Ruiz-Ederra Javier, García Monica, Hicks David, Vecino Elena
Universidad del País Vasco, Facultad de Medicina, Departamento Biología Celular, Leioa, Vizcaya, Spain.
Mol Vis. 2004 Feb 10;10:83-92.
Neurofilaments (NF) are neuronal cytoskeletal components and immunostaining against them has been used to visualize retinal ganglion cells (RGC) and their axons. Since the RGC cytoskeleton exhibits differential damage in diseases such as glaucoma, we examined the distribution of light, medium, and heavy NF subunits (NF-L, NF-M, and NF-H respectively) within normal human and porcine retinas, as a function of RGC soma size and eccentricity.
NF subunits were visualized with immunofluorescence techniques using retinal sections and flatmounts from adult human and pig retinas that were incubated with specific antisera against the three NF subunits. Porcine RGCs were retrogradely labeled with fluorogold while human RGCs were identified based on their position within the inner retina and their relatively large somata.
NF-H and NF-M were distributed widely within all RGC somata and dendrites, whereas NF-L was more restricted to the perinuclear area. In addition, phosphorylated NF-H distribution varied with retinal eccentricity so a subpopulation of large RGCs located in the peripheral retina was intensely labeled with the antiserum recognizing the phosphorylated NF-H.
We show that at least one of each of three NF subunits is present in all RGCs in porcine and presumably in human retina, and that NF distribution is very similar in RGCs of both species.
神经丝（NF）是神经元细胞骨架成分，针对它们的免疫染色已被用于可视化视网膜神经节细胞（RGC）及其轴突。由于RGC细胞骨架在青光眼等疾病中表现出不同程度的损伤，我们研究了正常人和猪视网膜中轻、中、重NF亚基（分别为NF-L、NF-M和NF-H）的分布，作为RGC胞体大小和离心率的函数。
使用免疫荧光技术对成人和猪视网膜的切片及平铺片进行NF亚基可视化，这些切片和平铺片用针对三种NF亚基的特异性抗血清孵育。猪RGC用荧光金逆行标记，而人RGC根据其在内视网膜中的位置及其相对较大的胞体来识别。
NF-H和NF-M广泛分布于所有RGC胞体和树突中，而NF-L更多地局限于核周区域。此外，磷酸化NF-H的分布随视网膜离心率而变化，因此位于周边视网膜的一大群大RGC被识别磷酸化NF-H的抗血清强烈标记。
我们表明，猪视网膜以及推测人视网膜中的所有RGC中至少各存在三种NF亚基中的一种，并且两种物种的RGC中NF分布非常相似。
