Home LiteratureArticle Details
PMID: 12692303 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Gamma-aminobutyric acid B receptor 1 mediates behavior-impairing actions of alcohol in Drosophila: adult RNA interference and pharmacological evidence.

Dzitoyeva S, Dimitrijevic N, Manev H

Abstract

In addition to their physiological function, metabotropic receptors for neurotransmitter gamma-aminobutyric acid (GABA), the GABA(B) receptors, may play a role in the behavioral actions of addictive compounds. Recently, GABA(B) receptors were cloned in fruit flies (Drosophila melanogaster), indicating that the advantages of this experimental model could be applied to GABA(B) receptor research. RNA interference (RNAi) is an endogenous process triggered by double-stranded RNA and is being used as a tool for functional gene silencing and functional genomics. Here we show how cell-nonautonomous RNAi can be induced in adult fruit flies to silence a subtype of GABA(B) receptors, GABA(B)R1, and how RNAi combined with pharmacobehavioral techniques (including intraabdominal injections of active compounds and a computer-assisted quantification of behavior) can be used to functionally characterize these receptors. We observed that injection of double-stranded RNA complementary to GABA(B)R1 into adult Drosophila selectively destroys GABA(B)R1 mRNA and attenuates the behavioral actions of the GABA(B) agonist, 3-aminopropyl-(methyl)phosphinic acid. Moreover, both GABA(B)R1 RNAi and the GABA(B) antagonist CGP 54626 reduced the behavior-impairing effects of ethanol, suggesting a putative role for the Drosophila GABA(B) receptors in alcohol's mechanism of action. The Drosophila model we have developed can be used for further in vivo functional characterization of GABA(B) receptor subunits and their involvement in the molecular and systemic actions of addictive substances.

