Matthieu J M, Waehneldt T V, Webster H D, Bény M, Fagg G E
Brain Res. 1979 Jul 6;170(1):123-33. doi: 10.1016/0006-8993(79)90945-4.
Myelin was purified from adult rabbit sciatic nerve by two procedures: discontinuous gradient centrifugation and continuous gradient zonal centrifugation. Two fractions were obtained from the discontinuous gradient. The fraction floating on 0.32 M sucrose and the fraction recovered from the 0.32/0.85 M sucrose interface showed typical myelin membranes by electron microscopy and typical myelin proteins by gel electrophoresis. The specific activity of 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) decreased from the top to the bottom of the discontinuous gradient. The myelin separated by zonal centrifugation on a continuous sucrose gradient showed three distinct peaks (on monitoring optical density) at 0.10, 0.30 and 0.57 M sucrose. The latter peak yielded 92% of the material applied. The two minor peaks of low density exhibited high CNP and acetylcholinesterase (AChE) activities but the specific activity of both enzymes increased markedly at the heavy end of the gradient. The zonal fractions showed typical myelin proteins in all fractions by polyacrylamide gel electrophoresis but with important quantitative differences. These results indicate that PNS myelin shows significant heterogeneity.
不连续梯度离心和连续梯度区带离心。从不连续梯度中获得了两个组分。漂浮在0.32M蔗糖上的组分以及从0.32/0.85M蔗糖界面回收的组分，通过电子显微镜显示出典型的髓鞘膜，通过凝胶电泳显示出典型的髓鞘蛋白。2',3'-环核苷酸3'-磷酸二酯酶（CNP）的比活性从不连续梯度的顶部到底部逐渐降低。在连续蔗糖梯度上通过区带离心分离的髓磷脂在0.10、0.30和0.57M蔗糖处显示出三个不同的峰（通过监测光密度）。后一个峰产生了92%的上样物质。两个低密度的小峰表现出高CNP和乙酰胆碱酯酶（AChE）活性，但两种酶的比活性在梯度的重端显著增加。通过聚丙烯酰胺凝胶电泳，区带组分在所有组分中都显示出典型的髓鞘蛋白，但存在重要的定量差异。这些结果表明，外周神经系统髓磷脂表现出显著的异质性。
