Knight R M, Fossom L H, Neuberger T J, Attema B L, Tennekoon G, Bharucha V, DeVries G H
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Richmond 23298-0614.
J Neurosci Res. 1993 May 1;35(1):38-45. doi: 10.1002/jnr.490350106.
To elucidate the role of axonal plasma membrane factors in the differentiation of Schwann cells, we investigated the effect of an axolemma-enriched fraction (AEF) isolated from myelinated CNS tissue on the expression of P0 glycoprotein, the major glycoprotein in peripheral myelin, in primary rat Schwann cells (PSC) isolated from sciatic nerve, as well as in a transfected rat Schwann cell line (TSC). AEF increased PO-mRNA levels in PSC and TSC in a concentration-dependent manner, producing a maximal induction of nearly twofold after 48 hr of treatment. A similar induction of P0 mRNA was elicited in TSC by the cAMP-activating agents 8-bromo-cAMP and forskolin, which have been shown to induce myelin proteins in PSC. In addition to inducing P0 mRNA, AEF and forskolin also increased the amount of P0 protein in TSC, as indicated by increased P0-immunoreactive staining. However, in TSC, axolemma caused no increase in expression of CAT linked to a P0 promoter while forskolin caused a marked increase in the expression from the P0 promoter. These results suggest that AEF, in contrast to forskolin, does not regulate P0-mRNA expression at the level of transcriptional activity. These in vitro systems may be useful for the study of axolemmal factors that induce Schwann cell differentiation.
为阐明轴突质膜因子在雪旺细胞分化中的作用,我们研究了从有髓中枢神经系统组织中分离出的富含轴膜的组分(AEF)对从坐骨神经分离出的原代大鼠雪旺细胞(PSC)以及转染的大鼠雪旺细胞系(TSC)中外周髓鞘主要糖蛋白P0糖蛋白表达的影响。AEF以浓度依赖的方式增加PSC和TSC中P0 - mRNA水平,处理48小时后产生近两倍的最大诱导作用。cAMP激活剂8 - 溴 - cAMP和福斯可林在TSC中也引发了类似的P0 mRNA诱导,这两种物质已被证明可在PSC中诱导髓鞘蛋白。除了诱导P0 mRNA外,AEF和福斯可林还增加了TSC中P0蛋白的量,P0免疫反应性染色增加表明了这一点。然而,在TSC中,轴膜并未导致与P0启动子相连的CAT表达增加,而福斯可林则导致P0启动子的表达显著增加。这些结果表明,与福斯可林不同,AEF在转录活性水平上不调节P0 - mRNA表达。这些体外系统可能有助于研究诱导雪旺细胞分化的轴膜因子。