, Volume 236, Issue 9, pp 2687–2697 | Cite as

Modulation by chronic antipsychotic administration of PKA- and GSK3β-mediated pathways and the NMDA receptor in rat ventral midbrain

  • Bo Pan
  • Chao DengEmail author
Original Investigation



Antipsychotics exert therapeutic effects by modulating various cellular signalling pathways and several types of receptors, including PKA- and GSK3β-mediated signalling pathways, and NMDA receptors. The ventral midbrain, mainly containing the ventral tegmental area (VTA) and substantia nigra (SN), are the nuclei with dopamine origins in the brain, which are also involved in the actions of antipsychotics. Whether antipsychotics can modulate these cellular pathways in the ventral midbrain is unknown.


This study aims to investigate the effects of antipsychotics, including aripiprazole (a dopamine D2 receptor (D2R) partial agonist), bifeprunox (a D2R partial agonist), and haloperidol (a D2R antagonist) on the PKA- and GSK3β-mediated pathways and NMDA receptors in the ventral midbrain.


Male rats were orally administered aripiprazole (0.75 mg/kg, t.i.d. (ter in die)), bifeprunox (0.8 mg/kg, t.i.d.), haloperidol (0.1 mg/kg, t.i.d.) or vehicle for either 1 week or 10 weeks. The levels of PKA, p-PKA, Akt, p-Akt, GSK3β, p-GSK3β, Dvl-3, β-catenin, and NMDA receptor subunits in the ventral midbrain were assessed by Western Blots.


The results showed that chronic antipsychotic treatment with aripiprazole selectively increased PKA activity in the VTA. Additionally, all three drugs elevated the activity of the Akt–GSK3β signalling pathway in a time-dependent manner, while only aripiprazole stimulated the Dvl-3–GSK3β–β-catenin signalling pathway in the SN. Furthermore, chronic administration with both aripiprazole and haloperidol decreased the expression of NMDA receptors.


This study suggests that activating PKA- and GSK3β-mediated pathways and downregulating NMDA receptor expression in the ventral midbrain might contribute to the clinical effects of antipsychotics.


Antipsychotics Ventral midbrain PKA GSK3β NMDA receptor 



We would like to thank Dr. Jiamei Lian and Dr. Michael De-Santis for their technical assistance.

Funding information

This study was supported by the Australian National Health and Medical Research Council project grant (APP1008473) to Chao Deng. Bo Pan was supported by the Natural Science Foundation of the Higher Education Institutions of Jiangsu Province, China (17KJB310018), the China Postdoctoral Science Foundation (2018 M632401), and the Natural Science Foundation of Jiangsu Province of China (BK20171290).

