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Dosage sensitivity intolerance of VIPR2 microduplication is disease causative to manifest schizophrenia-like phenotypes in a novel BAC transgenic mouse model

Abstract

Recent genome-wide association studies (GWAS) have identified copy number variations (CNVs) at chromosomal locus 7q36.3 that significantly contribute to the risk of schizophrenia, with all of the microduplications occurring within a single gene: vasoactive intestinal peptide receptor 2 (VIPR2). To confirm disease causality and translate such a genetic vulnerability into mechanistic and pathophysiological insights, we have developed a series of conditional VIPR2 bacterial artificial chromosome (BAC) transgenic mouse models of VIPR2 CNV. VIPR2 CNV mouse model recapitulates gene expression and signaling deficits seen in human CNV carriers. VIPR2 microduplication in mice elicits prominent dorsal striatal dopamine dysfunction, cognitive, sensorimotor gating, and social behavioral deficits preceded by an increase of striatal cAMP/PKA signaling and the disrupted early postnatal striatal development. Genetic removal of VIPR2 transgene expression via crossing with Drd1a-Cre BAC transgenic mice rescued the dopamine D2 receptor abnormality and multiple behavioral deficits, implicating a pathogenic role of VIPR2 overexpression in dopaminoceptive neurons. Thus, our results provide further evidence to support the GWAS studies that the dosage sensitivity intolerance of VIPR2 is disease causative to manifest schizophrenia-like dopamine, cognitive, and social behavioral deficits in mice. The conditional BAC transgenesis offers a novel strategy to model CNVs with a gain-of -copies and facilitate the genetic dissection of when/where/how the genetic vulnerabilities affect development, structure, and function of neural circuits. Our findings have important implications for therapeutic development, and the etiology-relevant mouse model provides a useful preclinical platform for drug discovery.

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References

  1. Feuk L, Carson AR, Scherer SW. Structural variation in the human genome. Nat Rev Genet. 2006;7:85–97.

    Article  CAS  PubMed  Google Scholar 

  2. Malhotra D, Sebat J. CNVs: harbingers of a rare variant revolution in psychiatric genetics. Cell. 2012;148:1223–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Rice AM, McLysaght A. Dosage sensitivity is a major determinant of human copy number variant pathogenicity. Nat Commun. 2017;8:14366.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Andrews T, Honti F, Pfundt R, de Leeuw N, Hehir-Kwa J, Vulto-van Silfhout A, et al. The clustering of functionally related genes contributes to CNV-mediated disease. Genome Res. 2015;25:802–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Rippey C, Walsh T, Gulsuner S, Brodsky M, Nord AS, Gasperini M, et al. Formation of chimeric genes by copy-number variation as a mutational mechanism in schizophrenia. Am J Hum Genet. 2013;93:697–710.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Nestler EJ, Hyman SE. Animal models of neuropsychiatric disorders. Nat Neurosci. 2010;13:1161–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Vacic V, McCarthy S, Malhotra D, Murray F, Chou HH, Peoples A, et al. Duplications of the neuropeptide receptor gene VIPR2 confer significant risk for schizophrenia. Nature. 2011;471:499–503.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Levinson DF, Duan J, Oh S, Wang K, Sanders AR, Shi J, et al. Copy number variants in schizophrenia: confirmation of five previous findings and new evidence for 3q29 microdeletions and VIPR2 duplications. Am J Psychiatry. 2011;168:302–16.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Li Z, Chen J, Xu Y, Yi Q, Ji W, Wang P, et al. Genome-wide analysis of the role of copy number variation in schizophrenia risk in Chinese. Biol Psychiatry. 2016;80:331–7.

    Article  PubMed  Google Scholar 

  10. Cnv, Schizophrenia Working Groups of the Psychiatric Genomics C, Psychosis Endophenotypes International C. Contribution of copy number variants to schizophrenia from a genome-wide study of 41,321 subjects. Nat Genet. 2017;49:27–35.

    Article  CAS  Google Scholar 

  11. Harmar AJ, Fahrenkrug J, Gozes I, Laburthe M, May V, Pisegna JR, et al. Pharmacology and functions of receptors for vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide: IUPHAR review 1. Br J Pharmacol. 2012;166:4–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Hill JM. Vasoactive intestinal peptide in neurodevelopmental disorders: therapeutic potential. Curr Pharm Des. 2007;13:1079–89.

