Effects of ethanol on neuroblastoma cells in culture: role of gangliosides in neuritogenesis and substrate adhesion.
Murine Neuro-2A neuroblastoma cells were exposed to ethanol in culture under two experimental paradigms: (1) short-term (24 hr or less) and low concentrations (0.05 to 0.5%; 8.5 to 86 mM) and (2) long-term (48 hr at 0.5%; 86 mM). Long-term ethanol exposure did not affect Neuro-2A viability, determined by DNA synthesis or the ability to exclude Trypan Blue. Similarly, long-term ethanol treatment did not inhibit differentiation, exhibited by the extension of neurites, promoted by either dibutyryl-cyclic-AMP or by incubation with exogenous ganglioside GM1. The incorporation of exogenous ganglioside GM1 into plasma membranes was not influenced by varying concentrations of ethanol (up to 1.2%; 204 mM). In contrast, ethanol did influence Neuro-2A cell attachment to collagen in a dualistic manner. During short-term ethanol exposure, cell attachment was enhanced. However, when cells were initially exposed to ethanol for 48 hr a marked inhibition of subsequent attachment was observed. Long-term ethanol exposure also inhibited attachment to other substrata, including laminin, fibronectin and vitronectin. Incubation of Neuro-2A cells with either exogenous ganglioside GM1 or a mixture of brain gangliosides partially reversed the inhibition of attachment to collagen. This reversal did not appear to be due to any one particular ganglioside structure, however. Mixed brain gangliosides were fractionated into three fractions, according to the number of sialic acid residues. Each of the three fractions were equally effective in partially restoring Neuro-2A cell attachment to collagen after long-term ethanol treatment. The results suggest that the mechanism by which these effects occur is at the level of plasma membrane fluidity, because both ethanol and glycosphingolipid content are known to influence membrane lateral mobility, although other mechanisms, such as changes in headgroup hydration, are possible.
Leskawa
K C
KC
Department of Anatomical Sciences and Neurobiology, School of Medicine, University of Louisville, Kentucky 40292, USA.
Maddox
T
T
Webster
K A
KA
D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism*; D002417:Cattle; D002448:Cell Adhesion; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D000431:Ethanol / Q000494:pharmacology*; D005677:G(M1) Ganglioside / Q000502:physiology; D005732:Gangliosides / Q000502:physiology*; D051379:Mice; D016501:Neurites / Q000187:drug effects* / Q000648:ultrastructure; D009447:Neuroblastoma / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D002492:Central Nervous System Depressants; D005732:Gangliosides; D005677:G(M1) Ganglioside; D000431:Ethanol