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An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila
Published on: April 15, 2015
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A DNA element in the slo gene modulates ethanol tolerance
Harish R Krishnan1, Xiaolei Li1, Alfredo Ghezzi1
1The Waggoner Center for Alcohol and Addiction Research, Department of Neuroscience, The University of Texas at Austin, Austin, TX, USA.
Alcohol (Fayetteville, N.Y.)
|March 20, 2016
Summary
Researchers identified a DNA element, called 6b, within the slo gene that regulates functional ethanol tolerance in Drosophila. Removing this element significantly extended the duration of tolerance after ethanol sedation.
Area of Science:
- Neuroscience
- Genetics
- Pharmacology
Background:
- The slo gene in Drosophila encodes BK-type Ca(2+)-activated K(+) channels, crucial for rapid functional tolerance to ethanol sedation.
- Drosophila serve as an ideal model for studying functional ethanol tolerance due to their lack of metabolic tolerance acquisition.
- Previous studies indicate slo gene mutations impair tolerance acquisition, and ethanol sedation upregulates slo expression in the nervous system.
Purpose of the Study:
- To map a DNA regulatory element in the slo gene's transcriptional control region using ethanol-induced histone acetylation.
- To functionally test the identified DNA element's role in producing functional ethanol tolerance.
- To investigate the impact of a specific DNA element (6b) on the duration of ethanol tolerance.
Main Methods:
- Utilized chromatin immunoprecipitation (ChIP) assays to map histone acetylation changes in response to ethanol sedation.
- Identified an ethanol-responsive DNA element (6b) located between neural promoters of the slo gene.
- Assessed functional ethanol tolerance by removing the 6b element from the endogenous slo gene and using an ethanol-vapor recovery assay.
Main Results:
- Ethanol sedation increased histone acetylation over the 6b DNA element, indicating its involvement in the ethanol response.
- Removal of the 6b element from the slo gene significantly extended the duration of functional ethanol tolerance.
- Functional ethanol tolerance persisted for over 21 days after sedation in flies lacking the 6b element, compared to ~10 days in controls.
Conclusions:
- Mapping ethanol-induced histone acetylation is an effective strategy for identifying gene regulatory elements involved in ethanol response.
- The identified 6b DNA element plays a critical role in mediating the duration of functional ethanol tolerance in Drosophila.
- This conserved DNA element is important for a behaviorally relevant ethanol response, highlighting its significance in neuroadaptation.
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