Differential regulation of alcohol consumption and reward by the transcriptional cofactor LMO4

Abstract

Repeated alcohol exposure leads to changes in gene expression that are thought to underlie the transition from moderate to excessive drinking. However, the mechanisms by which these changes are integrated into a maladaptive response that leads to alcohol dependence are not well understood. One mechanism could involve the recruitment of transcriptional co-regulators that bind and modulate the activity of transcription factors. Our results indicate that the transcriptional regulator LMO4 is one such candidate regulator. Lmo4-deficient mice (Lmo4gt/+) consumed significantly more and showed enhanced preference for alcohol in a 24 h intermittent access drinking procedure. shRNA-mediated knockdown of Lmo4 in the nucleus accumbens enhanced alcohol consumption, whereas knockdown in the basolateral amygdala (BLA) decreased alcohol consumption and reduced conditioned place preference for alcohol. To ascertain the molecular mechanisms that underlie these contrasting phenotypes, we carried out unbiased transcriptome profiling of these two brain regions in wild type and Lmo4gt/+ mice. Our results revealed that the transcriptional targets of LMO4 are vastly different between the two brain regions, which may explain the divergent phenotypes observed upon Lmo4 knockdown. Bioinformatic analyses revealed that Oprk1 and genes related to the extracellular matrix (ECM) are important transcriptional targets of LMO4 in the BLA. Chromatin immunoprecipitation revealed that LMO4 bound Oprk1 promoter elements. Consistent with these results, disruption of the ECM or infusion of norbinaltorphimine, a selective kappa opioid receptor antagonist, in the BLA reduced alcohol consumption. Hence our results indicate that an LMO4-regulated transcriptional network regulates alcohol consumption in the BLA.

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Fig. 1: Lmo4 haploinsuffficiency increase alcohol consumption and preference.
Fig. 2: Knockdown of LMO4 in the NAc increases alcohol consumption.
Fig. 3: Knockdown of LMO4 in the BLA reduces alcohol consumption and conditioned place preference.
Fig. 4: Transcriptome analysis identifies ECM-related genes as transcriptional targets of LMO4 that regulate alcohol consumption.
Fig. 5: BLA KORs promote alcohol consumption.

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Acknowledgements

This work was supported by NIH grants AA025244 (ROM) and AA027293 (RM), and by Graduate Research Fellowship DGE-1110007 from the National Science Foundation to MBP. We thank Haridha Shivram for help with ChIP and Heather Aziz for help with Gas Chromatography.

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RM, MBP, and ROM designed the experiments. RM and AB performed stereotaxic surgeries. RM, AB, TT, and MTP performed behavioral experiments and analyzed the data. MBP performed in situ hybridization experiments. GRT, RM, and RDM did bioinformatics analysis of RNAseq data. RM, MBP, and ROM wrote the paper.

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Correspondence to Rajani Maiya.

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Maiya, R., Pomrenze, M.B., Tran, T. et al. Differential regulation of alcohol consumption and reward by the transcriptional cofactor LMO4. Mol Psychiatry (2020). https://doi.org/10.1038/s41380-020-0706-8

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