Abstract
Brain kappa-opioid receptors (KORs) are implicated in states of motivation and emotion. Activation of KORs negatively regulates mesolimbic dopamine (DA) neurons, and KOR agonists produce depressive-like behavioral effects. To further evaluate how KOR function affects behavior, we developed mutant mice in which exon 3 of the KOR gene (Oprk1) was flanked with Cre-lox recombination (loxP) sites. By breeding these mice with lines that express Cre-recombinase (Cre) in early embryogenesis (EIIa-Cre) or only in DA neurons (dopamine transporter (DAT)-Cre), we developed constitutive KOR knockouts (KOR−/−) and conditional knockouts that lack KORs in DA-containing neurons (DAT-KORlox/lox). Autoradiography demonstrated complete ablation of KOR binding in the KOR−/− mutants, and reduced binding in the DAT-KORlox/lox mutants. Quantitative reverse transcription PCR (qPCR) studies confirmed that KOR mRNA is undetectable in the constitutive mutants and reduced in the midbrain DA systems of the conditional mutants. Behavioral characterization demonstrated that these mutant lines do not differ from controls in metrics, including hearing, vision, weight, and locomotor activity. Whereas KOR−/− mice appeared normal in the open field and light/dark box tests, DAT-KORlox/lox mice showed reduced anxiety-like behavior, an effect that is broadly consistent with previously reported effects of KOR antagonists. Sensitization to the locomotor-stimulating effects of cocaine appeared normal in KOR−/− mutants, but was exaggerated in DAT-KORlox/lox mutants. Increased sensitivity to cocaine in the DAT-KORlox/lox mutants is consistent with a role for KORs in negative regulation of DA function, whereas the lack of differences in the KOR−/− mutants suggests compensatory adaptations after constitutive receptor ablation. These mouse lines may be useful in future studies of KOR function.
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Acknowledgements
This research was supported by a National Defense Science and Engineering Graduate Fellowship (to AVV), the Frazier Medical Research Institute (to BMC), DA13429 (to L-Y L-C) and MH062366 (to WAC).
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Dr Carlezon has a US patent covering the use of kappa antagonists in the treatment of depression (Assignee: McLean Hospital). In the last 3 years Dr Carlezon has received compensation for professional services from The American College of Neuropsychopharmacology and Concert Pharmaceuticals. Dr Cohen has pending patents on pyrimidines to treat bipolar disorders, kappa-opioid agonists in bipolar mania, and on mitochondrial replacement. In the last 3 years, Dr Rudolph has provided professional services to Sunovion Pharmaceuticals and to Concert Pharmaceuticals. The other authors have no disclosures.
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Van't Veer, A., Bechtholt, A., Onvani, S. et al. Ablation of Kappa-Opioid Receptors from Brain Dopamine Neurons has Anxiolytic-Like Effects and Enhances Cocaine-Induced Plasticity. Neuropsychopharmacol 38, 1585–1597 (2013). https://doi.org/10.1038/npp.2013.58
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DOI: https://doi.org/10.1038/npp.2013.58
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