Correlation between morphological and biochemical effects of ethanol on neuroblast-enriched cultures derived from three-day-old chick embryos.
We have shown that ethanol exposure during embryogenesis affects a variety of parameters of neuronal growth. In this study we examined the direct effects of ethanol exposure on developing neuroblasts in culture. Neuroblast-enriched cultures derived from 3-day-old whole chick embryos were grown in the presence of ethanol at doses ranging from 12.5 to 50 mM from culture day 3-14. Cholinergic and GABAergic phenotypic expression were both significantly reduced following ethanol exposure as assessed by the activities of choline acetyltransferase and glutamate decarboxylase, respectively. Morphometric analysis of the growth patterns showed significant differences between control and ethanol-treated cultures. Control cultures exhibited the characteristic pattern of growth consisting of neuronal aggregation with neuritic arborization, i.e., neuritic bundles and fasciculation. Cultures grown in ethanol from culture day 3 consisted of aggregates that measured significantly greater in size than those observed in control cultures. In addition, in ethanol-treated cultures, the primary pattern of neuritic bundles was replaced by a complex network of individual neurites radiating from the central aggregate, forming a defined "neuritic field." Morphometric analysis revealed that both neurite number and neurite length were significantly reduced in ethanol-treated cultures. The biochemical data confirm earlier reports from this laboratory suggesting that ethanol exposure during early embryogenesis alters the normal neuronal pattern of phenotypic expression. In addition, we have presented evidence in this study that ethanol alters the morphological growth patterns of developing neurons. Although ethanol does not alter the ability of these cells to aggregate, there is a significant alteration in neuritic outgrowth.(ABSTRACT TRUNCATED AT 250 WORDS)
Kentroti
S
S
Department of Pharmacology, University of Colorado School of Medicine, Denver 80262.
Vernadakis
A
A
D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000494:pharmacology*; D005968:Glutamate Decarboxylase / Q000378:metabolism*; D007700:Kinetics; D009419:Nerve Tissue Proteins / Q000032:analysis; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*
D009419:Nerve Tissue Proteins; D000431:Ethanol; D002795:Choline O-Acetyltransferase; D005968:Glutamate Decarboxylase