    Article  CAS  PubMed  Google Scholar 

  13. Ago Y, Condro MC, Tan YV, Ghiani CA, Colwell CS, Cushman JD, et al. Reductions in synaptic proteins and selective alteration of prepulse inhibition in male C57BL/6 mice after postnatal administration of a VIP receptor (VIPR2) agonist. Psychopharmacology. 2015;232:2181–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Quik M, Iversen LL, Bloom SR. Effect of vasoactive intestinal peptide (VIP) and other peptides on cAMP accumulation in rat brain. Biochem Pharmacol. 1978;27:2209–13.

    Article  CAS  PubMed  Google Scholar 

  15. Weiss S, Sebben M, Kemp DE, Bockaert J. Vasoactive intestinal peptide actions on cyclic AMP levels in cultured striatal neurons. Peptides. 1986;7 Suppl 1:187–92.

    Article  CAS  PubMed  Google Scholar 

  16. Girault JA, Shalaby IA, Rosen NL, Greengard P. Regulation by cAMP and vasoactive intestinal peptide of phosphorylation of specific proteins in striatal cells in culture. Proc Natl Acad Sci USA. 1988;85:7790–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Girault JA, Horiuchi A, Gustafson EL, Rosen NL, Greengard P. Differential expression of ARPP-16 and ARPP-19, two highly related cAMP-regulated phosphoproteins, one of which is specifically associated with dopamine-innervated brain regions. J Neurosci. 1990;10:1124–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Greengard P, Allen PB, Nairn AC. Beyond the dopamine receptor: the DARPP-32/protein phosphatase-1 cascade. Neuron. 1999;23:435–47.

    Article  CAS  PubMed  Google Scholar 

  19. Taylor DP, Pert CB. Vasoactive intestinal polypeptide: specific binding to rat brain membranes. Proc Natl Acad Sci USA. 1979;76:660–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Vertongen P, Schiffmann SN, Gourlet P, Robberecht P. Autoradiographic visualization of the receptor subclasses for vasoactive intestinal polypeptide (VIP) in rat brain. Peptides. 1997;18:1547–54.

    Article  CAS  PubMed  Google Scholar 

  21. McCutcheon RA, Abi-Dargham A, Howes OD. Schizophrenia, dopamine and the striatum: from biology to symptoms. Trends Neurosci. 2019; 42(3): 205–220.

  22. Kesby JP, Eyles DW, McGrath JJ, Scott JG. Dopamine, psychosis and schizophrenia: the widening gap between basic and clinical neuroscience. Transl Psychiatry. 2018;8:30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Simpson EH, Kellendonk C, Kandel E. A possible role for the striatum in the pathogenesis of the cognitive symptoms of schizophrenia. Neuron. 2010;65:585–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Kellendonk C, Simpson EH, Polan HJ, Malleret G, Vronskaya S, Winiger V, et al. Transient and selective overexpression of dopamine D2 receptors in the striatum causes persistent abnormalities in prefrontal cortex functioning. Neuron. 2006;49:603–15.

    Article  CAS  PubMed  Google Scholar 

  25. Goto A, Nakahara I, Yamaguchi T, Kamioka Y, Sumiyama K, Matsuda M, et al. Circuit-dependent striatal PKA and ERK signaling underlies rapid behavioral shift in mating reaction of male mice. Proc Natl Acad Sci USA. 2015;112:6718–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kozorovitskiy Y, Peixoto R, Wang W, Saunders A, Sabatini BL. Neuromodulation of excitatory synaptogenesis in striatal development. Elife 2015;4:10111.

  27. Shneider Y, Shtrauss Y, Yadid G, Pinhasov A. Differential expression of PACAP receptors in postnatal rat brain. Neuropeptides. 2010;44:509–14.

    Article  CAS  PubMed  Google Scholar 

  28. Usdin TB, Bonner TI, Mezey E. Two receptors for vasoactive intestinal polypeptide with similar specificity and complementary distributions. Endocrinology. 1994;135:2662–80.

