Vogelezang M G, Liu Z, Relvas J B, Raivich G, Scherer S S, ffrench-Constant C
Department of Medical Genetics, University of Cambridge and Cambridge Centre for Brain Repair, University Forvie Site, Cambridge, CB2 2PY, United Kingdom.
J Neurosci. 2001 Sep 1;21(17):6732-44. doi: 10.1523/JNEUROSCI.21-17-06732.2001.
We have shown previously that repair in the peripheral nervous system is associated with a reversion to an embryonic pattern of alternative splicing of the extracellular matrix molecule fibronectin. One of the consequent changes is a relative increase in the number of fibronectins expressing the binding site for alpha4 integrins. Here we show that alpha4 integrins are expressed on dorsal root ganglion neuron cell bodies and growth cones in the sciatic nerve during regeneration and that the interaction of alpha4 integrin with alternatively spliced isoforms of recombinant fibronectins containing the alpha4 binding site enhances neurite outgrowth in dorsal root ganglion neurons. The pheochromocytoma (PC12) neuronal cell line, which normally extends neurites poorly on fibronectin, does so efficiently when alpha4 is expressed in the cells. Experiments using chimeric integrins expressed in PC12 cells show that the alpha4 cytoplasmic domain is necessary and sufficient for this enhanced neurite outgrowth. In both dorsal root ganglion neurons and PC12 cells the alpha4 cytoplasmic domain is tightly linked to the intracellular adapter protein paxillin. These experiments suggest an important role for alpha4 integrin and paxillin in peripheral nerve regeneration and show how alternative splicing of fibronectin may provide a mechanism to enhance repair after injury.
我们之前已经表明,外周神经系统的修复与细胞外基质分子纤连蛋白可变剪接的胚胎模式逆转有关。随之而来的变化之一是表达α4整合素结合位点的纤连蛋白数量相对增加。在此我们表明,α4整合素在再生过程中表达于坐骨神经背根神经节神经元细胞体和生长锥上,并且α4整合素与含有α4结合位点的重组纤连蛋白可变剪接异构体的相互作用增强了背根神经节神经元的神经突生长。嗜铬细胞瘤(PC12)神经元细胞系通常在纤连蛋白上神经突生长较差,但当细胞中表达α4时,它能有效生长。使用在PC12细胞中表达的嵌合整合素进行的实验表明,α4胞质结构域对于这种增强的神经突生长是必要且充分的。在背根神经节神经元和PC12细胞中,α4胞质结构域都与细胞内衔接蛋白桩蛋白紧密相连。这些实验表明α4整合素和桩蛋白在周围神经再生中起重要作用,并展示了纤连蛋白的可变剪接如何可能提供一种增强损伤后修复的机制。