Ma D, Chow S, Obrocka M, Connors T, Fischer I
Department of Neurobiology and Anatomy, Medical College of Pennsylvania Hahnemann University, 3200 Henry Avenue, Philadelphia, PA 19129, USA.
Brain Res. 1999 Mar 27;823(1-2):141-53. doi: 10.1016/s0006-8993(99)01148-8.
Microtubule-associated protein 1B (MAP1B) is expressed at high levels during development of the nervous system and is localized primarily in neurons while specific phosphorylated isoforms of MAP1B are localized exclusively in growing axons. The levels of MAP1B are down regulated in most regions of the adult CNS, but remain high in neurons and axons of the PNS. This study demonstrates that the expression of MAP1B is induced in adult Schwann cells following sciatic nerve lesion and regeneration. High levels of both mRNA and the MAP1B protein were detected in Schwann cells associated with the axotomized distal stump. Expression of MAP1B was also observed in cultured primary Schwann cells from neonatal rats. The properties of the MAP1B protein in cultured Schwann cells were further characterized by Western blot analysis using specific antibodies that recognize the N-terminal, middle and C-terminal domains of MAP1B. All of these antibodies detected a protein of 320-340 kDa demonstrating that MAP1B expressed by Schwann cells is very similar, or identical, to MAP1B expressed by neurons. The phosphorylation of MAP1B in Schwann cells was also studied using monoclonal antibodies (mAb) that recognize specific phosphorylation epitopes. The results indicated that the expression of MAP1B in Schwann cells exhibited a differential phosphorylation state that was recognized by mAb 1B6 but not by other mAbs, including 1B-P, 150 and RT97, that recognize phosphorylated MAP1B in growing axons. We therefore conclude that MAP1B is expressed in Schwann cells during both development and axonal regeneration, suggesting that the developmental pattern of MAP1B in these cells is recapitulated in adult Schwann cells during the early stages of regeneration and remyelination of injured peripheral axons. The presence of MAP1B in Schwann cells may support morphological changes of these cells, particularly the formation of processes prior to their differentiation into myelin forming Schwann cells.
微管相关蛋白1B(MAP1B)在神经系统发育过程中高水平表达,主要定位于神经元,而MAP1B的特定磷酸化异构体则仅定位于生长中的轴突。在成体中枢神经系统的大多数区域,MAP1B水平下调,但在周围神经系统的神经元和轴突中仍保持较高水平。本研究表明,坐骨神经损伤和再生后,成体雪旺细胞中MAP1B的表达被诱导。在与轴突切断的远端残端相关的雪旺细胞中检测到高水平的mRNA和MAP1B蛋白。在新生大鼠的原代培养雪旺细胞中也观察到了MAP1B的表达。使用识别MAP1B N端、中间和C端结构域的特异性抗体,通过蛋白质印迹分析进一步表征了培养的雪旺细胞中MAP1B蛋白的特性。所有这些抗体都检测到一种320 - 340 kDa的蛋白质,表明雪旺细胞表达的MAP1B与神经元表达的MAP1B非常相似或相同。还使用识别特定磷酸化表位的单克隆抗体(mAb)研究了雪旺细胞中MAP1B的磷酸化情况。结果表明,雪旺细胞中MAP1B的表达呈现出一种差异磷酸化状态,可被mAb 1B6识别,但不被其他识别生长轴突中磷酸化MAP1B的mAb识别,包括1B - P、150和RT97。因此,我们得出结论,MAP1B在发育和轴突再生过程中均在雪旺细胞中表达,这表明在成年雪旺细胞中,在受损周围轴突再生和髓鞘再生的早期阶段,这些细胞中MAP1B的发育模式得以重现。雪旺细胞中MAP1B的存在可能支持这些细胞的形态变化,特别是在它们分化为形成髓鞘的雪旺细胞之前突起的形成。