MeSH Terms
Animals Base Sequence Behavior, Animal/drug effects DNA Primers Drosophila melanogaster Ethanol/pharmacology Gene Silencing In Vitro Techniques RNA Interference RNA, Messenger/genetics Receptors, GABA-B/genetics,physiology
Chemicals
DNA Primers RNA, Messenger Receptors, GABA-B Ethanol
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Dzitoyeva Svetlana
Department of Psychiatry, Psychiatric Institute, University of Illinois, Chicago, IL 60612, USA.
Dimitrijevic Nikola
Manev Hari
References (32)
32 references, click to expand
  1. GABA(A) receptors as molecular sites of ethanol action. Direct or indirect actions?
    Curr Top Med Chem. 2002 Aug;2(8):869-85 PMID: 12171577
  2. A single subunit (GB2) is required for G-protein activation by the heterodimeric GABA(B) receptor.
    J Biol Chem. 2002 Feb 1;277(5):3236-41 PMID: 11711539
  3. A new alcohol antagonist: phaclofen.
    Life Sci. 1989;45(19):1771-9 PMID: 2556616
  4. Human GABA(B)R genomic structure: evidence for splice variants in GABA(B)R1 but not GABA(B)R2.
    Gene. 2001 Oct 31;278(1-2):63-79 PMID: 11707323
  5. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.
    Nature. 1998 Feb 19;391(6669):806-11 PMID: 9486653
  6. RNA interference.
    Nature. 2002 Jul 11;418(6894):244-51 PMID: 12110901
  7. Long-lasting potentiation of GABAergic synapses in dopamine neurons after a single in vivo ethanol exposure.
    J Neurosci. 2002 Mar 15;22(6):2074-82 PMID: 11896147
  8. Dopamine modulates acute responses to cocaine, nicotine and ethanol in Drosophila.
    Curr Biol. 2000 Feb 24;10(4):187-94 PMID: 10704411
  9. Heritable gene silencing in Drosophila using double-stranded RNA.
    Nat Biotechnol. 2000 Aug;18(8):896-8 PMID: 10932163
  10. Epilepsy, hyperalgesia, impaired memory, and loss of pre- and postsynaptic GABA(B) responses in mice lacking GABA(B(1)).
    Neuron. 2001 Jul 19;31(1):47-58 PMID: 11498050
  11. Epileptogenesis and enhanced prepulse inhibition in GABA(B1)-deficient mice.
    Mol Cell Neurosci. 2001 Jun;17(6):1059-70 PMID: 11414794
  12. Combinatorial RNAi: a method for evaluating the functions of gene families in Drosophila.
    Trends Neurosci. 2002 Feb;25(2):71-4 PMID: 11814553
  13. Proconvulsive effect of the GABA(B) receptor antagonist, SCH 50911, in rats undergoing ethanol withdrawal syndrome.
    Eur J Pharmacol. 2002 Jun 12;445(3):195-9 PMID: 12079684
  14. Baclofen efficacy in reducing alcohol craving and intake: a preliminary double-blind randomized controlled study.
    Alcohol Alcohol. 2002 Sep-Oct;37(5):504-8 PMID: 12217947
  15. Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA.
    BMC Biotechnol. 2003 Jan 20;3:1 PMID: 12546706
  16. International Union of Pharmacology. XXXIII. Mammalian gamma-aminobutyric acid(B) receptors: structure and function.
    Pharmacol Rev. 2002 Jun;54(2):247-64 PMID: 12037141
  17. Ethanol intoxication in Drosophila: Genetic and pharmacological evidence for regulation by the cAMP signaling pathway.
    Cell. 1998 Jun 12;93(6):997-1007 PMID: 9635429
  18. Functional ethanol tolerance in Drosophila.
    Neuron. 2000 Oct;28(1):261-71 PMID: 11086999
  19. GABA(B) receptors play a role in the development of tolerance to ethanol in mice.
    Psychopharmacology (Berl). 2001 Feb;153(4):415-24 PMID: 11243488
  20. G-protein-coupled inwardly rectifying potassium channels are targets of alcohol action.
    Nat Neurosci. 1999 Dec;2(12):1084-90 PMID: 10570485
  21. Increased selection response in larger populations. II. Selection for ethanol vapor resistance in Drosophila melanogaster at two population sizes.
    Genetics. 1990 Jul;125(3):585-97 PMID: 2116359
  22. Functional genomic analysis of C. elegans chromosome I by systematic RNA interference.
    Nature. 2000 Nov 16;408(6810):325-30 PMID: 11099033
  23. Novel genomic cDNA hybrids produce effective RNA interference in adult Drosophila.
    Neuron. 2002 Jan 17;33(2):177-84 PMID: 11804566
  24. Ancient pathways programmed by small RNAs.
    Science. 2002 May 17;296(5571):1265-9 PMID: 12016303
  25. Mouse cerebellar GABAB participation in the expression of acute ethanol-induced ataxia and in its modulation by the cerebellar adenosinergic A1 system.
    Brain Res Bull. 1996;41(1):53-9 PMID: 8883916
  26. Cloning and functional expression of GABA(B) receptors from Drosophila.
    Eur J Neurosci. 2001 Feb;13(3):477-86 PMID: 11168554
  27. Two-color GFP imaging demonstrates cell-autonomy of GAL4-driven RNA interference in Drosophila.
    Genesis. 2002 Sep-Oct;34(1-2):170-3 PMID: 12324976
  28. Intra-abdominal injection of double-stranded RNA into anesthetized adult Drosophila triggers RNA interference in the central nervous system.
    Mol Psychiatry. 2001 Nov;6(6):665-70 PMID: 11673794
  29. Hemolin gene silencing by ds-RNA injected into Cecropia pupae is lethal to next generation embryos.
    Insect Mol Biol. 2002 Jun;11(3):267-71 PMID: 12000646
  30. Efficient and heritable functional knock-out of an adult phenotype in Drosophila using a GAL4-driven hairpin RNA incorporating a heterologous spacer.
    Nucleic Acids Res. 2001 Jun 15;29(12):E55-5 PMID: 11410678
  31. Role for a bidentate ribonuclease in the initiation step of RNA interference.
    Nature. 2001 Jan 18;409(6818):363-6 PMID: 11201747
  32. A GABAB receptor on an identified insect motor neurone
    J Exp Biol. 1995;198(Pt 4):889-94 PMID: 9318675
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2003-04-29
Epub
2003-00-11
Pages
5485-90
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC154371
Subset
IM
Grants
NIDA NIH HHS · R03DA14811 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]