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Alcoholism and GRM7Jennifer Wagner
Image from http://web.centre.edu/smart/help.html
Due to an imbalance inneurotransmitter levels leading to…1) Cravings2) Loss of control3) Withdrawal4) Tolerance
Alcoholism – The Disease
Mayo Foundation for Medical Education and Research (MFMER), 2008
Image from http://www.blog.joelx.com/wp-content/uploads/2008/04/alcoholism-scotch.gifVideo
Emotional statePsychological factors
Social and cultural factorsAge, sex
Genetics (including GRM7)
Alcoholism – Risk Factors
Mayo Foundation for Medical Education and Research (MFMER), 2008
Images from http://www.blog.joelx.com/wp-content/uploads/2008/04/alcoholism-scotch.gif, http://www.cusmibio.unimi.it/documenti/website_new/en/introduction/index2.htm
Vadasz et al. 2007
Discovered link betweenalcoholism and GRM7
Lowered GRM7 expression correlated to increased preference for alcohol
Image from http://www.jax.org/strain/000664.html
Affects 1 in 12 Americans
Leads to damaged relationships, financial problems, and unemployment, as well as severe health problems
Understanding the genetic causes may lead to development of better prevention as well as new drug treatments
Alcoholism – Why study the genes involved?
Mayo Foundation for Medical Education and Research (MFMER), 2008Image from http://www.blog.joelx.com/wp-content/uploads/2008/04/alcoholism-scotch.gif
GRM7
ANF domain: extra-cellular ligand bindingNCDG3: stable linker composed of disulphide
bridges7tm_3: transmembrane region, signal transducer
G-protein coupled metabotropic glutamate receptor:
SMART database
Gene OntologyBiological Processes:
synaptic transmission, conditioned taste aversion, learning and memory, behavioral fear response, transmission of nerve impulse, multicellular organismal response to stress
Cellular Components:integral to plasma membrane, postsynaptic membrane, presynaptic active zone
Molecular Functions:glutamate binding, calmodulin binding, calcium channel regulator, PDZ domain binding
AmiGO database
Expression Patterns
Oscar-Berman and Marinkovic, 2003.
MicroArray dataSamples from post-mortem humans averaging 70 yrs
GRM7 expression shown to be lower in males
GEO database; Rinn, et al., 2004
GRM7 Homologs
Entrez Protein database; Anisimova, et al., 2006; Castresana, 2000; Chevenet, et al., 2008; Dereeper, et al., 2008; Guindon, et al., 2003; Notredame, et al., 2000
GRM7 HomologsHomo sapiens
GRM7
Mus musculusGrm7 – 99% identity
Rattus norvegicusGrm7 – 99% identity
Drosophila melanogastermGluRA – 47% identity
Caenorhabditis elegans mgl-1 – 38% identity
Entrez Protein database; Altschul, et al., 2005; Altschul, et al., 1997
Images from Pfam database
RNAiC. elegans – mgl-1
post-embryonic slow-growth rate
D. melanogaster – mGluRAneurophysiological defects in presynaptic excitability and synaptic morphology
Wormbase; Kamath, et al., 2003; Flybase; Bogdanik, 2004 Images from http://www.cafepress.com/celegans, http://flybase.org
Chemical Genetics
AMN082 – a selective GRM7 agonist: caused decreased alcohol preference in mice
MMPIP – a selective GRM7 antagonist: potential for use in future studies
Tocris bioscience, 2009; Salling, et al., 2008; Suzuki, et al., 2007; Flor, et al., 2008; Mitsukawa, et al., 2005
AMN082 MMPIP
Protein Interactions
STRING database
Text-mining evidence
Experimental evidence
Protein Interactions - SimilaritiesPICK1
PKCA
CAM3
MARKSL1
SYN47Vesl-2
SMART database
Future Directions
IP followed by mass spectrometry to confirm predicted protein interactions
Exploring protein interactions
MicroArray studies in post-mortem samples from human alcoholics vs. non-alcoholics
Image from http://www.bgx.org.uk/microarray.jpg
Conclusions
Lowered expression of GRM7 correlates to increased susceptibility to alcoholism
GRM7 may not be a good target for therapeutic intervention in alcoholism
However, learning more about it may lead to useful treatments
Image from http://www.blog.joelx.com/wp-content/uploads/2008/04/alcoholism-scotch.gif
Questions?
References -LiteratureAltschul, S.F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D. J. (1997). Gapped
BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25:3389-3402.
Altschul, S. F., Wootton, J. C., Gertz, E. M., Agarwala, R., Morgulis, A., Schäffer, A. A., and Yu, Y. K. (2005). Protein database searches using compositionally adjusted substitution matrices. FEBS J., 272:5101-5109.
Bogdanik, L., Mohrmann, R., Ramaekers, A., Bockaert, J., Grau, Y., Broadie, K., Parmentier, M.L. (2004). The Drosophila metabotropic glutamate receptor DmGluRA regulates activity-dependent synaptic facilitation and fine synaptic morphology. J. Neurosci. 24(41): 9105--9116.
Castresana J. (2000). Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Mol Biol Evol. 17(4):540-52.
Chevenet F., Brun C., Banuls AL., Jacq B., Chisten R. (2008) TreeDyn: towards dynamic graphics and annotations for analyses of trees. BMC Bioinformatics. 7:439.
