title: Anti-beta 1 integrin antibody inhibits Schwann cell myelination.
abstract: Schwann cells (SCs) co-cultured with sensory neurons require ascorbate supplementation for basal lamina assembly and differentiation into myelinating cells. The ascorbate requirement can be bypassed by adding a purified basal lamina component, laminin, to SC/neuron co-cultures. We have examined the role of laminin receptors, namely, the beta 1 subfamily of integrins, in the process of myelination. We demonstrate by immunostaining or immunoprecipitation that undifferentiated SCs in contact with axons express large amounts of the beta 1 subunit in association with the alpha 1 or alpha 6 subunit. In co-cultures of myelinating SCs, alpha 1 beta 1 is no longer present, alpha 6 beta 1 is still present but at reduced levels, and alpha 6 beta 4 is expressed at much higher levels than in co-cultures of undifferentiated SCs. Immunogold labelling at the electron microscope level suggested that beta 1 integrins are randomly distributed on undifferentiated SCs, become localized to the SC surface contacting basal lamina in differentiating SCs before the onset of myelination, and are not detected on myelinating SCs. Fab fragments of beta 1 function-blocking antibody block both attachment of isolated SCs to laminin and formation of myelin sheaths by SCs co-cultured with neurons in ascorbate-supplemented medium. SCs unable to myelinate in the presence of the anti-beta 1 antibody assemble patchy basal lamina that is only loosely attached to the cell surface and in some cases appears to be detaching from the membrane. In contrast, an alpha 1 beta 1 function-blocking antibody only partially blocks attachment of isolated SCs to laminin but has no inhibitory effect on SC myelination. These results are consistent with the hypothesis that a member of the beta 1 subfamily of integrins other than alpha 1 beta 1 binds laminin present in basal lamina to the SC surface and transduces signals that are critical for initiation of SC differentiation into a myelinating cell.
authors.affiliation: Chambers Family Electron Microscopy Laboratory, University of Miami School of Medicine, Florida 33136.
authors.forename: C
authors.initials: C
authors.lastname: Fernandez-Valle
authors.forename: L
authors.initials: L
authors.lastname: Gwynn
authors.forename: P M
authors.initials: PM
authors.lastname: Wood
authors.forename: S
authors.initials: S
authors.lastname: Carbonetto
authors.forename: M B
authors.initials: MB
authors.lastname: Bunge
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology; D002448:Cell Adhesion; D002454:Cell Differentiation; D007158:Immunologic Techniques; D039222:Integrin alpha1beta1; D039121:Integrin alpha6beta1; D019012:Integrin beta1; D016023:Integrins / Q000378:metabolism / Q000502:physiology*; D007425:Intracellular Membranes / Q000378:metabolism; D007797:Laminin; D009186:Myelin Sheath / Q000502:physiology*; D011233:Precipitin Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000166:cytology / Q000502:physiology*
chemical_list: D039222:Integrin alpha1beta1; D039121:Integrin alpha6beta1; D019012:Integrin beta1; D016023:Integrins; D007797:Laminin