Chronic ethanol exposure increases microtubule content in PC12 cells.
BACKGROUND
Chronic ethanol exposure has been shown to result in changes in neuronal cyto-architecture such as aberrant sprouting and alteration of neurite outgrowth. In PC12 cells, chronic ethanol treatment produces an increase in Nerve Growth Factor (NGF)-induced neurite outgrowth that appears to require the epsilon, but not delta, isoform of Protein Kinase C (PKC). Neurites contain a core of microtubules that are formed from polymerization of free-tubulin. Therefore, it would be expected that an increase in neurite outgrowth would correlate with an increase in microtubule content. We examined the effect of chronic ethanol exposure on microtubule content in PC12 cells and the role of PKC epsilon and delta in ethanol's effect on microtubule levels.


RESULTS
Chronic ethanol exposure of wild-type and vector control PC12 cells resulted in a significant increase in microtubule content and a corresponding decrease in free tubulin. There was also a significant increase in microtubule content in PC12 cells expressing a dominate-negative inhibitor of epsilon PKC; cells which have previously been shown to have no ethanol-induced increase in neurite outgrowth. In contrast, ethanol had no effect on microtubule content in PC12 cells expressing a dominate-negative inhibitor of delta PKC.


CONCLUSIONS
These results suggest that chronic ethanol exposure alters the relative ratio of free tubulin to microtubule-associated tubulin, an important component of the cytoskeleton. Further, the data from the PKC dominant-negative cell lines suggest that the effects of ethanol on microtubule content do not correlate with the effects of ethanol on neurite outgrowth. The delta isoform of PKC appears to be necessary for the ethanol-induced increase in microtubule content. These studies demonstrate an effect of chronic ethanol exposure which may contribute to previously documented alterations of neuronal cyto-architecture.
Reiter-Funk
Cindy K
CK
Dept. of Human Anatomy and Medical Neurobiology, College of Medicine, Texas A&M University System Health Science Center, College Station, TX 77843, USA. creiter@scripps.edu <creiter@scripps.edu>
Dohrman
Douglas P
DP
D000818:Animals; D000431:Ethanol / Q000008:administration & dosage*; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats; D014404:Tubulin / Q000187:drug effects / Q000378:metabolism
D014404:Tubulin; D000431:Ethanol