Falconer M M, Vielkind U, Brown D L
University of Ottawa, Department of Biology, Ont., Canada.
Biochem Cell Biol. 1989 Sep;67(9):537-44. doi: 10.1139/o89-085.
Pluripotent P19 embryonal carcinoma cell cultures can be induced to differentiate into neurons and glial cells by the addition of 10(-6) M retinoic acid. During early neural differentiation, a bundle of colchicine-stable, acetylated microtubules is formed. This acetylated microtubule array apparently extends to form neurites during neurogenesis. In this paper, we analyze changes in vimentin and MAP 2 distributions during neural differentiation with respect to the changes in the acetylated microtubule array. During a brief period early in differentiation, indirect immunofluorescence staining shows the colocalization of colchicine-stable acetylated microtubules, vimentin, and MAP 2. Using acrylamide to disrupt the organization of vimentin intermediate filaments and estramustine to disrupt the binding of MAP 2 to microtubules, we show that acetylated microtubules, MAP 2, and vimentin intermediate filaments are arranged in an interdependent cytoskeletal array. We suggest this array may serve to stabilize processes in neural stem cells, before the final decision to differentiate into neurons or glia is made.
通过添加10⁻⁶ M视黄酸，多能性P19胚胎癌细胞培养物可被诱导分化为神经元和神经胶质细胞。在早期神经分化过程中，会形成一束对秋水仙碱稳定的乙酰化微管。这种乙酰化微管阵列显然在神经发生过程中延伸形成神经突。在本文中，我们分析了神经分化过程中波形蛋白和微管相关蛋白2（MAP 2）分布的变化与乙酰化微管阵列变化的关系。在分化早期的一段短暂时间内，间接免疫荧光染色显示秋水仙碱稳定的乙酰化微管、波形蛋白和MAP 2共定位。使用丙烯酰胺破坏波形蛋白中间丝的组织，并使用雌莫司汀破坏MAP 2与微管的结合，我们表明乙酰化微管、MAP 2和波形蛋白中间丝排列成相互依赖的细胞骨架阵列。我们认为，在最终决定分化为神经元或神经胶质细胞之前，这种阵列可能有助于稳定神经干细胞中的突起。
