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Mood stabilizing drugs regulate transcription of immune, neuronal and metabolic pathway genes in Drosophila

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Abstract

Rationale

Lithium and valproate (VPA) are drugs used in the management of bipolar disorder. Even though they reportedly act on various pathways, the transcriptional targets relevant for disease mechanism and therapeutic effect remain unclear. Furthermore, multiple studies used lymphoblasts of bipolar patients as a cellular proxy, but it remains unclear whether peripheral cells provide a good readout for the effects of these drugs in the brain.

Objectives

We used Drosophila culture cells and adult flies to analyze the transcriptional effects of lithium and VPA and define mechanistic pathways.

Methods

Transcriptional profiles were determined for Drosophila S2-cells and adult fly heads following lithium or VPA treatment. Gene ontology categories were identified using the DAVID functional annotation tool with a cut-off of p < 0.05. Significantly enriched GO terms were clustered using REVIGO and DAVID functional annotation clustering. Significance of overlap between transcript lists was determined with a Fisher’s exact hypergeometric test.

Results

Treatment of cultured cells and adult flies with lithium and VPA induces transcriptional responses in genes with similar ontology, with as most prominent immune response, neuronal development, neuronal function, and metabolism.

Conclusions

(i) Transcriptional effects of lithium and VPA in Drosophila S2 cells and heads show significant overlap. (ii) The overlap between transcriptional alterations in peripheral versus neuronal cells at the single gene level is negligible, but at the gene ontology and pathway level considerable overlap can be found. (iii) Lithium and VPA act on evolutionarily conserved pathways in Drosophila and mammalian models.

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Acknowledgments

This work was funded by VIB. The authors acknowledge the collaborators of the VIB Nucleomics core for expert support with microarray experiments.

Author information

Author notes

  1. K. Hens

    Present address: Centre for Neural Circuits and Behavior, Oxford University, Oxford, UK

  2. D. Forero

    Present address: Laboratory of Neuropsychiatric Genetics, School of Medicine, Antonio Narino University, Bogota, Colombia

Authors and Affiliations

  1. Laboratory of Behavioral and Developmental Genetics, VIB-KULeuven, Herestraat 49 bus 602, 3000, Leuven, Belgium

    L. Herteleer, L. Zwarts, K. Hens, D. Forero & P. Callaerts

  2. KULeuven Department of Human Genetics, Leuven, Belgium

    L. Herteleer, L. Zwarts, K. Hens, D. Forero & P. Callaerts

  3. VIB Center for the Biology of Disease, Leuven, Belgium

    L. Herteleer, L. Zwarts, K. Hens, D. Forero & P. Callaerts

  4. Applied Molecular Genomics Group, VIB Department of Molecular Genetics, Leuven, Belgium

    D. Forero & J. Del-Favero

  5. University of Antwerp, Antwerp, Belgium

    D. Forero & J. Del-Favero

Authors
  1. L. Herteleer
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  2. L. Zwarts
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  3. K. Hens
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  6. P. Callaerts
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Corresponding author

Correspondence to P. Callaerts.

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The authors declare that they have no competing interests.

