Alcoholism and GRM7

Jennifer 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 state

Psychological factors

Social and cultural factors

Age, sex

Genetics (including GRM7)

Alcoholism – Risk Factors

Mayo Foundation for Medical Education and Research (MFMER), 2008

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Vadasz et al. 2007

Discovered link between

alcoholism 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), 2008

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GRM7

ANF domain: extra-cellular ligand binding

NCDG3: stable linker composed of disulphide bridges

7tm_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

RNAi

C. elegans – mgl-1

post-embryonic slow-growth rate

D. melanogaster – mGluRA

neurophysiological 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 - Similarities

PICK1

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

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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 - Databases

AmiGO: 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/