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Cell-type and sex-specific rhythmic gene expression in the nucleus accumbens

Molecular Psychiatry (2024)Cite this article

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Abstract

Circadian rhythms are critical for human health and are highly conserved across species. Disruptions in these rhythms contribute to many diseases, including psychiatric disorders. Previous results suggest that circadian genes modulate behavior through specific cell types in the nucleus accumbens (NAc), particularly dopamine D1-expressing medium spiny neurons (MSNs). However, diurnal rhythms in transcript expression have not been investigated in NAc MSNs. In this study we identified and characterized rhythmic transcripts in D1- and D2-expressing neurons and compared rhythmicity results to homogenate as well as astrocyte samples taken from the NAc of male and female mice. We find that all cell types have transcripts with diurnal rhythms and that top rhythmic transcripts are largely core clock genes, which peak at approximately the same time of day in each cell type and sex. While clock-controlled rhythmic transcripts are enriched for protein regulation pathways across cell type, cell signaling and signal transduction related processes are most commonly enriched in MSNs. In contrast to core clock genes, these clock-controlled rhythmic transcripts tend to reach their peak in expression about 2-h later in females than males, suggesting diurnal rhythms in reward may be delayed in females. We also find sex differences in pathway enrichment for rhythmic transcripts peaking at different times of day. Protein folding and immune responses are enriched in transcripts that peak in the dark phase, while metabolic processes are primarily enriched in transcripts that peak in the light phase. Importantly, we also find that several classic markers used to categorize MSNs are rhythmic in the NAc. This is critical since the use of rhythmic markers could lead to over- or under-enrichment of targeted cell types depending on the time at which they are sampled. This study greatly expands our knowledge of how individual cell types contribute to rhythms in the NAc.

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Fig. 1: Rhythmicity in the NAc varies by cell type.
Fig. 2: Rhythmic transcripts vary across cell type and sex in the NAc.
Fig. 3: Rhythmic transcripts tend to peak later in females than males.
Fig. 4: Rhythmic transcripts peaking in the light phase contribute to largely distinct enriched pathways.
Fig. 5: Rhythmicity should be considered when cell-type-specific markers are selected.
Fig. 6: Model summarizing enriched rhythmic processes in MSNs across time of day and sex.

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Data availability

Sequencing data are available in the Gene Expression Omnibus (GEO) database. The code used for analysis is available at GitHub.

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Acknowledgements

We would like to thank Mariah Hildebrand and Ioannis Migias for animal care and genotyping. We thank Dr. Mary Kay Lobo for providing the Ribotag transgenic mice, as well as for technical assistance with co-immunoprecipitations. This project used the University of Pittsburgh Health Sciences Sequencing Core at UPMC Children’s Hospital of Pittsburgh (RNA sequencing) and the University of Pittsburgh Center for Research Computing, RRID:SCR_022735, through the resources provided. Specifically, this work used the HTC cluster, which is supported by NIH award number S10OD028483. This work was funded by the National Institutes of Health: DA039865, DA046346, MH111601, MH106460 (PI: Colleen McClung, PhD), DA046117, DA055064, L60DA054665 (PI: LMD) and MH128763 (PI: KDK) as well as the Brain and Behavior Research Foundation (29386, LMD and 30823 (P&S Fund), Ketchesin).

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Authors and Affiliations

  1. Department of Psychiatry, Translational Neuroscience Program, University of Pittsburgh School of Medicine, 15219, Pittsburgh, PA, USA

    Lauren M. DePoy, Kaitlyn A. Petersen, Kyle D. Ketchesin, Ross C. Matthaei, Megan S. Perez, Darius Becker-Krail, Madeline R. Scott & Colleen A. McClung

  2. Center for Neuroscience, University of Pittsburgh, 15261, Pittsburgh, PA, USA

    Lauren M. DePoy, Kaitlyn A. Petersen, Kyle D. Ketchesin, Darius Becker-Krail, Madeline R. Scott & Colleen A. McClung

  3. Department of Biostatistics, University of Pittsburgh, 15261, Pittsburgh, PA, USA

    Wei Zong, RuoFei Yin & George C. Tseng

  4. Department of Human Genetics, School of Public Health, University of Pittsburgh, 15261, Pittsburgh, PA, USA

    Megan S. Perez

  5. David O. Robbins Neuroscience Program, Department of Psychology, Ohio Wesleyan University, 43015, Delaware, OH, USA

    Chelsea A. Vadnie

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Contributions

LMD, KAP, KDK, RCM, MSP, CAV, DB-K performed experiments. LMD, WZ, RY, MRS, GCT, CAM performed data analysis. LMD, WZ, RY, CAM wrote the manuscript. All authors provided manuscript edits.

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Correspondence to Colleen A. McClung.

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DePoy, L.M., Petersen, K.A., Zong, W. et al. Cell-type and sex-specific rhythmic gene expression in the nucleus accumbens. Mol Psychiatry (2024). https://doi.org/10.1038/s41380-024-02569-7

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