Lmo genes regulate behavioral responses to ethanol in Drosophila melanogaster and the mouse.
- Authors
- Lasek, Amy W; Giorgetti, Francesco; Berger, Karen H; Tayor, Stacy; Heberlein, Ulrike
- Year
- 2011
- Journal
- Alcoholism, clinical and experimental research
- PMID
- 21599714
- DOI
- 10.1111/j.1530-0277.2011.01506.x
- PMCID
- PMC3166402
BACKGROUND: Previous work from our laboratory demonstrated a role for the Drosophila Lim-only (dLmo) gene in regulating behavioral responses to cocaine. Herein, we examined whether dLmo influences the flies' sensitivity to ethanol's sedating effects. We also investigated whether 1 of the mammalian homologs of dLmo, Lmo3, is involved in behavioral responses to ethanol in mice. METHODS: To examine dLmo function in ethanol-induced sedation, mutant flies with reduced or increased dLmo expression were tested using the loss of righting (LOR) assay. To determine whether mouse Lmo3 regulates behavioral responses to ethanol, we generated transgenic mice expressing a short-hairpin RNA targeting Lmo3 for RNA interference-mediated knockdown by lentiviral infection of single cell embryos. Adult founder mice, expressing varying amounts of Lmo3 in the brain, were tested using ethanol loss-of-righting-reflex (LORR) and 2-bottle choice ethanol consumption assays. RESULTS: We found that in flies, reduced dLmo activity increased sensitivity to ethanol-induced sedation, whereas increased expression of dLmo led to increased resistance to ethanol-induced sedation. In mice, reduced levels of Lmo3 were correlated with increased sedation time in the LORR test and decreased ethanol consumption in the 2-bottle choice protocol. CONCLUSIONS: These data describe a novel and conserved role for Lmo genes in flies and mice in behavioral responses to ethanol. These studies also demonstrate the feasibility of rapidly translating findings from invertebrate systems to mammalian models of alcohol abuse by combining RNA interference in transgenic mice and behavioral testing.
dLmo/Bx mutants display altered resistance to ethanol sedation and no change in ethanol absorption. (A) Ethanol sedation curves indicating that EP1306 flies show increased sensitivity and BxJ flies show decreased sensitivity to ethanol sedation in the loss of righting (LOR) assay when compared with control (Ctl) flies. Ctl flies were in the w;iso genetic background. (B) The median sedation time (ST50)—the time required for half of the ethanol-exposed flies to show LOR—was calculated by linear interpolation. Error bars represent SEM, and asterisks denote statistical significance by one-way ANOVA followed by post hoc Holm-Sidak testing (**p < 0.01; ***p < 0.001; n = 9 to 12, where n is the number of samples, not the number of flies). (C) Samples of mutant and control flies were exposed to ethanol vapor under identical conditions as for behavioral assays, and were snap-frozen after 15 minutes and processed for internal ethanol absorption. No significant difference was seen between mutant and control flies (n = 10 or 11).
Efficacy of short-hairpin RNAs (shRNAs) targeting Lmo3 for RNA interference in Neuro-2a cells. (A) Schematic illustrating the position of shRNAs targeting the Lmo3 transcript. Colored boxes show shRNA location, open box illustrates the protein coding region, and numbers indicate the nucleotide position along the Lmo3 mRNA. (B) Expression of Lmo3 in Neuro-2a cells transfected with lentiviral plasmids expressing shRNAs targeting Lmo3 (shLmo3.5, shLmo3.7, and shLmo3.8) or the control shScr. RNA was isolated 2 days after transfection and subjected to qPCR. Lmo3 expression is normalized to expression of the housekeeping gene, Gapdh. Asterisk indicates a significant difference between the shLmo3 constructs and shScr by one-way ANOVA (*p = 0.007; n = 3). (C, D) Lmo4 (C) and Lmo1 (D) expression in transfected Neuro-2a samples described in (B).
Characterization of transgenic mice expressing shLmo3.8 or shScr. (A–F) Green fluorescent protein (GFP) fluorescence in representative sagittal adult brain sections of transgenic mice infected at the single-cell embryo stage with lentivirus encoding shLmo3.8 (B, D, F) or shScr (A, C, E). GFP expression from the viral vector is visible in cell bodies (bright spots) and processes of neurons throughout the brain, including the cortex (A, B, Ctx), nucleus accumbens (C, D, Acb), and cerebellum (E, F, Cer). Hazy green areas represent background autofluorescence from the tissue, rather than infection per se. (G) Expression of GFP and Lmo3 transcript levels were negatively correlated in the forebrains of transgenic shLmo3.8 (closed circles, n = 12), but not shScr (open circles, n = 8) mice. GFP (x-axis) and Lmo3 (y-axis) expression values were normalized relative to expression of the housekeeping gene Gapdh.
Correlations between Lmo3 expression and behavioral responses to ethanol in transgenic shLmo3.8 and shScr mice. In all graphs, normalized Lmo3 expression is plotted on the x-axis and the behavioral response on the y-axis. Mice expressing shLmo3.8 (n = 12) are represented by closed circles and shScr (n = 8) by open circles. (A) Negative correlation between Lmo3 expression and ethanol sedation in the loss-of-righting-reflex test with a 3.2 g/kg dose of ethanol. (B, C) Positive correlation between Lmo3 expression and 2-bottle choice ethanol consumption of 6% (B) and 10% (C) ethanol. (D) Water consumption and Lmo3 expression were not significantly correlated.
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| An inexpensive, scalable behavioral assay for measuring ethanol sedation sensitivity and rapid tolerance in Drosophila. | Sandhu S et al. | — | 2015 | → |
| Drosophila and Caenorhabditis elegans as Discovery Platforms for Genes Involved in Human Alcohol Use Disorder. | Grotewiel M et al. | — | 2015 | → |
| Peer Influence, Genetic Propensity, and Binge Drinking: A Natural Experiment and a Replication. | Guo G et al. | — | 2015 | → |
| Beadex function in the motor neurons is essential for female reproduction in Drosophila melanogaster. | Kairamkonda S et al. | — | 2014 | → |
| Ethanol and cocaine: environmental place conditioning, stereotypy, and synergism in planarians. | Tallarida CS et al. | — | 2014 | → |
| Genetics and genomics of alcohol sensitivity. | Morozova TV et al. | — | 2014 | → |
| Increased behavioral responses to ethanol in Lmo3 knockout mice. | Savarese A et al. | — | 2014 | → |
| The buzz on caffeine in invertebrates: effects on behavior and molecular mechanisms. | Mustard JA | — | 2014 | → |
| The FlyBar: administering alcohol to flies. | van der Linde K et al. | — | 2014 | → |
| A meta-analysis of two genome-wide association studies to identify novel loci for maximum number of alcoholic drinks. | Kapoor M et al. | — | 2013 | → |
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