Parysek L M, McReynolds M A, Goldman R D, Ley C A
University of Cincinnati, Department of Anatomy and Cell Biology, Ohio 45267-0521.
J Neurosci Res. 1991 Sep;30(1):80-91. doi: 10.1002/jnr.490300110.
Mammalian neurons and neuron-like cultured cells express the neural intermediate filament (IF) proteins neurofilament (NF)-L, NF-M, NF-H, and peripherin. To determine whether these proteins are found within the same 10-nm filament, light and electron microscope immunocytochemistry using peripherin and NF-specific antibodies was performed on PC12 cells, nervous tissue, and isolated neural filaments from the cauda equina. Double-label immunofluorescence showed that peripherin and NF-L, -M, and -H were found in identical filamentous patterns in interphase and mitotic PC12 cells. Furthermore, expression of mutant peripherin in PC12 cells disrupted not only the peripherin network but also NF-containing filaments. Immunoelectron microscopy of PC12 cell cytoskeletons showed that peripherin and NF subunit proteins were found in the same filament. In situ, in the sciatic nerve, peripherin/NF-L or peripherin/NF-M/-H double-label immunofluorescence illustrates at least three types of nerve fibers: those containing NF only, those labeled predominantly for peripherin, and fibers in which peripherin and NF subunits were colocalized. Immunoelectron microscopy of filaments isolated from nerve roots comprising the sciatic nerve also showed the same three labeling patterns seen by light microscopy. Some neural IF appear to contain predominantly NF proteins or peripherin, but in others, both proteins are found within the same IF.
哺乳动物神经元和类神经元培养细胞表达神经中间丝（IF）蛋白神经丝（NF）-L、NF-M、NF-H和外周蛋白。为了确定这些蛋白质是否存在于同一10纳米丝中，使用外周蛋白和NF特异性抗体对PC12细胞、神经组织和马尾神经分离的神经丝进行了光镜和电镜免疫细胞化学研究。双重标记免疫荧光显示，外周蛋白和NF-L、-M、-H在间期和有丝分裂期的PC12细胞中呈现相同的丝状模式。此外，PC12细胞中外周蛋白突变体的表达不仅破坏了外周蛋白网络，还破坏了含NF的丝。PC12细胞细胞骨架的免疫电子显微镜显示，外周蛋白和NF亚基蛋白存在于同一丝中。在坐骨神经原位，外周蛋白/NF-L或外周蛋白/NF-M/-H双重标记免疫荧光显示至少三种类型的神经纤维：仅含NF的神经纤维、主要标记外周蛋白的神经纤维以及外周蛋白和NF亚基共定位的纤维。从构成坐骨神经的神经根分离的丝的免疫电子显微镜也显示了光镜下所见的相同三种标记模式。一些神经中间丝似乎主要含有NF蛋白或外周蛋白，但在其他丝中，两种蛋白都存在于同一中间丝中。
