Bates C A, Trinh N, Meyer R L
Developmental Biology Center, University of California, Irvine 92717.
Dev Biol. 1993 Feb;155(2):533-44. doi: 10.1006/dbio.1993.1050.
Microtubule-associated proteins (MAPs) are developmentally regulated proteins involved in microtubule polymerization and stabilization. We have asked whether MAPs in regenerating adult mouse retinal ganglion cell axons recapitulate the pattern of MAPs seen during development. Adult and Embryonic Day 15 (E15) mouse retinal segments were explanted onto laminin-coated coverslips and allowed to extend neurites in serum-free medium. Using a variety of monoclonal antibodies (Mabs), we stained adult and E15 explant cultures for early (MAP5) and late (MAP1, MAP2, MAP2a+b, tau) MAPs. Optic neurites growing from E15 explants were strongly immunoreactive for MAP5. Labeling was absent for MAP1 and MAP2a+b. Mabs to MAP2 and tau lightly stained axons and cell bodies, probably due to MAP2c and juvenile tau which are found in developing neurons. In adult explant cultures, both perikarya and axons were immunoreactive for MAP1. MAP2 and MAP2a+b labeled perikarya and fibers within the explant, and tau stained axons on the substrate. MAP5 immunoreactivity was intense in the adult explant cultures, labeling axons extending over the substrate and perikarya and dendrites in the explant in a pattern similar to that seen in E15 explants. In summary, adult retinal explants maintain their adult complement of MAPs, in showing high levels of MAP1, tau immunoreactivity restricted to neurites on the substrate, and MAP2 segregated into neurites and perikarya within the explant. The exception is MAP5 for which the pattern of immunoreactivity is similar in adult and E15 retinal explants.
微管相关蛋白(MAPs)是参与微管聚合和稳定的发育调控蛋白。我们研究了成年小鼠视网膜神经节细胞轴突再生过程中的MAPs是否重现发育过程中所见的MAPs模式。将成年和胚胎第15天(E15)小鼠的视网膜节段接种到层粘连蛋白包被的盖玻片上,并在无血清培养基中使其长出神经突。使用多种单克隆抗体(Mabs),我们对成年和E15外植体培养物进行染色,以检测早期(MAP5)和晚期(MAP1、MAP2、MAP2a + b、tau)MAPs。从E15外植体长出的视神经突对MAP5有强烈的免疫反应。MAP1和MAP2a + b无标记。针对MAP2和tau的单克隆抗体对轴突和细胞体有轻度染色,这可能是由于发育中的神经元中存在的MAP2c和幼年tau。在成年外植体培养物中,核周体和轴突对MAP1均有免疫反应。MAP2和MAP2a + b标记了外植体内的核周体和纤维,tau则对底物上的轴突进行了染色。在成年外植体培养物中,MAP5免疫反应强烈,其标记底物上延伸的轴突以及外植体内的核周体和树突,其模式与E15外植体中所见相似。总之,成年视网膜外植体保持其成年MAPs的组成,表现为高水平的MAP1、tau免疫反应仅限于底物上的神经突,而MAP2则在外植体内的神经突和核周体中分离。例外的是MAP5,其免疫反应模式在成年和E15视网膜外植体中相似。