Compliance with ethical standards

All experimental procedures were approved by the Animal Ethics Committee (AE11/02) of the University of Wollongong and complied with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes (2004).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. (Administration) FUSFaD (2005) Guidance for industry on estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers. In: Services USDoHaH, Administration FaD, (CDER) CfDEaR (eds) Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, Rockville, Maryland, USAGoogle Scholar
  2. Alimohamad H, Rajakumar N, Seah YH, Rushlow W (2005a) Antipsychotics alter the protein expression levels of beta-catenin and GSK-3 in the rat medial prefrontal cortex and striatum. Biol Psychiatry 57:533–542CrossRefGoogle Scholar
  3. Alimohamad H, Sutton L, Mouyal J, Rajakumar N, Rushlow WJ (2005b) The effects of antipsychotics on beta-catenin, glycogen synthase kinase-3 and dishevelled in the ventral midbrain of rats. J Neurochem 95:513–525CrossRefGoogle Scholar
  4. Allen JA, Yost JM, Setola V, Chen X, Sassano MF, Chen M, Peterson S, Yadav PN, Huang XP, Feng B, Jensen NH, Che X, Bai X, Frye SV, Wetsel WC, Caron MG, Javitch JA, Roth BL, Jin J (2011) Discovery of beta-arrestin-biased dopamine D2 ligands for probing signal transduction pathways essential for antipsychotic efficacy. Proc Natl Acad Sci U S A 108:18488–18493CrossRefGoogle Scholar
  5. Assie MB, Dominguez H, Consul-Denjean N, Newman-Tancredi A (2006) In vivo occupancy of dopamine D2 receptors by antipsychotic drugs and novel compounds in the mouse striatum and olfactory tubercles. Naunyn Schmiedeberg’s Arch Pharmacol 373:441–450CrossRefGoogle Scholar
  6. Beaulieu JM, Gainetdinov RR (2011) The physiology, signaling, and pharmacology of dopamine receptors. Pharmacol Rev 63:182–217CrossRefGoogle Scholar
  7. Beaulieu JM, Gainetdinov RR, Caron MG (2009) Akt/GSK3 signaling in the action of psychotropic drugs. Annu Rev Pharmacol Toxicol 49:327–347CrossRefGoogle Scholar
  8. Beier KT, Steinberg EE, DeLoach KE, Xie S, Miyamichi K, Schwarz L, Gao XJ, Kremer EJ, Malenka RC, Luo L (2015) Circuit architecture of VTA dopamine neurons revealed by systematic input-output mapping. Cell 162:622–634CrossRefGoogle Scholar
  9. Boyd PJ, Cunliffe VT, Roy S, Wood JD (2015) Sonic hedgehog functions upstream of disrupted-in-schizophrenia 1 (disc1): implications for mental illness. Biol Open 4:1336–1343CrossRefGoogle Scholar
  10. Carlsson A, Lindqvist M (1963) Effect of chlorpromazine or haloperidol on formation of 3-methoxytyramine and normetanephrine in mouse brain. Acta Pharmacol Toxicol (Copenh) 20:140–144CrossRefGoogle Scholar
  11. Casey DE, Sands EE, Heisterberg J, Yang HM (2008) Efficacy and safety of bifeprunox in patients with an acute exacerbation of schizophrenia: results from a randomized, double-blind, placebo-controlled, multicenter, dose-finding study. Psychopharmacology 200:317–331CrossRefGoogle Scholar
  12. Correll CU (2010) From receptor pharmacology to improved outcomes: individualising the selection, dosing, and switching of antipsychotics. Eur Psychiatry 25(Supplement 2):S12–S21CrossRefGoogle Scholar
  13. 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 8:4025–4034CrossRefGoogle Scholar
  14. Dwivedi Y, Rizavi HS, Pandey GN (2002) Differential effects of haloperidol and clozapine on [(3)H]cAMP binding, protein kinase A (PKA) activity, and mRNA and protein expression of selective regulatory and catalytic subunit isoforms of PKA in rat brain. J Pharmacol Exp Ther 301:197–209CrossRefGoogle Scholar
  15. El Hage C, Bédard A-M, Samaha A-N (2015) Antipsychotic treatment leading to dopamine supersensitivity persistently alters nucleus accumbens function. Neuropharmacology 99:715–725CrossRefGoogle Scholar
  16. el Mestikawy S, Hamon M (1986) Is dopamine-induced inhibition of adenylate cyclase involved in the autoreceptor-mediated negative control of tyrosine hydroxylase in striatal dopaminergic terminals? J Neurochem 47:1425–1433CrossRefGoogle Scholar
