Kaplan M P, Chin S S, Fliegner K H, Liem R K
Department of Pathology, Columbia University College of Physicians and Surgeons, New York, New York 10032.
J Neurosci. 1990 Aug;10(8):2735-48. doi: 10.1523/JNEUROSCI.10-08-02735.1990.
alpha-Internexin is a 66 kDa protein that copurifies with intermediate filaments (IF) from rat spinal cord and optic nerve. This protein is axonally transported in rat optic nerve along with the neurofilament triplet proteins in slow component a. Polymerization in vitro and distribution in vivo confirm that alpha-internexin is a neuronal IF. We raised 2 highly specific monoclonal antibodies to alpha-internexin which were applied to frozen rat brain sections and Western blots of cytoskeletal extracts. These results indicate that alpha-internexin is primarily an axonal protein found in most, if not all, neurons of the CNS. Immunoreactive proteins of similar molecular weight were found in cytoskeletal extracts of CNS tissue from several additional species, including mouse and cow. While the distribution of alpha-internexin as given by immunocytochemical methods is similar to that of low molecular weight neurofilament protein (NF-L) in the adult, its distribution in the embryo is far more extensive. At embryonic day 16, when the expression of NF-L is still limited to a relatively small number of cells and levels of expression are low, alpha-internexin is already found at much higher levels and in cells not yet expressing NF-L in detectable quantities. Similar results are found at embryonic day 12. These data suggest that neuronal IF in the developing nervous system contain a higher proportion of alpha-internexin than their adult counterparts, and that expression of alpha-internexin precedes that of NF-L in many or most neurons of the developing brain.
α-中间丝蛋白是一种66 kDa的蛋白质,它与来自大鼠脊髓和视神经的中间丝(IF)共同纯化。这种蛋白质在大鼠视神经中与神经丝三联体蛋白一起在慢运输组分a中进行轴突运输。体外聚合和体内分布证实α-中间丝蛋白是一种神经元中间丝。我们制备了两种高度特异的针对α-中间丝蛋白的单克隆抗体,并将其应用于大鼠脑冰冻切片和细胞骨架提取物的蛋白质免疫印迹分析。这些结果表明,α-中间丝蛋白主要是一种轴突蛋白,存在于中枢神经系统的大多数(如果不是全部)神经元中。在包括小鼠和牛在内的其他几种物种的中枢神经系统组织的细胞骨架提取物中也发现了分子量相似的免疫反应性蛋白。虽然免疫细胞化学方法显示的α-中间丝蛋白的分布在成体中与低分子量神经丝蛋白(NF-L)相似,但其在胚胎中的分布要广泛得多。在胚胎第16天,当NF-L的表达仍局限于相对少数的细胞且表达水平较低时,α-中间丝蛋白已经在更高水平上被发现,并且在尚未检测到可检测量NF-L的细胞中也有表达。在胚胎第12天也发现了类似的结果。这些数据表明,发育中的神经系统中的神经元中间丝比成体中的含有更高比例的α-中间丝蛋白,并且在发育中的大脑的许多或大多数神经元中,α-中间丝蛋白的表达先于NF-L。