    Article  CAS  PubMed  Google Scholar 

  29. Sheward WJ, Lutz EM, Harmar AJ. The distribution of vasoactive intestinal peptide2 receptor messenger RNA in the rat brain and pituitary gland as assessed by in situ hybridization. Neuroscience. 1995;67:409–18.

    Article  CAS  PubMed  Google Scholar 

  30. Michel MC, Wieland T, Tsujimoto G. How reliable are G-protein-coupled receptor antibodies? Naunyn-Schmiede’s Arch Pharmacol. 2009;379:385–8.

    Article  CAS  Google Scholar 

  31. An S, Tsai C, Ronecker J, Bayly A, Herzog ED. Spatiotemporal distribution of vasoactive intestinal polypeptide receptor 2 in mouse suprachiasmatic nucleus. J Comp Neurol. 2012;520:2730–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Miller JE, Granados-Fuentes D, Wang T, Marpegan L, Holy TE, Herzog ED. Vasoactive intestinal polypeptide mediates circadian rhythms in mammalian olfactory bulb and olfaction. J Neurosci. 2014;34:6040–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gerfen CR, Paletzki R, Heintz N. GENSAT BAC cre-recombinase driver lines to study the functional organization of cerebral cortical and basal ganglia circuits. Neuron. 2013;80:1368–83.

    Article  CAS  PubMed  Google Scholar 

  34. Harris JA, Hirokawa KE, Sorensen SA, Gu H, Mills M, Ng LL, et al. Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front Neural Circuits. 2014;8:76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Schulz S, Mann A, Novakhov B, Piggins HD, Lupp A. VPAC2 receptor expression in human normal and neoplastic tissues: evaluation of the novel MAB SP235. Endocr Connect. 2015;4:18–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Schulz S, Rocken C, Mawrin C, Weise W, Hollt V, Schulz S. Immunocytochemical identification of VPAC1, VPAC2, and PAC1 receptors in normal and neoplastic human tissues with subtype-specific antibodies. Clin Cancer Res. 2004;10:8235–42.

    Article  CAS  PubMed  Google Scholar 

  37. Tamura M, Mukai J, Gordon JA, Gogos JA. Developmental inhibition of Gsk3 rescues behavioral and neurophysiological deficits in a mouse model of schizophrenia predisposition. Neuron. 2016;89:1100–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Mena A, Ruiz-Salas JC, Puentes A, Dorado I, Ruiz-Veguilla M, De la Casa LG, et al. Reduced prepulse inhibition as a biomarker of schizophrenia. Front Behav Neurosci. 2016;10:202.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Valsamis B, Schmid S. Habituation and prepulse inhibition of acoustic startle in rodents. J Vis Exp. 2011;55:e3446.

    Google Scholar 

  40. Kaidanovich-Beilin O, Lipina T, Vukobradovic I, Roder J, Woodgett JR. Assessment of social interaction behaviors. J Vis Exp. 2011: 2473.

  41. Hiroi N, Zhu H, Lee M, Funke B, Arai M, Itokawa M, et al. A 200-kb region of human chromosome 22q11.2 confers antipsychotic-responsive behavioral abnormalities in mice. Proc Natl Acad Sci USA. 2005;102:19132–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Yang XW, Gong S. An overview on the generation of BAC transgenic mice for neuroscience research. Curr. Protoc. Neurosci. 2005; Chapter 5: 5.20.1–5.20.11.

  43. Stojanovic T, Orlova M, Sialana FJ, Hoger H, Stuchlik S, Milenkovic I, et al. Validation of dopamine receptor DRD1 and DRD2 antibodies using receptor deficient mice. Amino Acids. 2017;49:1101–9.

    Article  CAS  PubMed  Google Scholar 

  44. Yin HH, Knowlton BJ. Contributions of striatal subregions to place and response learning. Learn Mem. 2004;11:459–63.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Laruelle M. Imaging dopamine transmission in schizophrenia. A review and meta-analysis. Q J Nucl Med. 1998;42:211–21.

    CAS  PubMed  Google Scholar 

  46. Kulkosky PJ, Doyle JS, Cook VI, Glazner GW, Foderaro MA. Vasoactive intestinal peptide: behavioral effects in the rat and hamster. Pharmacol, Biochem, Behav. 1989;34:387–93.

