title: Multicolor analysis of oligodendrocyte morphology, interactions, and development with Brainbow.
abstract: Oligodendrocytes are the myelinating cells of the central nervous system. Multiple markers are available to analyze the populations of oligodendroglial cells and their precursors during development and in pathological conditions. However, the behavior of oligodendrocytes remains poorly characterized in vivo, especially at the level of individual cells. Studying this aspect has been impaired so far by the lack of suitable methods for visualizing single oligodendrocytes, their processes, and their interactions during myelination. Here, we have used multicolor labeling technology to single-out simultaneously many individual oligodendrocytes in the postnatal mouse optic nerve. This method is based on Brainbow, a transgenic system for stochastic expression of multiple fluorescent protein genes through Cre-lox recombination, previously used for visualizing axons and neurons. We used tamoxifen-inducible recombination in myelinating cells of Brainbow transgenic mice to obtain multicolor labeling of oligodendrocytes. We show that the palette of colors expressed by labeled oligodendrocytes is tamoxifen dependent, with the highest doses producing the densest and most colorful labeling. At low doses of tamoxifen, the morphology of single or small clusters of fluorescent oligodendrocytes can be studied during postnatal development and in adult. Internodes are labeled to their extremities, revealing nodes of Ranvier. The new mouse model presented here opens new possibilities to explore the organization and development of the oligodendrocyte network with single-cell resolution.
authors.affiliation: INSERM, UMRS_U968, Institut de la Vision, Paris, F-75012, France; Sorbonne Universités, UPMC Univ Paris 06, Institut de la Vision, Paris, F-75012, France; CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Dumas
authors.forename: Céline
authors.initials: C
authors.lastname: Heitz-Marchaland
authors.forename: Stephane
authors.initials: S
authors.lastname: Fouquet
authors.forename: Ueli
authors.initials: U
authors.lastname: Suter
authors.forename: Jean
authors.initials: J
authors.lastname: Livet
authors.forename: Caroline
authors.initials: C
authors.lastname: Moreau-Fauvarque
authors.forename: Alain
authors.initials: A
authors.lastname: Chédotal
mesh_terms: D000818:Animals; D005455:Fluorescent Antibody Technique / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009413:Nerve Fibers, Myelinated / Q000378:metabolism*; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism; D009900:Optic Nerve / Q000166:cytology*; D011995:Recombination, Genetic; D013194:Staining and Labeling / Q000379:methods*; D013269:Stochastic Processes; D013629:Tamoxifen / Q000008:administration & dosage; D019076:Transgenes
chemical_list: D008164:Luminescent Proteins; D013629:Tamoxifen
keywords: imaging; myelination; transgenic mouse