Larocca Jorge N, Norton Williams T
Albert Einstein College of Medicine, Yeshiva University, Bronx, New York, USA.
Curr Protoc Cell Biol. 2007 Jan;Chapter 3:Unit3.25. doi: 10.1002/0471143030.cb0325s33.
The methods used to prepare myelin involve homogenization of the tissue in isotonic sucrose solution, followed by the isolation of myelin membranes by a series of steps that include density gradient centrifugation and differential centrifugation. Homogenization of nervous tissue in isotonic sucrose causes the myelin sheath to peel from the axon and form relatively large myelin vesicles. The large size of the myelin vesicles, together with the fact that myelin membrane has a lower density than other biological membranes, make differential centrifugation and density gradient centrifugation the main tools for the isolation of this membrane. Three protocols are outlined in this unit: isolation of a highly-purified myelin fraction from the central nervous system (CNS); separation of a highly-purified CNS myelin fraction into subfractions of different densities; and isolation of myelin from the peripheral nervous system (PNS).
制备髓磷脂的方法包括将组织在等渗蔗糖溶液中匀浆，随后通过一系列步骤分离髓磷脂膜，这些步骤包括密度梯度离心和差速离心。在等渗蔗糖中对神经组织进行匀浆会使髓鞘从轴突上剥离并形成相对较大的髓磷脂囊泡。髓磷脂囊泡的大尺寸，以及髓磷脂膜的密度低于其他生物膜这一事实，使得差速离心和密度梯度离心成为分离这种膜的主要工具。本单元概述了三种方案：从中枢神经系统（CNS）中分离高度纯化的髓磷脂组分；将高度纯化的CNS髓磷脂组分分离成不同密度的亚组分；以及从周围神经系统（PNS）中分离髓磷脂。