    Article  CAS  Google Scholar 

  47. Akhmedov D, Mendoza-Rodriguez MG, Rajendran K, Rossi M, Wess J, Berdeaux R. Gs-DREADD knock-in mice for tissue-specific, temporal stimulation of cyclic AMP signaling. Mol Cell Biol. 2017;37:e00584–16.

  48. Wojcik SM, Rhee JS, Herzog E, Sigler A, Jahn R, Takamori S, et al. An essential role for vesicular glutamate transporter 1 (VGLUT1) in postnatal development and control of quantal size. Proc Natl Acad Sci USA. 2004;101:7158–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Nakamura KC, Fujiyama F, Furuta T, Hioki H, Kaneko T. Afferent islands are larger than mu-opioid receptor patch in striatum of rat pups. Neuroreport. 2009;20:584–8.

    Article  CAS  PubMed  Google Scholar 

  50. Lu XH. Genetically-directed sparse neuronal labeling in BAC transgenic mice through mononucleotide repeat frameshift. Sci Rep. 2017;7:43915.

  51. Caille I, Dumartin B, Le Moine C, Begueret J, Bloch B. Ontogeny of the D1 dopamine receptor in the rat striatonigral system: an immunohistochemical study. Eur J Neurosci. 1995;7:714–22.

    Article  CAS  PubMed  Google Scholar 

  52. Goldman-Rakic PS, Castner SA, Svensson TH, Siever LJ, Williams GV. Targeting the dopamine D1 receptor in schizophrenia: insights for cognitive dysfunction. Psychopharmacology. 2004;174:3–16.

    Article  CAS  PubMed  Google Scholar 

  53. Rodrigues S, Salum C, Ferreira TL. Dorsal striatum D1-expressing neurons are involved with sensorimotor gating on prepulse inhibition test. J Psychopharmacol. 2017; https://doi.org/10.1177/0269881116686879.

  54. Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci. 2010;13:133–40.

    Article  CAS  PubMed  Google Scholar 

  55. Nelson KB, Grether JK, Croen LA, Dambrosia JM, Dickens BF, Jelliffe LL, et al. Neuropeptides and neurotrophins in neonatal blood of children with autism or mental retardation. Ann Neurol. 2001;49:597–606.

    Article  CAS  PubMed  Google Scholar 

  56. Liu X, Li Z, Fan C, Zhang D, Chen J. Genetics implicate common mechanisms in autism and schizophrenia: synaptic activity and immunity. J Med Genet. 2017;54:511–20.

    CAS  PubMed  Google Scholar 

  57. Stefansson H, Meyer-Lindenberg A, Steinberg S, Magnusdottir B, Morgen K, Arnarsdottir S, et al. CNVs conferring risk of autism or schizophrenia affect cognition in controls. Nature. 2014;505:361–6.

    Article  CAS  PubMed  Google Scholar 

  58. Birnbaum R, Weinberger DR. Genetic insights into the neurodevelopmental origins of schizophrenia. Nat Rev Neurosci. 2017;18:727–40.

    Article  CAS  PubMed  Google Scholar 

  59. Wang N, Gray M, Lu XH, Cantle JP, Holley SM, Greiner E, et al. Neuronal targets for reducing mutant huntingtin expression to ameliorate disease in a mouse model of Huntington’s disease. Nat Med. 2014;20:536–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Arnsten AF. Catecholamine influences on dorsolateral prefrontal cortical networks. Biol Psychiatry. 2011;69:e89–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Kelly MP, Stein JM, Vecsey CG, Favilla C, Yang X, Bizily SF, et al. Developmental etiology for neuroanatomical and cognitive deficits in mice overexpressing Galphas, a G-protein subunit genetically linked to schizophrenia. Mol Psychiatry. 2009;14:398–415, 347.

