Distinct stages of myelination regulated by gamma-secretase and astrocytes in a rapidly myelinating CNS coculture system.
Mechanistic studies of CNS myelination have been hindered by the lack of a rapidly myelinating culture system. Here, we describe a versatile CNS coculture method that allows time-lapse microscopy and molecular analysis of distinct stages of myelination. Employing a culture architecture of reaggregated neurons fosters extension of dense beds of axons from purified retinal ganglion cells. Seeding of oligodendrocyte precursor cells on these axons results in differentiation and ensheathment in as few as 3 days, with generation of compact myelin within 6 days. This technique enabled (1) the demonstration that oligodendrocytes initiate new myelin segments only during a brief window early in their differentiation, (2) identification of a contribution of astrocytes to the rate of myelin wrapping, and (3) molecular dissection of the role of oligodendrocyte gamma-secretase activity in controlling the ensheathment of axons. These insights illustrate the value of this defined system for investigating multiple aspects of CNS myelination.
Watkins
Trent A
TA
Stanford University School of Medicine, Department of Neurobiology, Fairchild Science Building D235, Stanford, CA 94305-5125, USA. trentw@gene.com
Emery
Ben
B
Mulinyawe
Sara
S
Barres
Ben A
BA
D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism*; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D002450:Cell Communication / Q000502:physiology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology / Q000378:metabolism*; D018920:Coculture Techniques / Q000379:methods; D020439:Growth Cones / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D009186:Myelin Sheath / Q000378:metabolism; D009413:Nerve Fibers, Myelinated / Q000378:metabolism*; D055495:Neurogenesis / Q000502:physiology*; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D013997:Time Factors
D053829:Amyloid Precursor Protein Secretases