GM1 enhances the association of neuron-specific MAP2 with actin in MAP2-transfected 3T3 cells.
The ganglioside GM1 is a glycosphingolipid which enhances process formation of several neuronal lines and potentiates some growth factor-mediated responses. Previously we have shown that 24 h exposure of Neuro 2a cells to GM1 mobilized the neuron-specific microtubule-associated protein, MAP2, away from microtubule-rich areas to areas of neurite sprouting where MAP2 was more closely associated with the subcortical actin network. To examine the role of GM1 in fostering the shift of the association of MAP2 from tubulin to actin, NIH 3T3 cells were co-transfected with pHook-1, which expresses a surface antigen, and a construct expressing MAP2. Transfected cells were selected with magnetic beads coated with a hapten that binds to the expressed surface antigen and treated with 150 microg/ml GM1 for 18-24 h. Actin and MAP2 or tubulin and MAP2 were immunolocalized and examined with confocal microscopy. MAP2 was found throughout the cytoplasm as well as associated with actin filaments. As observed previously with Neuro 2a, GM1 treatment of transfected fibroblasts redistributed the MAP2 away from direct association with microtubules to peripheral areas where the association of MAP2 with actin was enhanced. GM1 did not induce neurite-like processes in MAP2-transfected cells. Treatment with cytochalasin B, which is reported to result in process formation, also did not induce neurite-like processes. These studies suggest that GM1's ability to mobilize MAP2 and promote its association with actin is not restricted to neurons.
Colella
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R
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, 500 S. Preston Street, Louisville, KY 40292, USA. r0cole01@gwise.louisville.edu
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Hodges
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Wilkey
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Roisen
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FJ
D016475:3T3 Cells; D000199:Actins / Q000378:metabolism*; D000818:Animals; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D005677:G(M1) Ganglioside / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D005677:G(M1) Ganglioside