  17. Emamian ES (2012) AKT/GSK3 signaling pathway and schizophrenia. Front Mol Neurosci 5:33CrossRefGoogle Scholar
  18. Ford CP (2014) The role of D2-autoreceptors in regulating dopamine neuron activity and transmission. Neuroscience 282C:13–22CrossRefGoogle Scholar
  19. Gardner DM, Murphy AL, O'Donnell H, Centorrino F, Baldessarini RJ (2010) International consensus study of antipsychotic dosing. Am J Psychiatry 167:686–693CrossRefGoogle Scholar
  20. Ginovart N, Kapur S (2012) Role of dopamine D(2) receptors for antipsychotic activity. Handb Exp Pharmacol:27–52Google Scholar
  21. Han M, Huang XF, Deng C (2009) Aripiprazole differentially affects mesolimbic and nigrostriatal dopaminergic transmission: implications for long-term drug efficacy and low extrapyramidal side-effects. Int J Neuropsychopharmacol 12:941–952CrossRefGoogle Scholar
  22. Harada WJ, Haycock JW, Goldstein M (1996) Regulation of L-DOPA biosynthesis by site-specific phosphorylation of tyrosine hydroxylase in AtT-20 cells expressing wild-type and serine 40-substituted enzyme. J Neurochem 67:629–635CrossRefGoogle Scholar
  23. Lerner TN, Shilyansky C, Davidson TJ, Evans KE, Beier KT, Zalocusky KA, Crow AK, Malenka RC, Luo L, Tomer R, Deisseroth K (2015) Intact-brain analyses reveal distinct information carried by SNc dopamine subcircuits. Cell 162:635–647CrossRefGoogle Scholar
  24. Li M (2016) Antipsychotic-induced sensitization and tolerance: behavioral characteristics, developmental impacts, and neurobiological mechanisms. J Psychopharmacol 30:749–770CrossRefGoogle Scholar
  25. Li X, Rosborough KM, Friedman AB, Zhu W, Roth KA (2007) Regulation of mouse brain glycogen synthase kinase-3 by atypical antipsychotics. Int J Neuropsychopharmacol 10:7–19CrossRefGoogle Scholar
  26. Lindgren N, Usiello A, Goiny M, Haycock J, Erbs E, Greengard P, Hokfelt T, Borrelli E, Fisone G (2003) Distinct roles of dopamine D2L and D2S receptor isoforms in the regulation of protein phosphorylation at presynaptic and postsynaptic sites. Proc Natl Acad Sci U S A 100:4305–4309CrossRefGoogle Scholar
  27. Long Q, Luo Q, Wang K, Bates A, Shetty AK (2017) Mash1-dependent notch signaling pathway regulates GABAergic neuron-like differentiation from bone marrow-derived mesenchymal stem cells. Aging Dis 8:301–313CrossRefGoogle Scholar
  28. Mace S, Taylor D (2009) Aripiprazole: dose-response relationship in schizophrenia and schizoaffective disorder. CNS Drugs 23:773–780CrossRefGoogle Scholar
  29. Mailman RB, Murthy V (2010) Third generation antipsychotic drugs: partial agonism or receptor functional selectivity? Curr Pharm Des 16:488–501CrossRefGoogle Scholar
  30. Moran-Gates T, Gan L, Park YS, Zhang K, Baldessarini RJ, Tarazi FI (2006) Repeated antipsychotic drug exposure in developing rats: dopamine receptor effects. Synapse 59:92–100CrossRefGoogle Scholar
  31. Morikawa H, Paladini CA (2011) Dynamic regulation of midbrain dopamine neuron activity: intrinsic, synaptic, and plasticity mechanisms. Neuroscience 198:95–111CrossRefGoogle Scholar
  32. Natesan S, Reckless GE, Nobrega JN, Fletcher PJ, Kapur S (2006) Dissociation between in vivo occupancy and functional antagonism of dopamine D2 receptors: comparing aripiprazole to other antipsychotics in animal models. Neuropsychopharmacology 31:1854–1863CrossRefGoogle Scholar
  33. Pan B, Chen J, Lian J, Huang XF, Deng C (2015) Unique effects of acute aripiprazole treatment on the dopamine D2 receptor downstream cAMP-PKA and Akt-GSK3beta signalling pathways in rats. PLoS One 10:e0132722CrossRefGoogle Scholar
  34. Pan B, Huang XF, Deng C (2016a) Aripiprazole and haloperidol activate GSK3beta-dependent signalling pathway differentially in various brain regions of rats. Int J Mol Sci 17:459CrossRefGoogle Scholar
  35. Pan B, Huang XF, Deng C (2016b) Chronic administration of aripiprazole activates GSK3beta-dependent signalling pathways, and up-regulates GABAA receptor expression and CREB1 activity in rats. Sci Rep 6:30040CrossRefGoogle Scholar
