Hopkins J M, Ford-Holevinski T S, McCoy J P, Agranoff B W
J Neurosci. 1985 Nov;5(11):3030-8. doi: 10.1523/JNEUROSCI.05-11-03030.1985.
Previous work from our laboratory had shown that goldfish retinal fragments explanted onto a polylysine substratum 1 to 2 weeks following optic nerve crush exhibit a striking clockwise pattern of neuritic outgrowth. In the present study, however, when the basal lamina component laminin was used as a substratum, neurites grew out as uncurved spokes, were less fasciculated, and had an increased rate of elongation. When laminin was combined with polylysine in the substratum, the degree of fasciculation and rate of elongation were similar to those seen on laminin alone, whereas the tendency for clockwise outgrowth was even more pronounced than that observed with polylysine alone. These results suggest that regenerating neurites have an affinity for laminin. Using an antibody to murine Engelbreth-Holm-Swarm sarcoma laminin, which cross-reacted with basal lamina in goldfish tissue sections, we studied the histochemical distribution of laminin in the goldfish visual system. Immunoperoxidase staining for laminin showed a characteristic scalloped pattern of staining in cross-sections of optic nerve bundles. Following optic nerve crush, the reaction product became much more diffuse and intense, especially distal to the crush site. When the retinal ganglion cell bodies were eliminated by removing the eye, the degenerating optic nerve stump still showed the intensive staining. We interpret these results to indicate that optic nerve glia are responsible in large part for the formation of laminin. Taken together, these in vivo and in vitro findings suggest that laminin plays a role in nerve regeneration in the goldfish central nervous system.
我们实验室之前的研究表明,视神经挤压伤后1至2周,移植到聚赖氨酸基质上的金鱼视网膜碎片呈现出明显的顺时针神经突生长模式。然而,在本研究中,当将基底膜成分层粘连蛋白用作基质时,神经突呈直线状长出,束状化程度较低,伸长率增加。当层粘连蛋白与聚赖氨酸在基质中结合时,束状化程度和伸长率与单独使用层粘连蛋白时相似,而顺时针生长的趋势比单独使用聚赖氨酸时更为明显。这些结果表明,再生神经突对层粘连蛋白具有亲和力。我们使用一种与金鱼组织切片中的基底膜发生交叉反应的抗小鼠恩格尔布雷特-霍尔姆-斯旺肉瘤层粘连蛋白抗体,研究了层粘连蛋白在金鱼视觉系统中的组织化学分布。层粘连蛋白的免疫过氧化物酶染色在视神经束横切面上显示出特征性的扇贝状染色模式。视神经挤压伤后,反应产物变得更加弥散和强烈,尤其是在挤压部位的远端。当通过摘除眼球消除视网膜神经节细胞体时,变性的视神经残端仍显示出强烈的染色。我们将这些结果解释为表明视神经胶质细胞在很大程度上负责层粘连蛋白的形成。综上所述,这些体内和体外研究结果表明层粘连蛋白在金鱼中枢神经系统的神经再生中发挥作用。