Document Type
Article
Original Publication Date
2014
Journal/Book/Conference Title
Alcoholism: Clinical and Experimental Research
Volume
38
Issue
6
First Page
1582
Last Page
1593
DOI of Original Publication
10.1111/acer.12421
Date of Submission
July 2015
Abstract
Background
The fruit fly Drosophila melanogaster has been used extensively to investigate genetic mechanisms of ethanol-related behaviors. Many past studies in flies, including studies from our laboratory, have manipulated gene expression using transposons carrying the genetic-phenotypic marker mini-white, a derivative of the endogenous gene white. Whether the mini-white transgenic marker or the endogenous white gene influence behavioral responses to acute ethanol exposure in flies has not been systematically investigated.
Methods
We manipulated mini-white and white expression via (i) transposons marked with mini-white, (ii) RNAi against mini-white and white and (iii) a null allele of white. We assessed ethanol sensitivity and tolerance using a previously described eRING assay (based on climbing in the presence of ethanol) and an assay based on ethanol-induced sedation.
Results
In eRING assays, ethanol-induced impairment of climbing correlated inversely with expression of the mini-white marker from a series of transposon insertions. Additionally, flies harboring a null allele of white or flies with RNAi-mediated knockdown of mini-white were significantly more sensitive to ethanol in eRING assays than controls expressing endogenous white or the mini-white marker. In contrast, ethanol sensitivity and rapid tolerance measured in the ethanol sedation assay were not affected by decreased expression of mini-white or endogenous white in flies.
Conclusions
Ethanol sensitivity measured in the eRING assay is noticeably influenced by white and mini-white, making eRING problematic for studies on ethanol-related behavior in Drosophila using transgenes marked with mini-white. In contrast, the ethanol sedation assay described here is a suitable behavioral paradigm for studies on ethanol sedation and rapid tolerance in Drosophila including those that use widely available transgenes marked with mini-white.
Rights
Copyright © 2014 by the Research Society on Alcoholism. This is the peer reviewed version of the following article: Chan, R. F., Lewellyn, L., DeLoyht, J. M., Sennett, K., Coffman, S., Hewitt, M., Bettinger, J. C., Warrick, J. M. and Grotewiel, M. (2014), Contrasting Influences of Drosophila white/mini-white on Ethanol Sensitivity in Two Different Behavioral Assays. Alcoholism: Clinical and Experimental Research, 38: 1582–1593, which has been published in final form at http://dx.doi.org/10.1111%2Facer.12421. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Is Part Of
VCU Human and Molecular Genetics Publications
Comments
Published in final edited form as: Alcohol Clin Exp Res. 2014 Jun; 38(6): 1582–1593. doi: 10.1111/acer.12421. PMCID: PMC4049357 NIHMSID: NIHMS577293