The effects of cytoskeletal altering agents on the surface topography of GM1 in neuro-2A neuroblastoma cell membranes.
Neuro-2a murine neuroblastomal cells exposed to exogenous ganglioside undergo increased neuritogenesis in vitro. To determine if the distribution of exogenous ganglioside (GM1) in neuronal membranes is related to neuritogenesis, the surface topography of exogenous ganglioside in these cells was examined by localization with cholera toxin B-FITC. Following exposure to exogenous ganglioside, levels of fluorescent label appeared similar on perikaryal and neuritic surfaces. Scanning electron microscopic studies using protein G-gold to label antibody against exogenous ganglioside confirmed these observations at higher magnification. Within the general labelling pattern, occasionally labelled material was observed which seemed to form short linear arrays. This suggested that elements of the cytoskeleton might be influencing the surface distribution of exogenous ganglioside. To examine this possibility, Neuro-2a cells were exposed to agents known to alter the stability of specific cytoskeletal components, after which the general distribution of exogenous ganglioside was determined. Treatment with Colcemid, which disrupted microtubules, resulted in restriction of most exogenous ganglioside-positive label to the perikaryal surfaces. In contrast, exposure to taxol which enhanced microtubule stability diminished perikaryal fluorescence and increased neuritic labelling. The disruption of cytochalasin D-sensitive microfilaments did not influence the topographic distribution of exogenous ganglioside. Under the experimental conditions employed, mean neuritic lengths for Colcemid- and taxol-treated cells were nearly equal, indicating that altered neuritic length resulting from treatment with cytoskeletal agents was not a major factor in the redistribution of exogenous ganglioside. These studies suggest that microtubules play a role in determining the distribution of recently incorporated ganglioside in neuronal plasma membranes.
Fentie
I H
IH
Department of Anatomical Sciences and Neurobiology, School of Medicine, University of Louisville, KY 40292.
Roisen
F J
FJ
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure*; D002772:Cholera Toxin; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects*; D003703:Demecolcine / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D005677:G(M1) Ganglioside / Q000032:analysis; D005732:Gangliosides / Q000494:pharmacology*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008870:Microtubules / Q000187:drug effects; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D009447:Neuroblastoma / Q000648:ultrastructure*; D017239:Paclitaxel / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D005456:Fluorescent Dyes; D005732:Gangliosides; D015638:Cytochalasin D; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D016650:Fluorescein-5-isothiocyanate; D017239:Paclitaxel; D003703:Demecolcine