Abstract
Dys-regulation of the dopamine system is strongly implicated in schizophrenia and addiction, and the dopamine D2 receptor (DRD2) is a critical regulator of dopaminergic activity and mediates dopamine actions on motivation and reward. We have identified a single nucleotide polymorphism in the DRD2 gene (rs2734836) that reduces the binding of the transcriptional repressor Freud-1/CC2D1A and in turn alters expression of the dopamine D2 receptor (Rogaeva et al., J Biol Chem 282:20897–20905, 2007). Increasing evidence shows that promoter methylation and demethylation during the early postnatal critical period can establish lifelong change in hippocampal glucocorticoid receptor expression and stress reactivity (Meaney and Szyf, Trends Neurosci 28:456–463, 2005). The early postnatal period is also that time during which expression of the DRD2 matures and reaches adult levels. Hence, we hypothesize that differences in DNA methylation of the DRD2 promoter may correlate with altered expression of the receptor in adults, and predispose to schizophrenia. In this chapter, we review methods for both genotype analysis of key polymorphisms of the DRD2 gene (rs2734836 and the Taq1A variant rs1800497), as well as methodology for analysis of the DNA methylation. These methods can be easily adapted to any gene of interest using the methods for oligonucleotide primer selection and PCR conditions suggested.
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Acknowledgments
PRA was supported by funding from the Canadian Institutes of Health Research, Ontario Mental Health Foundation, and Heart and Stroke Foundation Canadian Partnership for Stroke Recovery. LMF was supported by postdoctoral funding from the National Science and Engineering Research Council of Canada.
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Fiori, L.M., Daigle, M., Albert, P.R. (2015). Genetic and Epigenetic Methods for Analysis of the Dopamine D2 Receptor Gene. In: Tiberi, M. (eds) Dopamine Receptor Technologies. Neuromethods, vol 96. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2196-6_1
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DOI: https://doi.org/10.1007/978-1-4939-2196-6_1
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