Arregui C, Barra H S
Centro de Investigaciones en Química Biológica de Córdoba, Facultad de Ciencias Quimicas, Universidad Nacional de Córdoba, Argentina.
J Neurosci Res. 1990 Nov;27(3):256-63. doi: 10.1002/jnr.490270303.
We have used the monoclonal YL 1/2 (Tyr antibody) and polyclonal (Glu antibody) antibodies, specific for tyrosinated and detyrosinated tubulin, respectively, to determine the levels and cellular distribution of these tubulin species in chick retina during development. At embryonic day 4, detyrosinated tubulin was restricted to the ganglion cells of the fundic region. As development progresses, immunofluorescence also appears, first, in the outermost zone of the retina and then in the plexiform layers. The Tyr antibody staining was found in the different layers and it was fairly homogeneous in distribution. Analysis by dot immunobinding showed that the ratios of tyrosinated to detyrosinated tubulin obtained at different ages do not agree with those obtained previously by an enzymatic method based on the incorporation of [14C]tyrosine. We found that the lack of coincidence is due to the fact that a fraction of the tubulin species determined by the Tyr and Glu antibodies does not participate in the posttranslational tyrosination/detyrosination cycle. This is a novel concept that should be considered in the interpretations of immunofluorescence studies concerning the cellular distribution of tyrosinated and detyrosinated tubulin.
我们使用了分别针对酪氨酸化微管蛋白和去酪氨酸化微管蛋白的单克隆YL 1/2(酪氨酸抗体)和多克隆(谷氨酸抗体)抗体,来确定这些微管蛋白种类在雏鸡视网膜发育过程中的水平和细胞分布。在胚胎第4天,去酪氨酸化微管蛋白局限于眼底区域的神经节细胞。随着发育的进行,免疫荧光首先出现在视网膜的最外层区域,然后出现在神经纤维层。酪氨酸抗体染色在不同层均有发现,且分布相当均匀。斑点免疫结合分析表明,不同年龄获得的酪氨酸化与去酪氨酸化微管蛋白的比例与先前基于[14C]酪氨酸掺入的酶法获得的比例不一致。我们发现这种不一致是由于酪氨酸和谷氨酸抗体所确定的一部分微管蛋白种类不参与翻译后酪氨酸化/去酪氨酸化循环。这是一个新的概念,在解释关于酪氨酸化和去酪氨酸化微管蛋白细胞分布的免疫荧光研究时应予以考虑。