Dopamine pp 181-200

Part of the Methods in Molecular Biology book series (MIMB, volume 964) | Cite as

A Molecular Genetic Approach to Uncovering the Differential Functions of Dopamine D2 Receptor Isoforms



Alterations in the activity of the dopamine D2 receptor (D2R) have been implicated in several neurological and psychiatric disorders, including schizophrenia, Parkinson’s disease, Huntington’s disease, Tourette syndrome, attention-deficit hyperactivity disorder (ADHD), and drug addiction. Two isoforms of D2R, long form (D2LR) and short form (D2SR), have been identified. The specific function of each D2R isoform is poorly understood, primarily because isoform-selective pharmacological agents are not available. Using homologous recombination, we have generated D2LR knockout (KO) mice. D2LR KO mice are completely deficient in D2LR, but still express functional D2SR at a level similar to the total D2R level in wild-type (WT) mice. D2LR is generally the predominant isoform expressed in WT mice. We showed that D2LR KO mice displayed a number of robust behavioral phenotypes distinct from WT mice, indicating that D2LR and D2SR have differential functions. In this chapter we describe the generation and characterization of the D2LR KO mouse. This genetic approach provides a valuable research tool to investigate the functional role of individual D2R isoforms in the mammalian central nervous system (CNS).

Key words

Dopamine D2 receptor Knockout mouse Homologous recombination Transfection of embryonic stem cell Genotyping 


  1. 1.
    Dal Toso R, Sommer B, Ewert M, Herb A, Pritchett DB, Bach A, Shivers BD, Seeburg PH (1989) The dopamine D2 receptor: two molecular forms generated by alternative splicing. EMBO J 8:4025–4034PubMedGoogle Scholar
  2. 2.
    Giros B, Sokoloff P, Martres MP, Riou JF, Emorine LJ, Schwartz JC (1989) Alternative splicing directs the expression of two D2 dopamine receptor isoforms. Nature 342:923–926PubMedCrossRefGoogle Scholar
  3. 3.
    Monsma FJ Jr, McVittie LD, Gerfen CR, Mahan LC, Sibley DR (1989) Multiple D2 dopamine receptors produced by alternative RNA splicing. Nature 342:926–929PubMedCrossRefGoogle Scholar
  4. 4.
    Chio CL, Hess GF, Graham RS, Huff RM (1990) A second molecular form of D2 dopamine receptor in rat and bovine caudate nucleus. Nature 343:266–269PubMedCrossRefGoogle Scholar
  5. 5.
    Wang Y, Xu R, Sasaoka T, Tonegawa S, Kung M-P, Sankoorikal E-B (2000) Dopamine D2 long receptor-deficient mice display alterations in striatum-dependent functions. J Neurosci 20:8305–8314PubMedGoogle Scholar
  6. 6.
    Usiello A, Baik J-H, Rouge-Pont F, Picetti R, Dierich A, LeMeur M, Piazza PV, Borrelli E (2000) Distinct functions of the two isoforms of dopamine D2 receptors. Nature 408:199–203PubMedCrossRefGoogle Scholar
  7. 7.
    Smith JW, Fetsko LA, Xu R, Wang Y (2002) Dopamine D2L receptor knockout mice display deficits in positive and negative reinforcing properties of morphine and in avoidance learning. Neuroscience 113:755–765PubMedCrossRefGoogle Scholar
  8. 8.
    Xu R, Hranilovic D, Fetsko LA, Bucan M, Wang Y (2002) Dopamine D2S and D2L receptors may differentially contribute to the actions of antipsychotic and psychotic agents in mice. Mol Psychiatry 7:1075–1082PubMedCrossRefGoogle Scholar
  9. 9.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  10. 10.
    Mombaerts P, Clarke AR, Hooper ML, Tonegawa S (1991) Creation of a large genomic deletion at the T-cell antigen receptor beta-subunit locus in mouse embryonic stem cells by gene targeting. Proc Natl Acad Sci USA 88:3084–3087PubMedCrossRefGoogle Scholar
  11. 11.
    Adra CN, Boer PH, McBurney MW (1987) Cloning and expression of the mouse pgk-1 gene and the nucleotide sequence of its promoter. Gene 60:65–74PubMedCrossRefGoogle Scholar
  12. 12.
    Ausubel F, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1995) Short protocols in molecular biology, 3rd edn. Wiley, New YorkGoogle Scholar
  13. 13.
    Nagy A, Gertsenstein M, Vintersten K, Behringer R (2003) Manipulating the mouse embryo: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  14. 14.
    Matise M, Auerbach W, Joyner AL (2000) Production of targeted embryonic stem cell clones. In: Joyner A (ed) Gene targeting: a practical approach, 2nd edn. Oxford University Press, OxfordGoogle Scholar
  15. 15.
    Papaioannou V, Johnson R (2000) Production of chimeras by blastocyst and morula injection of targeted ES cells. In: Joyner A (ed) Gene targeting: a practice approach, 2nd edn. Oxford University Press, OxfordGoogle Scholar
  16. 16.
    Dang MT, Yokoi F, Yin HH, Lovinger DM, Wang Y, Li Y (2006) Disrupted motor learning and long-term synaptic plasticity in mice lacking NMDAR1 in the striatum. Proc Natl Acad Sci USA 103:15254–15259PubMedCrossRefGoogle Scholar
  17. 17.
    te Riele H, Maandag ER, Berns A (1992) Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc Natl Acad Sci USA 89:5128–5132CrossRefGoogle Scholar
  18. 18.
    Thomas KR, Deng C, Capecchi MR (1992) High-fidelity gene targeting in embryonic stem cells by using sequence replacement vectors. Mol Cell Biol 12:2919–2923PubMedGoogle Scholar
  19. 19.
    Smith GR (1994) Hotspots of homologous recombination. Experientia 50:234–241PubMedCrossRefGoogle Scholar
  20. 20.
    Sasaoka T, Imamura M, Araishi K, Noguchi S, Mizuno Y, Takagoshi N, Hama H, Wakabayashi-Takai E, Yoshimoto-Matsuda Y, Nonaka I, Kaneko K, Yoshida M, Ozawa E (2003) Pathological analysis of muscle hypertrophy and degeneration in muscular dystrophy in gamma-sarcoglycan-deficient mice. Neuromuscul Disord 13:193–206PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Department of Pharmacology, College of Medicine, Beckman Institute for Advanced Science and TechnologyUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Department of Laboratory Animal ScienceKitasato University School of MedicineKanagawaJapan
  3. 3.Department of NeurologyHospital of University of PennsylvaniaPhiladelphiaUSA

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