    Article  CAS  PubMed  Google Scholar 

  62. Moyer CE, Shelton MA, Sweet RA. Dendritic spine alterations in schizophrenia. Neuroscience Lett. 2014;601:46–53.

  63. Kozorovitskiy Y, Saunders A, Johnson CA, Lowell BB, Sabatini BL. Recurrent network activity drives striatal synaptogenesis. Nature. 2012;485:646–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Kegeles LS, Abi-Dargham A, Frankle WG, Gil R, Cooper TB, Slifstein M, et al. Increased synaptic dopamine function in associative regions of the striatum in schizophrenia. Arch Gen Psychiatry. 2010;67:231–9.

    Article  CAS  PubMed  Google Scholar 

  65. Howes OD, Williams M, Ibrahim K, Leung G, Egerton A, McGuire PK, et al. Midbrain dopamine function in schizophrenia and depression: a post-mortem and positron emission tomographic imaging study. Brain. 2013;136:3242–51.

    Article  PubMed  PubMed Central  Google Scholar 

  66. Brunelin J, Fecteau S, Suaud-Chagny MF. Abnormal striatal dopamine transmission in schizophrenia. Curr Med Chem. 2013;20:397–404.

    CAS  PubMed  PubMed Central  Google Scholar 

  67. Balleine BW, Delgado MR, Hikosaka O. The role of the dorsal striatum in reward and decision-making. J Neurosci. 2007;27:8161–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Yin HH, Knowlton BJ. The role of the basal ganglia in habit formation. Nat Rev Neurosci. 2006;7:464–76.

    Article  CAS  PubMed  Google Scholar 

  69. Howes OD. Elevated striatal dopamine function linked to prodromal signs of schizophrenia. Arch Gen Psychiatry. 2009;66:13–20.

    Article  PubMed  Google Scholar 

  70. Balleine BW, O’Doherty JP. Human and rodent homologies in action control: corticostriatal determinants of goal-directed and habitual action. Neuropsychopharmacol. 2010;35:48–69.

    Article  Google Scholar 

  71. Baez-Mendoza R, Schultz W. The role of the striatum in social behavior. Front Neurosci. 2013;7:233.

    Article  PubMed  PubMed Central  Google Scholar 

  72. Gold JM, Strauss GP, Waltz JA, Robinson BM, Brown JK, Frank MJ. Negative symptoms of schizophrenia are associated with abnormal effort-cost computations. Biol Psychiatry. 2013;74:130–6.

    Article  PubMed  PubMed Central  Google Scholar 

  73. Morris RW, Quail S, Griffiths KR, Green MJ, Balleine BW. Corticostriatal control of goal-directed action is impaired in schizophrenia. Biol Psychiatry. 2015;77:187–95.

    Article  PubMed  Google Scholar 

  74. Chun S, Westmoreland JJ, Bayazitov IT, Eddins D, Pani AK, Smeyne RJ, et al. Specific disruption of thalamic inputs to the auditory cortex in schizophrenia models. Science. 2014;344:1178–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Chu A, Caldwell JS, Chen YA. Identification and characterization of a small molecule antagonist of human VPAC(2) receptor. Mol Pharm. 2010;77:95–101.

    Article  CAS  Google Scholar 

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Acknowledgements

The project was supported by a NARSAD young investigator award from the Brain & Behavior Research Foundation and a Center for Excellence in Arthritis and Rheumatology grant from LSUHSC to X-HL. X-HL conceived the idea, designed the experiments, and wrote the manuscript. XWY provided valuable guidance in the study, assisting in interpreting the findings, and modifying the manuscript. XWY and NEG served as mentors for the NARSAD award. XT performed most of the behavioral and pathologic studies in Figs. 1–5 and Supplementary Figs. S2–12. BL at LSUHSC performed the HPLC experiment. AR, MWE-S, AB, IVS, and KH performed some behavioral and pathologic studies. Dr Rebecca Berdeaux from the Department of Integrative Biology and Pharmacology, McGovern Medical School at The University of Texas Health Science Center at Houston generously provided CRE-LUC knockin mice. RLK provided open filed, rotarod behavioral equipment and synaptophysin antibody.

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Tian, X., Richard, A., El-Saadi, M.W. et al. Dosage sensitivity intolerance of VIPR2 microduplication is disease causative to manifest schizophrenia-like phenotypes in a novel BAC transgenic mouse model. Mol Psychiatry 24, 1884–1901 (2019). https://doi.org/10.1038/s41380-019-0492-3

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