Binder L I, Frankfurter A, Kim H, Caceres A, Payne M R, Rebhun L I
Proc Natl Acad Sci U S A. 1984 Sep;81(17):5613-7. doi: 10.1073/pnas.81.17.5613.
The electrophoretic pattern of the large microtubule-associated protein, MAP2, changes during rat brain development. Immunoblots of NaDodSO4 extracts obtained from the cerebral cortex, cerebellum, and thalamus at 10-15 days after birth reveal only a single electrophoretic species when probed with any of three MAP2 monoclonal antibodies. By contrast, adult MAP2 contains two immunoreactive species, MAP2a and MAP2b. The single band of MAP2 from immature brain electrophoretically comigrates with adult MAP2b. Between postnatal days 17 and 18, immature MAP2 simultaneously resolves into two species in both the cerebellum and cerebral cortex. Immunoblots of NaDodSO4 extracts from spinal cord demonstrate the adult complement of MAP2 by day 10, indicating that MAP2 does not change coordinately throughout the entire central nervous system. In vitro cAMP-dependent phosphorylation of immature MAP2 causes a band split reminiscent of that seen during brain development in vivo. The possibility that the developmentally regulated changes observed in MAP2 during brain maturation are due to timed phosphorylation events is discussed.
大鼠脑发育过程中,大型微管相关蛋白MAP2的电泳图谱会发生变化。出生后10 - 15天从大脑皮层、小脑和丘脑获取的十二烷基硫酸钠(NaDodSO4)提取物进行免疫印迹分析,用三种MAP2单克隆抗体中的任何一种进行检测时,均仅显示单一的电泳条带。相比之下,成年MAP2包含两种免疫反应性条带,即MAP2a和MAP2b。未成熟脑的单一MAP2条带在电泳上与成年MAP2b迁移位置相同。在出生后第17天和18天之间,未成熟的MAP2在小脑和大脑皮层中同时分解为两种条带。脊髓NaDodSO4提取物的免疫印迹显示,到第10天时MAP2已具备成年时的组成形式,这表明MAP2在整个中枢神经系统中并非同步变化。未成熟MAP2的体外cAMP依赖性磷酸化导致条带分裂,这与体内脑发育过程中观察到的情况相似。文中讨论了在脑成熟过程中MAP2观察到的发育调控变化可能是由于定时磷酸化事件所致的可能性。