Merrick S E, Pleasure S J, Lurie D I, Pijak D S, Selzer M E, Lee V M
David Mahoney Institute of Neurological Sciences, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA.
J Comp Neurol. 1995 May 1;355(2):199-210. doi: 10.1002/cne.903550204.
Lamprey axons regenerate following spinal cord transection despite the formation of a glial scar. As we were unable to detect a lamprey homologue of glial fibrillary acidic protein (GFAP), a major constituent of astrocytes, we studied the composition of intermediate filament (IF) proteins of lamprey glia. Monoclonal antibodies (mAbs) were raised to lamprey spinal cord cytoskeletal extracts and these mAbs were characterized by using Western blotting and immunocytochemistry. On two-dimensional (2-D) Western blots, five of the mAbs detected three major IF polypeptides in the molecular weight (MW) range of 45-56 kD. Further studies were conducted to determine the relationship between the lamprey glial-specific antigen and other mammalian IF proteins. Antikeratin 8 antibody recognized two of the three polypeptides. Several of the glial-specific mAbs reacted with human keratins 8 and 18 on Western blots. Keratin-like immunoreactivity was found in all parts of the central and peripheral nervous systems in both larval and adult lampreys. The immunocytochemical staining patterns of glial-specific mAbs were indistinguishable on lamprey spinal cord sections. However, on brain sections, two distinct patterns were observed. A subset of mAbs stained only a few glial fibers in the brain, whereas others stained many more brain glia, particularly the ependymal cells. The former group of mAbs recognized only the two lower MW polypeptides on 2-D Western blots, but the latter group of mAbs recognized all three major IF polypeptides. This correlation is supported by the observation that the highest MW IF polypeptide has an increased level of expression in the brain relative to the spinal cord. Thus, in the lamprey, the glial cells of both spinal cord and brain express molecules similar to simple epithelial cytokeratins, but their IFs may contain these keratins in different stoichiometric proportions. The widespread presence in the lamprey of primitive glial cells containing keratin-like intermediate filaments may have significance for the extraordinary ability of lamprey spinal axons to regenerate.
七鳃鳗的轴突在脊髓横断后仍能再生,尽管会形成胶质瘢痕。由于我们无法检测到作为星形胶质细胞主要成分的胶质纤维酸性蛋白(GFAP)的七鳃鳗同源物,因此我们研究了七鳃鳗胶质细胞中间丝(IF)蛋白的组成。制备了针对七鳃鳗脊髓细胞骨架提取物的单克隆抗体(mAb),并通过蛋白质印迹法和免疫细胞化学对这些mAb进行了表征。在二维(2-D)蛋白质印迹上,其中5种mAb在分子量(MW)为45 - 56 kD范围内检测到3种主要的IF多肽。进行了进一步的研究以确定七鳃鳗胶质细胞特异性抗原与其他哺乳动物IF蛋白之间的关系。抗角蛋白8抗体识别出这3种多肽中的2种。几种胶质细胞特异性mAb在蛋白质印迹上与人角蛋白8和18发生反应。在幼虫和成年七鳃鳗的中枢神经系统和外周神经系统的所有部位都发现了角蛋白样免疫反应性。胶质细胞特异性mAb在七鳃鳗脊髓切片上的免疫细胞化学染色模式无法区分。然而,在脑切片上,观察到两种不同的模式。一部分mAb仅对脑中的少数胶质纤维染色,而其他mAb则对更多的脑胶质细胞染色,特别是室管膜细胞。前一组mAb在二维蛋白质印迹上仅识别两种较低MW的多肽,但后一组mAb识别所有三种主要的IF多肽。这种相关性得到了以下观察结果的支持:相对于脊髓,最高MW的IF多肽在脑中的表达水平有所增加。因此,在七鳃鳗中,脊髓和脑的胶质细胞都表达类似于简单上皮细胞角蛋白的分子,但它们的IFs可能以不同的化学计量比例包含这些角蛋白。在七鳃鳗中广泛存在含有角蛋白样中间丝的原始胶质细胞可能对七鳃鳗脊髓轴突非凡的再生能力具有重要意义。