Myelination in cerebellar slice cultures: development of a system amenable to biochemical analysis.
Myelin deposition and maintenance are critical to proper function of the mammalian nervous system. Previous investigations of myelination in the central nervous system (CNS) were hampered by the lack of an in vitro system that can faithfully reproduce in vivo events yet is amenable to biochemical investigation. We have developed a procedure, based on organotypic cultures, which permits efficient preparation of large numbers of cerebellar slice cultures that can be easily manipulated. Cultures have been examined to document myelination biochemically (by incorporation of [35S]sulfate into sulfolipids), immunohistochemically (by labeling the myelin components myelin basic protein and galactocerebroside), and morphologically (by both light and electron microscopy). We tested the effects of biologically active peptides and antibodies on myelination in the thin slices. The results indicate that the cultures provide an in vitro system that can be used to examine specific cellular events that occur during CNS myelination.
Notterpek
L M
LM
Department of Biological Chemistry, UCLA School of Medicine 90024-1737.
Bullock
P N
PN
Malek-Hedayat
S
S
Fisher
R
R
Rome
L H
LH
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D046508:Culture Techniques; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D004676:Myelin Basic Protein / Q000276:immunology / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism / Q000502:physiology*; D009412:Nerve Fibers / Q000502:physiology; D016900:Neurofilament Proteins / Q000276:immunology / Q000378:metabolism; D009457:Neuroglia / Q000648:ultrastructure; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D013462:Sulfur Radioisotopes
D015415:Biomarkers; D004676:Myelin Basic Protein; D016900:Neurofilament Proteins; D013462:Sulfur Radioisotopes