Advertisement

Molecular Biology Reports

, Volume 39, Issue 2, pp 2003–2010 | Cite as

Use of siRNA in knocking down of dopamine receptors, a possible therapeutic option in neuropsychiatric disorders

  • Mohammad-Reza Noori-DaloiiEmail author
  • Majid Mojarrad
  • Ali Rashidi-nezhad
  • Majid Kheirollahi
  • Ali Shahbazi
  • Mehdi Khaksari
  • Asghar Korzebor
  • Ali Goodarzi
  • Maryam Ebrahimi
  • Ali Reza Noori-Daloii
Article

Abstract

Heightened dopaminergic activity has been shown to be implicated in some major neuropsychiatric disorders such as schizophrenia. Use of dopaminergic antagonists was limited by some serious side effects related to unspecific blocking of dopamine receptors. Thus a target specific dopamine receptor gene silencing method such as using small interfering RNA (siRNA) might be useful. In this study recombinant plasmids expressing siRNA against dopamine receptors (D1-D5DRs) were produced, and their efficiency in knocking down of receptors in were assessed in rat neuroblastoma cell line (B65), using Real-time PCR method. Furthermore, D2DR siRNA expressing plasmid was injected into the rat nucleus accumbens bilaterally to investigate whether it can prevent the hyperactivity induced by apomorphine. Locomotion was measured in 10 min intervals, 50 min before and 60 min after apomorphine injection (0.5 mg/kg, S.C). Our results indicated that the mRNA level of dopamine receptors were reduced between 25 and 75% in B65 cells treated with the plasmids in vitro. In behavioral tests, locomotion was lower at least in the second 10 min after apomorphine injection in rats treated with plasmid expressing D2DR siRNA compare to control group [F (4,24) = 2.77, (P < 0.05)]. The spontaneous activity of treated rats was normal. In conclusion, dopamine receptors can be downregulated by use of siRNA expressing plasmids in nucleus accumbens. Although our work may have some possible clinical applications; the potentially therapeutic application of siRNA in knocking down of dopamine receptors needs further studies.

Keywords

siRNA Dopamine receptors Schizophrenia D2DR Nucleus accumbens Locomotion 

Notes

Acknowledgments

This research has been supported by the Iran National Science Foundation (INSF), grant no. 83088.

