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siRNA-mediated knockdown of the serotonin transporter in the adult mouse brain

Molecular Psychiatry volume 10, pages 782–789 (2005)Cite this article

Abstract

Selective serotonin reuptake inhibitors (SSRIs) are widely used antidepressant drugs that increase the extracellular levels of serotonin by blocking the reuptake activity of the serotonin transporter (SERT). Although SSRIs elevate brain serotonergic neurotransmission acutely, their full therapeutic effects involve neurochemical adaptations that emerge following chronic drug administration. The adaptive downregulation of SERT has recently been implicated in the therapeutic response of SSRIs. Interestingly, studies using SERT-knockout mice reveal somewhat paradoxical depression-related effects, probably specific to the downregulation of SERT during early development. However, the behavioral significance of SSRI-mediated downregulation of SERT during adulthood is still unknown. We investigated whether somatic gene manipulation, triggered by infusing short interfering RNA (siRNA) into the ventricular system, would enable the downregulation of SERT in the adult mouse brain. Infusing the SERT-targeting siRNA, for 2 weeks, significantly reduced the mRNA levels of SERT in raphe nuclei. Further, a significant, specific and widespread downregulation of SERT-binding sites was achieved in the brain. In contrast, 2-week infusion of the SSRI, citalopram, produced a widespread downregulation of SERT-binding sites, independent of any alterations at the mRNA level. Irrespective of their mechanisms for downregulating SERT in the brain, infusions of SERT-siRNA or citalopram elicited a similar antidepressant-related behavioral response in the forced swim test. These results signify a role for the downregulation of SERT in mediating the antidepressant action of SSRIs in adults. Further, these data demonstrate that siRNA-induced widespread knockdown of gene expression serves as a powerful tool for assessing the function of endogenous genes in the adult brain.

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Acknowledgements

We gratefully acknowledge Sabine Leonhard, Hugo Bürki, Dominique Fehlmann, Dora Khar, David Kirk and Jose Crespo for their excellent technical assistance, and Dr Graeme Bilbe (Head of Neuroscience Research, Novartis Institutes of BioMedical Sciences) for his continuous support of our RNAi projects.

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Authors and Affiliations

  1. Psychiatry Program, Neuroscience Research, Novartis Institutes for BioMedical Research, Novartis Pharma AG, Basel, Switzerland

    D R Thakker, H van der Putten, R Maier, D Hoyer & J F Cryan

  2. Functional Genomics, Novartis Institutes for BioMedical Research, Novartis Pharma AG, Basel, Switzerland

    F Natt & D Hüsken

  3. Department of Neuropharmacology, The Harold L Dorris Neurological Research Center, The Scripps Research Institute, La Jolla, CA, USA

    D Hoyer

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  1. D R Thakker
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  2. F Natt
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Correspondence to J F Cryan.

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Thakker, D., Natt, F., Hüsken, D. et al. siRNA-mediated knockdown of the serotonin transporter in the adult mouse brain. Mol Psychiatry 10, 782–789 (2005). https://doi.org/10.1038/sj.mp.4001687

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