Li Wen, Xia Jin-tang, Feng Yue
Laboratory of Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
Acta Pharmacol Sin. 2006 Sep;27(9):1119-26. doi: 10.1111/j.1745-7254.2006.00362.x.
To study the role of microtubule dynamics and microtubule associated protein 1B (MAP1B) in regulation of the neurite extension in CAD catecholaminergic neuronal cell line.
The neuritogenesis of the CAD cells was abolished by inhibiting microtubule polymerization with nocodazole and by blocking microtubule depolymerization with taxol. MAP1B and tubulin protein expression was detected by Western blot. Immunofluorescent staining of tubulins was observed by fluorescent and confocal microscopy.
Microtubule dynamics was essential for CAD neurite extension. Dosage analysis revealed that neurite extension was much more sensitive to nocodazole than to taxol, suggesting a functional requirement for highly active microtubule assembly. A remarkable upregulation of MAP1B protein was detected during neurite extension accompanied with increased microtubule stability.
Upregulation of MAP1B leads to the stabilization of newly formed microtubules in the developing neurites, which in turn promotes neurite extension.
研究微管动力学及微管相关蛋白1B(MAP1B)在调节CAD儿茶酚胺能神经元细胞系神经突延伸中的作用。
用诺考达唑抑制微管聚合以及用紫杉醇阻断微管解聚,从而消除CAD细胞的神经突形成。通过蛋白质免疫印迹法检测MAP1B和微管蛋白的表达。用荧光显微镜和共聚焦显微镜观察微管蛋白的免疫荧光染色。
微管动力学对CAD神经突延伸至关重要。剂量分析显示,神经突延伸对诺考达唑比对紫杉醇更敏感,提示对高活性微管组装有功能需求。在神经突延伸过程中检测到MAP1B蛋白显著上调,同时微管稳定性增加。
MAP1B的上调导致发育中的神经突中新形成的微管稳定,进而促进神经突延伸。