Additional information

L. Herteleer and L. Zwarts contributed equally to this work.

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Suppl. Fig. 1

Toxicity analysis of LiCl (A) Survival analysis of Berlin females using increasing doses of LiCl. (B) Survival analysis of Berlin males using increasing doses of LiCl. (C) Survival analysis of Canton-S females using increasing doses of LiCl. D) Survival analysis of Canton-S males using increasing doses of LiCl. (Logrank tests, * P < 0.05; ** P < 0.01; ***P < 0.001; **** P < 0.0001) We analyzed the effects of LiCl on two different wild type strains (Canton-S and Berlin) to rule out differential effects due to the genetic background. We examined the effects of a concentration range of 10, 15, 20, 25, 30 mM. As LiCl has been reported to be a teratogen, we administered this drug in adult flies (Dokucu et al. 2005). We observed significant decreases in survival for almost all concentrations in both Canton-S and Berlin males. Interestingly, we observed different effects on survival on females of both strains. Berlin females showed a clear increase in survival upon treatment with LiCl. Canton-S females showed to be very unresponsive to the effects of LiCl on survival. Only administration of 30 mM LiCl induced a significant decrease in survival. Based on our results, we decided to use 10 mM LiCl in the subsequent experiments. LiCl has previously been shown to have positive effects on survival in C. elegans in the concentration range that we used (McColl et al. 2008). Interestingly, these positive effects of LiCl on life span involve the downregulation of the homologue of LSD-1 (McColl et al. 2008). We show that treating S2 cells with either LiCl or VPA results in an upregulation of LSD-1 expression. Since the effects of LiCl on lifespan in C. elegans have only been investigated in a single genetic background, it will be interesting to analyze whether there are strain-specific effects as well. In addition, LiCl has been reported previously to have sex specific effects on body weight in rats (Baptista et al. 1991). A short survival study (14 days) in Drosophila did not reveal sex specific effects (Dokucu et al. 2005). In our more extensive study we do find clear sex-dependent effects. (PDF 126 kb)

Suppl. Fig. 2

Toxicity analysis of VPA (A) Survival analysis of Berlin females using increasing doses of VPA. (B) Survival analysis of Berlin males using increasing doses of VPA. (C) Survival analysis of Canton-S females using increasing doses of VPA. (D) Survival analysis of Canton-S males using increasing doses of VPA. (Logrank tests, * P < 0.05; ** P < 0.01; ***P < 0.001; **** P < 0.0001) We analyzed the effects of VPA on two different wild type strains (Canton-S and Berlin) to rule out differential effects due to the genetic background. We examined the effects of a concentration range of 0.5, 1, 3, 5, 10 mM. As VPA has been reported to be a teratogen, we administered this drug after eclosion (Dokucu et al. 2005). We observe a strong decrease in survival upon administration of increasing dosages of the drug. We can observe some variations in responsiveness between both wild type lines. Canton- S flies seem to be more sensitive to the effects of VPA on survival than Berlin flies. Based on our results, we decided to use 1 mM VPA in the subsequent experiments. (PDF 130 kb)

Suppl. Table 1

Significant transcripts in Drosophila S2-cells upon lithium treatment (FDR < 0.05, FC1.3). (PDF 594 kb)

Suppl. Table 2

Significant transcripts in Drosophila S2-cells upon VPA treatment (FDR < 0.05, FC1.3). (PDF 570 kb)

Suppl. Table 3

Enriched GO terms among significantly altered transcripts in Drosophila S2-cells and heads upon lithium and VPA treatment (P < 0.05). Terms in italics represent redundant terms summarized in one main term by making use of REVIGO (Supek et al. 2011). (PDF 84 kb)

Suppl. Table 4

DAVID: Functional Annotation Clustering of significant gene ontology terms (Enrichment >1.3). (PDF 55 kb)

Suppl. Table 5

Overlapping transcripts in Drosophila S2-cells upon lithium or VPA treatment. (PDF 365 kb)

Suppl. Table 6

Significant transcripts in Drosophila heads upon lithium treatment. (PDF 110 kb)

Suppl. Table 7

Significant transcripts in Drosophila heads upon VPA treatment. (PDF 285 kb)

Suppl. Table 8

Overlapping transcripts in Drosophila heads upon lithium or VPA treatment. (PDF 206 kb)

Suppl. Table 9

Overlapping transcripts in Drosophila S2-cells and heads upon lithium treatment. (PDF 50 kb)

Suppl. Table 10

Overlapping transcripts in Drosophila S2-cells and heads upon VPA treatment. (PDF 47 kb)

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Herteleer, L., Zwarts, L., Hens, K. et al. Mood stabilizing drugs regulate transcription of immune, neuronal and metabolic pathway genes in Drosophila . Psychopharmacology 233, 1751–1762 (2016). https://doi.org/10.1007/s00213-016-4223-z

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  • DOI: https://doi.org/10.1007/s00213-016-4223-z

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