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Isolation of myelin
Jorge N Larocca1, Williams T Norton
1Albert Einstein College of Medicine, Yeshiva University, Bronx, New York, USA.
Abstract:
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).
Insights
This study details methods for isolating myelin, a key component of nervous tissue. Optimized protocols enable purification of myelin membranes from both central and peripheral nervous systems.
Area of Science:
- Neuroscience
- Biochemistry
- Cell Biology
Background:
- Myelin is a crucial lipid-rich insulating sheath surrounding nerve axons.
- Effective isolation of myelin membranes is essential for studying its composition and function.
Purpose of the Study:
- To outline established and reliable protocols for myelin preparation.
- To enable the isolation and subfractionation of myelin from nervous tissue.
Main Methods:
- Homogenization of nervous tissue in isotonic sucrose solution.
- Differential centrifugation and density gradient centrifugation for myelin vesicle isolation.
- Specific protocols for central nervous system (CNS) and peripheral nervous system (PNS) myelin.
Main Results:
- Myelin sheath peels from axons, forming large, low-density vesicles.
- Density gradient and differential centrifugation effectively isolate these vesicles.
- Protocols yield highly-purified CNS myelin and allow for subfractionation by density.
Conclusions:
- Established centrifugation techniques are effective for myelin isolation.
- The described methods facilitate the study of myelin structure and biochemistry.
- These protocols are applicable to both CNS and PNS myelin research.
