Mikoshiba K, Takamatsu K, Tsukada Y
Brain Res. 1983 Mar;283(1):71-9. doi: 10.1016/0165-3806(83)90082-2.
Peripheral nerves of the shiverer mouse, which are characterized by the absence of major dense lines and myelin basic proteins in CNS myelin, were analyzed. From subcellular fractionation of sciatic nerves, it was found from the SDS-polyacrylamide gel electrophoresis that the Pl and Pr proteins equivalent to myelin basic protein of CNS and PM protein were missing in the shiverer in both P2A and P3A fractions in which PNS myelin is recovered. No extra bands were observed in any other fractions of the shiverer in place of the absence of the proteins. The activities of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) of P2A and P3A fractions were high, but that of the P3X fraction which floated over 0.32 M sucrose was the highest among the fractions examined in both the shiverer and the control. Developmental analysis of the protein profiles revealed that PO, Pl, Pr and PM proteins increased rapidly from the sixth day postnatally up to the twentieth day after birth in the control. No differences were observed between the shiverer and the control as for PO protein, but Pl, Pr and PM proteins were absent in the shiverer throughout the development. The CNPase activity of total homogenate of sciatic nerve fibers at birth in the control showed high activity comparable to that of the adult value, but there was no significant difference in activity between the control and the shiverer at any stage of development. Immunohistochemical reaction using peroxidase anti-peroxidase method showed that the myelin from the shiverer did not react with the MBP antiserum, while that of the control reacted positively. On the contrary, the myelin from both shiverer and the control reacted positively against P2 antibody.
对颤抖鼠的外周神经进行了分析,其特征是中枢神经系统髓鞘中缺乏主要致密线和髓鞘碱性蛋白。通过对坐骨神经进行亚细胞分级分离,从十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中发现,在回收外周神经系统髓鞘的P2A和P3A组分中,与中枢神经系统髓鞘碱性蛋白相当的Pl和Pr蛋白以及外周髓鞘蛋白(PM蛋白)在颤抖鼠中缺失。在颤抖鼠的任何其他组分中均未观察到替代这些缺失蛋白的额外条带。P2A和P3A组分的2',3'-环核苷酸3'-磷酸水解酶(CNPase)活性较高,但在0.32M蔗糖上浮的P3X组分的活性在颤抖鼠和对照所检测的组分中是最高的。蛋白质谱的发育分析表明,在对照中,PO、Pl、Pr和PM蛋白从出生后第六天到出生后第二十天迅速增加。在PO蛋白方面,颤抖鼠和对照之间未观察到差异,但在整个发育过程中,颤抖鼠中Pl、Pr和PM蛋白均缺失。对照中出生时坐骨神经纤维总匀浆的CNPase活性显示出与成年值相当的高活性,但在发育的任何阶段,对照和颤抖鼠之间的活性均无显著差异。使用过氧化物酶抗过氧化物酶方法的免疫组织化学反应表明,颤抖鼠的髓鞘不与髓鞘碱性蛋白抗血清反应,而对照的髓鞘则呈阳性反应。相反,颤抖鼠和对照的髓鞘对P2抗体均呈阳性反应。