References

  1. 1.
    Vijayraghavan S, Wang M, Birnbaum SG, Williams GV, Arnsten AF (2007) Inverted-U dopamine D1 receptor actions on prefrontal neurons engaged in working memory. Nat Neurosci 10:376–384PubMedCrossRefGoogle Scholar
  2. 2.
    Floresco SB, Tse MT, Ghods-Sharifi S (2008) Dopaminergic and glutamatergic regulation of effort- and delay-based decision making. Neuropsychopharmacology 33:1966–1979PubMedCrossRefGoogle Scholar
  3. 3.
    Nieoullon A (2002) Dopamine and the regulation of cognition and attention. Prog Neurobiol 67:53–83PubMedCrossRefGoogle Scholar
  4. 4.
    Bernheimer H, Birkmayer W, Hornykiewicz O, Jellinger K, Seitelberger F (1973) Brain dopamine and the syndromes of Parkinson and Huntington. Clinical, morphological and neurochemical correlations. J Neurol Sci 20:415–455PubMedCrossRefGoogle Scholar
  5. 5.
    Grace AA, Floresco SB, Goto Y, Lodge DJ (2007) Regulation of firing of dopaminergic neurons and control of goal-directed behaviors. Trends Neurosci 30:220–227PubMedCrossRefGoogle Scholar
  6. 6.
    Rosenkranz JA, Grace AA (1999) Modulation of basolateral amygdala neuronal firing and afferent drive by dopamine receptor activation in vivo. J Neurosci 19:11027–11039PubMedGoogle Scholar
  7. 7.
    Arnsten AF (2009) Toward a new understanding of attention-deficit hyperactivity disorder pathophysiology: an important role for prefrontal cortex dysfunction. CNS Drugs 23(Suppl 1):33–41PubMedCrossRefGoogle Scholar
  8. 8.
    Bonci A, Bernardi G, Grillner P, Mercuri NB (2003) The dopamine-containing neuron: maestro or simple musician in the orchestra of addiction? Trends Pharmacol Sci 24:172–177PubMedCrossRefGoogle Scholar
  9. 9.
    Greene JG (2006) Gene expression profiles of brain dopamine neurons and relevance to neuropsychiatric disease. J Physiol 575:411–416PubMedCrossRefGoogle Scholar
  10. 10.
    Kebabian JW, Petzold GL, Greengard P (1972) Dopamine-sensitive adenylate cyclase in caudate nucleus of rat brain, and its similarity to the “dopamine receptor”. Proc Natl Acad Sci USA 69:2145–2149PubMedCrossRefGoogle Scholar
  11. 11.
    Seeman P, Chau-Wong M, Tedesco J, Wong K (1975) Brain receptors for antipsychotic drugs and dopamine: direct binding assays. Proc Natl Acad Sci USA 72:4376–4380PubMedCrossRefGoogle Scholar
  12. 12.
    Werkman TR, Glennon JC, Wadman WJ, McCreary AC (2006) Dopamine receptor pharmacology: interactions with serotonin receptors and significance for the aetiology and treatment of schizophrenia. CNS Neurol Disord Drug Targets 5:3–23PubMedCrossRefGoogle Scholar
  13. 13.
    Strange PG (1993) Dopamine receptors: structure and function. Prog Brain Res 99:167–179PubMedCrossRefGoogle Scholar
  14. 14.
    Vallone D, Picetti R, Borrelli E (2000) Structure and function of dopamine receptors. Neurosci Biobehav Rev 24:125–132PubMedCrossRefGoogle Scholar
  15. 15.
    Birkmayer W, Hornykiewicz O (1961) The L-3,4-dioxyphenylalanine (DOPA)-effect in Parkinson-akinesia. Wien Klin Wochenschr 73:787–788PubMedGoogle Scholar
  16. 16.
    Carlsson A, Lindqvist M (1963) Effect of chlorpromazine or haloperidol on formation of 3methoxytyramine and normetanephrine in mouse brain. Acta Pharmacol Toxicol (Copenh) 20:140–144CrossRefGoogle Scholar
  17. 17.
    Iversen SD, Iversen LL (2007) Dopamine: 50 years in perspective. Trends Neurosci 30:188–193PubMedCrossRefGoogle Scholar
  18. 18.
    Sah DW (2006) Therapeutic potential of RNA interference for neurological disorders. Life Sci 79:1773–1780PubMedCrossRefGoogle Scholar
  19. 19.
    Lieberman JA, Stroup TS, McEvoy JP, Swartz MS, Rosenheck RA et al (2005) Effectiveness of antipsychotic drugs in patients with chronic schizophrenia. N Engl J Med 353:1209–1223PubMedCrossRefGoogle Scholar
  20. 20.
    Remington G, Kapur S (2010) Antipsychotic dosing: how much but also how often? Schizophr Bull 36:900–903PubMedCrossRefGoogle Scholar
  21. 21.
    Schultz SH, North SW, Shields CG (2007) Schizophrenia: a review. Am Fam Physician 75:1821–1829PubMedGoogle Scholar
  22. 22.
    Aagaard L, Rossi JJ (2007) RNAi therapeutics: principles, prospects and challenges. Adv Drug Deliv Rev 59:75–86PubMedCrossRefGoogle Scholar
  23. 23.
    Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE et al (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811PubMedCrossRefGoogle Scholar
  24. 24.
    Kim DH, Rossi JJ (2007) Strategies for silencing human disease using RNA interference. Nat Rev Genet 8:173–184PubMedCrossRefGoogle Scholar
  25. 25.
    Takahashi Y, Nishikawa M, Takakura Y (2009) Nonviral vector-mediated RNA interference: its gene silencing characteristics and important factors to achieve RNAi-based gene therapy. Adv Drug Deliv Rev 61:760–766PubMedCrossRefGoogle Scholar
  26. 26.
    Thakker DR, Hoyer D, Cryan JF (2006) Interfering with the brain: use of RNA interference for understanding the pathophysiology of psychiatric and neurological disorders. Pharmacol Ther 109:413–438PubMedCrossRefGoogle Scholar
  27. 27.
    Makimura H, Mizuno TM, Mastaitis JW, Agami R, Mobbs CV (2002) Reducing hypothalamic AGRP by RNA interference increases metabolic rate and decreases body weight without influencing food intake. BMC Neurosci 3:18PubMedCrossRefGoogle Scholar
  28. 28.
    Paxinos G, Watson C (1998) The rat brain in stereotaxic coordinates. Academic Press, New YorkGoogle Scholar
  29. 29.
    Wadhwa R, Kaul SC, Miyagishi M, Taira K (2004) Know-how of RNA interference and its applications in research and therapy. Mutat Res 567:71–84PubMedCrossRefGoogle Scholar
  30. 30.
    Backman C, Zhang Y, Hoffer BJ, Tomac AC (2003) Short interfering RNAs (siRNAs) for reducing dopaminergic phenotypic markers. J Neurosci Methods 131:51–56PubMedCrossRefGoogle Scholar
  31. 31.
    Abi-Dargham A, Rodenhiser J, Printz D, Zea-Ponce Y, Gil R et al (2000) Increased baseline occupancy of D2 receptors by dopamine in schizophrenia. Proc Natl Acad Sci USA 97:8104–8109PubMedCrossRefGoogle Scholar
  32. 32.
    Barr AM, Powell SB, Markou A, Geyer MA (2006) Iloperidone reduces sensorimotor gating deficits in pharmacological models, but not a developmental model, of disrupted prepulse inhibition in rats. Neuropharmacology 51:457–465PubMedCrossRefGoogle Scholar
  33. 33.
    Breysse N, Risterucci C, Amalric M (2002) D1 and D2 dopamine receptors contribute to the locomotor response induced by Group II mGluRs activation in the rat nucleus accumbens. Pharmacol Biochem Behav 73:347–357PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Mohammad-Reza Noori-Daloii
    • 1
    Email author
  • Majid Mojarrad
    • 1
  • Ali Rashidi-nezhad
    • 1
  • Majid Kheirollahi
    • 1
  • Ali Shahbazi
    • 2
    • 3
  • Mehdi Khaksari
    • 2
  • Asghar Korzebor
    • 2
  • Ali Goodarzi
    • 3
  • Maryam Ebrahimi
    • 1
  • Ali Reza Noori-Daloii
    • 1
  1. 1.Department of Medical Genetics, Faculty of MedicineTehran University of Medical SciencesTehranIran
  2. 2.Physiology Research Center (PRC)Tehran University of Medical SciencesTehranIran
  3. 3.Institute for Cognitive Science Studies (ICSS)TehranIran

Personalised recommendations