Dereeper A.*, Guignon V.*, Blanc G., Audic S., Buffet S., Chevenet F., Dufayard J.F., Guindon S., Lefort V., Lescot M., Claverie J.M., Gascuel O. (2008). Phylogeny.fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res. 36(Web Server issue):W465-9. *joint first authors
Flor, P.J., Vassout, A., Fendt, M., Mitsukawa, K., Jacobson, L., Yamamoto, R., Nozulak, J., Ofner, S., Pescott, O., Lukic, S., Stoehr, N., Urwyler, S., Kuhn, R., Herrling, P., McAllister, K., van der Putten, H., and Cryan, J.F. (2005). AMN082, the first selective mGluR7 agonist: activation of receptor signaling via an allosteric site in the transmembrane domain modulates stress parameters in vivo. [Abstract]. Neuropharmacology, 49 (Suppl. 1):244. doi: 10.1016/j.neuropharm.2005.06.013
Guindon S., Gascuel O. (2003). A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol. 52(5):696-704.
Kamath, R.S., Fraser, A.G., Dong, Y., Poulin, G., Durbin, R., Gotta, M., Kanapin, A., Le Bot, N., Moreno, S., Sohrmann, M., Welchman, D.P., Zipperlen, P., Ahringer, J. (2003). Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature, 421(6920):231-7. doi:10.1038/nature01278
Mayo Foundation for Medical Education and Research (MFMER). (2008, May 8). Alcoholism. Retrieved February 2, 2009, from http://www.mayoclinic.com/print/alcoholism/DS00340/METHOD=print&DSECTION=all
References -LiteratureMitsukawa, K., Yamamoto, R., Ofner, S., Nozulak, J., Pescott, O., Lukic, S., Stoehr, N., Mombereau,
C., Kuhn, R., McAllister, K., van der Putten, H., Cryan, J.F., and Flor, P.J. (2005). A selective metabotropic glutamate receptor 7 agonist: Activation of receptor signaling via an allosteric site modulates stress parameters in vivo. Proc. Natl. Acad. Sci. USA ,102(51):18712. doi: 10.1073/pnas.0508063102.
Notredame C., Higgins DG., Heringa J. (2000). T-Coffee: A novel method for fast and accurate multiple sequence alignment. J
Oscar-Berman, M. and Marinkovic, K. (2003). Alcoholism and the brain: An overview. Alcohol Research and Health, 27(2), 125. Retrieved February 2, 2009, from: http://web.ebscohost.com.ezproxy.library.wisc.edu/ehost/detail?vid=1&hid=7&sid=07de3f40-7d7a-4f64-ad4a-fb9e6825d1ad%40sessionmgr2&bdata=JnNpdGU9ZWhvc3QtbGl2ZQ%3d%3d#db=rzh&AN=2004159543
Rinn, J.L., Rozowsky, J.S., Laurenzi, I.J., Petersen, P., Zon, K., Zhong, W., Gerstein, M., Snyder, M.P. (2004). Sex Specific Transcription in Human Hypothalamus. Retrieved April 22, 2009 from http://www.ncbi.nlm.nih.gov/geo/gds/profileGraph.cgi?&dataset=595j46FBzmACVZ2lmXkl28-&dataset=dfcjbcRKmkHM7-ajk8jjbff$&gmin=24.600000&gmax=750.500000&absc=19194p2p1p1p1p3p1p3p3p3&gds=564&idref=207548_at&annot=GRM7
Salling, M.C., Faccidomo, S., Hodge, C.W. (2008). Nonselective suppression of operant ethanol and sucrose self-administration by the mGluR7 positive allosteric modulator AMN082. Pharmacology Biochemistry and Behavior, 91(1):14. doi: 10.1016/j.pbb.2008.06.006
Suzuki, G., Tsukamoto, N., Fushiki, H., Kawagishi, A., Nakamura, M., Kurihara, H., Mitsuya, M., Ohkubo, M., and Ohta, H. (2007). In Vitro Pharmacological Characterization of Novel Isoxazolopyridone Derivatives as Allosteric Metabotropic Glutamate Receptor 7 Antagonists . J Pharmacol Exp Ther, 323(1):147. doi: 10.1124/jpet.107.124701
Tocris Bioscience. (2009). AMN082 dihydrochloride. Retrieved May 2, 2009, from http://www.tocris.com/dispprod.php?ItemId=145226.
Tocris Bioscience. (2009). MMPIP hydrochloride. Retrieved May 3, 2009, from http://www.tocris.com/dispprod.php?ItemId=5430.
Vadasz, C., Saito, M., Gyetvai, B. M., Oros, M., Szakall, I., Kovacs, K. M., Prasad, V. V. T. S., Toth, R. (2007). Glutamate receptor metabotropic 7 is cis-regulated in the mouse brain and modulates alcohol drinking. Genomics, 90(6):690. doi:10.1016/j.ygeno.2007.08.006
References - DatabasesAmiGO:
http://amigo.geneontology.org/cgi-bin/amigo/go.cgi
Entrez Protein: http://www.ncbi.nlm.nih.gov/protein/Flybase: http://flybase.org/GEO: http://www.ncbi.nlm.nih.gov/geo/Pfam: http://pfam.sanger.ac.uk/SMART: http://smart.embl-heidelberg.de/STRING: http://string.embl.de/Wormbase: http://www.wormbase.org/