  36. Pan B, Lian J, Huang XF, Deng C (2016c) Aripiprazole increases the PKA signalling and expression of the GABAA receptor and CREB1 in the nucleus accumbens of rats. J Mol Neurosci 59:36–47CrossRefGoogle Scholar
  37. Park SW, Seo MK, Cho HY, Lee JG, Lee BJ, Seol W, Kim YH (2011) Differential effects of amisulpride and haloperidol on dopamine D2 receptor-mediated signaling in SH-SY5Y cells. Neuropharmacology 61:761–769CrossRefGoogle Scholar
  38. Paxinos G, Watson C (2005) The rat brain in stereotaxic coordinates. Elsevier Academic Press, San DiegoGoogle Scholar
  39. Reagan-Shaw S, Nihal M, Ahmad N (2008) Dose translation from animal to human studies revisited. FASEB J 22:659–661CrossRefGoogle Scholar
  40. Roskoski R Jr, Roskoski LM (1987) Activation of tyrosine hydroxylase in PC12 cells by the cyclic GMP and cyclic AMP second messenger systems. J Neurochem 48:236–242CrossRefGoogle Scholar
  41. Roth BL, Driscol J (2018) PDSP Ki database Psychoactive Drug Screening Program (PDSP). University of North Carolina at Chapel Hill and the United States National Institute of Mental HealthGoogle Scholar
  42. Seo MK, Lee CH, Cho HY, You YS, Lee BJ, Lee JG, Park SW, Kim YH (2015) Effects of antipsychotic drugs on the expression of synapse-associated proteins in the frontal cortex of rats subjected to immobilization stress. Psychiatry Res 229:968–974CrossRefGoogle Scholar
  43. Sibley DR (1999) New insights into dopaminergic receptor function using antisense and genetically altered animals. Annu Rev Pharmacol Toxicol 39:313–341CrossRefGoogle Scholar
  44. Singh KK (2013) An emerging role for Wnt and GSK3 signaling pathways in schizophrenia. Clin Genet 83:511–517CrossRefGoogle Scholar
  45. Strait KA, Kuczenski R (1986) Dopamine autoreceptor regulation of the kinetic state of striatal tyrosine hydroxylase. Mol Pharmacol 29:561–569Google Scholar
  46. Sutton LP, Rushlow WJ (2011) The effects of neuropsychiatric drugs on glycogen synthase kinase-3 signaling. Neuroscience 199:116–124CrossRefGoogle Scholar
  47. Tadori Y, Miwa T, Tottori K, Burris KD, Stark A, Mori T, Kikuchi T (2005) Aripiprazole's low intrinsic activities at human dopamine D2L and D2S receptors render it a unique antipsychotic. Eur J Pharmacol 515:10–19CrossRefGoogle Scholar
  48. Tadori Y, Kitagawa H, Forbes RA, McQuade RD, Stark A, Kikuchi T (2007) Differences in agonist/antagonist properties at human dopamine D(2) receptors between aripiprazole, bifeprunox and SDZ 208-912. Eur J Pharmacol 574:103–111CrossRefGoogle Scholar
  49. Turalba AV, Leite-Morris KA, Kaplan GB (2004) Antipsychotics regulate cyclic AMP-dependent protein kinase and phosphorylated cyclic AMP response element-binding protein in striatal and cortical brain regions in mice. Neurosci Lett 357:53–57CrossRefGoogle Scholar
  50. Wadenberg M-LG (2007) Bifeprunox: a novel antipsychotic agent with partial agonist properties at dopamine D2 and serotonin 5-HT1A receptors. Future Neurol 2:153–165CrossRefGoogle Scholar
  51. Wang B, Zhang Y, Dong H, Gong S, Wei B, Luo M, Wang H, Wu X, Liu W, Xu X, Zheng Y, Sun M (2018) Loss of Tctn3 causes neuronal apoptosis and neural tube defects in mice. Cell Death Dis 9:520CrossRefGoogle Scholar
  52. Wolf ME, Roth RH (1990) Autoreceptor regulation of dopamine synthesis. Ann N Y Acad Sci 604:323–343CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The Key Laboratory of Syndrome Differentiation and Treatment of Gastric Cancer of the State Administration of Traditional Chinese MedicineYangzhou University Medical CollegeYangzhouChina
  2. 2.Department of PharmacyYangzhou University Medical CollegeYangzhouChina
  3. 3.School of MedicineUniversity of WollongongWollongongAustralia
  4. 4.Antipsychotic Research Laboratory, Illawarra Health and Medical Research InstituteUniversity of WollongongWollongongAustralia

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