长因子β1(TGF-β1)在体外可诱导腱生蛋白表达。由于TGF-β1在发育中的大脑中不存在或含量稀少,很可能其他生长因子单独或与TGF-β1一起,在发育过程中调节腱生蛋白的表达。因此,我们比较了TGF-β1和一种在发育中的结缔组织和中枢神经系统中都存在的生长因子——碱性成纤维细胞生长因子(bFGF),对小鼠胚胎成纤维细胞系(瑞士3T3细胞)中腱生蛋白表达的影响。对瑞士3T3细胞条件培养基进行免疫斑点印迹分析表明,bFGF比TGF-β1更有效地诱导腱生蛋白表达。此外,bFGF和TGF-β1对条件培养基中腱生蛋白的水平有相加作用,但对纤连蛋白没有。蛋白质印迹法显示,bFGF和TGF-β1诱导产生不同形式的腱生蛋白:前者诱导产生的腱生蛋白分子量约为250 kDa,后者诱导产生的腱生蛋白约为200 kDa。Northern印迹分析证实了bFGF对大分子量腱生蛋白的诱导作用,该分析显示,在无血清培养基中,低至4 ng/ml的bFGF作用24小时后,8 kb腱生蛋白转录本的水平升高。因此,bFGF单独或与TGF-β1联合,是体外腱生蛋白表达的潜在调节因子。bFGF不仅可能改变细胞外基质中腱生蛋白和纤连蛋白的相对丰度,还可能改变特定细胞类型表达的腱生蛋白剪接变体。