Velasco A, Bragado M J, Jimeno D, Caminos E, Lillo C, Aijón J, Lara J M
Instituto de Neurociencias de Castilla y León, Departamento de Biología Celular y Patología, Universidad de Salamanca, E-37007, Salamanca, Spain.
Brain Res. 2000 Nov 10;883(1):98-106. doi: 10.1016/s0006-8993(00)02783-9.
We have analyzed the immunolabeling with the antibody RT97, a good marker for ganglion cell axons in several species, in the normal and regenerating visual pathways of teleosts. We have demonstrated that RT97 antibody recognizes several proteins in the tench visual system tissues (105, 115, 160, 200, 325 and 335 kDa approximately). By using immunoprecipitation and Western blot we have found that after crushing the optic nerve the immunoreactivity to anti RT97 increased markedly in the optic nerve. In immunohistochemical analysis we also found a different pattern of labeling in normal and regenerating visual pathways. In normal tench RT97 is a good marker for the horizontal cells in the retina, for growing ganglion cell axons which run along the optic nerve from the retina to the optic tectum and of the axon terminals in the stratum opticum and stratum fibrosum and griseum superficiale in the optic tectum. After optic nerve crush, no immunohistochemistry modifications were observed in the retina. However, in accordance with Western blot experiments, in the optic nerve intensely stained groups of regenerating axons appeared progressively throughout the optic nerve as far as the optic tectum. We conclude that the antibody RT97 is an excellent marker of growing and regenerating axons of the optic nerve of fish.
我们使用抗体RT97（在多个物种中是神经节细胞轴突的良好标志物）对硬骨鱼正常和再生视觉通路进行了免疫标记分析。我们已经证明，RT97抗体可识别丁鱥视觉系统组织中的几种蛋白质（大约为105、115、160、200、325和335 kDa）。通过免疫沉淀和蛋白质印迹法，我们发现视神经挤压后，视神经中抗RT97的免疫反应性显著增加。在免疫组织化学分析中，我们还发现正常和再生视觉通路中的标记模式不同。在正常丁鱥中，RT97是视网膜水平细胞、从视网膜沿视神经向视顶盖生长的神经节细胞轴突以及视顶盖视层、纤维层和浅灰层中轴突终末的良好标志物。视神经挤压后，视网膜未观察到免疫组织化学变化。然而，与蛋白质印迹实验一致，在视神经中，再生轴突的强染色组逐渐出现在整个视神经直至视顶盖。我们得出结论，抗体RT97是鱼类视神经生长和再生轴突的优秀标志物。
