Genetic association of nucleus accumbens 5-hydroxyindoleacetic acid level and alcohol preference drinking in a quasi-congenic male mice: Potential modulation by Grm7 gene polymorphism.

Objective: To test the hypothesis that predisposition to high alcohol drinking behavior is genetically associated with hypoactive serotonergic function in the Nucleus Accumbens (NAc). Method: Alcohol avoiding C5A3 and alcohol preferring I5B25A mice of the Quasi-congenic Recombinant QTL Introgression (RQI) mouse strains were subjected to in vivo microdialysis in the NAc. Neurotransmitter and metabolite contents were analyzed by HPLC and samples were collected in three phases: Baseline, Control, and Alcohol. Samples were collected with 20 min intervals. Results: Between-strain differences restricted to small chromosome segments significantly affected both alcohol preference drinking and NAc 5-HIAA levels [F1, 13 = 5.569 p=.035 (General Linear Model Repeated Measures ANOVA and Tests of Between-Subjects Effects)]. Whole genome biallelic DNA marker genotyping allowed the identification of 16 differential microsatellite markers associated with low 5-HIAA levels and excessive alcohol drinking. Chromosome 6 markers were linked to Grm7 (51.19 centimorgan), a reported candidate gene for modulation of addiction. The results are consistent with earlier reports of association of low 5-HIAA and high alcohol consumption in rats and primates, including Homo sapiens. Conclusion: Low NAc 5-HIAA and high alcohol consumption are genetically associated in a quasi-congenic mouse model carrying variants of the Grm7 gene. We propose that constitutional polymorphism in Grm7 may modulate CRF neuron activity via altered mGluR7 expression thus targeting CRF pathways to substance use circuits. This raises the possibility of modulation of DRN 5-HT neurons leading to hypo- or hyper-serotonergic condition in NAc and higher or lower alcohol preference drinking.

Cocaine-Induced Sensitization is Linked to Distal Chromosome 6 Region in Congenic Mouse Model.

OBJECTIVE: Previously we mapped QTL Eac2 to mouse Chr6 and identified the first gene (Grm7) as accounting for alcohol consumption in a mammalian model. Despite the central role of glutamate receptors in addiction, the effects of Grm7 gene variants are not well known. Here we test the hypothesis that genetic variation of the distal mouse Chr6 Eac2 region, location of Grm7, controls cocaine-induced locomotor sensitization. METHOD: C57BL/6By background and B6.C6.327.54 congenic mice were subjected to whole-genome SNP genotyping. Isogeneic (C57BL/6ByXB6.C6.327.54)F2 mice homozygous for SNPs in the BALB/c-type Eac2 region were selected to create a subcongenic strain (B6By.C6.108-120). In a 2-strain x 2-sex 2-treatment factorial design (n = 6-10) C57BL/6By and B6By.C6.108-120 mice received repeated daily cocaine or saline intraperitoneal injections, and locomotor activity was recorded for 90 minutes immediately after injection. RESULTS: C57BL/6By females with the G/G genotype of SNP rs3723352 of Grm7 responded to cocaine with significantly higher activity and greater cocaine-induced sensitization than those with the BALB/cJ-type T/T genotype in the congenic strain. CONCLUSION: The results are consistent with a large body of accumulated mechanistic evidence for a role of the mGlu7 receptor in the control of neurobiological responses to cocaine, and are consistent with the hypotheses that (1) natural variants of the Grm7 gene show pleiotropy and can modulate cocaine-induced behaviors in addition to alcohol consumption, (2) interactions between mGluR7 expression, estrogen receptors, and estradiol may explain phenotypic variation in females. Heritable variation of GRM7 may affect vulnerability to substance abuse in women.

mGluR7 genetics and alcohol: intersection yields clues for addiction.

Development of addiction to alcohol or other substances can be attributed in part to exposure-dependent modifications at synaptic efficacy leading to an organism which functions at an altered homeostatic setpoint. Genetic factors may also influence setpoints and the stability of the homeostatic system of an organism. Quantitative genetic analysis of voluntary alcohol drinking, and mapping of the involved genes in the quasi-congenic Recombinant QTL Introgression strain system, identified Eac2 as a Quantitative Trait Locus (QTL) on mouse chromosome 6 which explained 18% of the variance with an effect size of 2.09 g/kg/day alcohol consumption, and Grm7 as a quantitative trait gene underlying Eac2 [Vadasz et al. in Neurochem Res 32:1099-1112, 100, Genomics 90:690-702, 102]. In earlier studies, the product of Grm7 mGluR7, a G protein-coupled receptor, has been implicated in stress systems [Mitsukawa et al. in Proc Natl Acad Sci USA 102:18712-18717, 63], anxiety-like behaviors [Cryan et al. in Eur J Neurosci 17:2409-2417, 14], memory [Holscher et al. in Learn Mem 12:450-455, 26], and psychiatric disorders (e.g., [Mick et al. in Am J Med Genet B Neuropsychiatr Genet 147B:1412-1418, 61; Ohtsuki et al. in Schizophr Res 101:9-16, 72; Pergadia et al. in Paper presented at the 38th Annual Meeting of the Behavior Genetics Association, Louisville, Kentucky, USA, 76]. Here, in experiments with mice, we show that (1) Grm7 knockout mice express increased alcohol consumption, (2) sub-congenic, and congenic mice carrying a Grm7 variant characterized by higher Grm7 mRNA drink less alcohol, and show a tendency for higher circadian dark phase motor activity in a wheel running paradigm, respectively, and (3) there are significant genetic differences in Grm7 mRNA abundance in the mouse brain between congenic and background mice identifying brain areas whose function is implicated in addiction related processes. We hypothesize that metabotropic glutamate receptors may function as regulators of homeostasis, and Grm7 (mGluR7) is involved in multiple processes (including stress, circadian activity, reward control, memory, etc.) which interact with substance use and the development of addiction. In conclusion, we suggest that mGluR7 is a significant new therapeutic target in addiction and related neurobehavioral disorders.

Glutamate receptor metabotropic 7 is cis-regulated in the mouse brain and modulates alcohol drinking.

Alcoholism is a heritable disease that afflicts about 8% of the adult population. Its development and symptoms, such as craving, loss of control, physical dependence, and tolerance, have been linked to changes in mesolimbic, mesocortical neurotransmitter systems utilizing biogenic amines, GABA, and glutamate. Identification of genes predisposing to alcoholism, or to alcohol-related behaviors in animal models, has been elusive because of variable interactions of multiple genes with relatively small individual effect size and sensitivity of the predisposing genotype to lifestyle and environmental factors. Here, using near-isogenic advanced animal models with reduced genetic background interactions, we integrate gene mapping and gene mRNA expression data in segregating and congenic mice and identify glutamate receptor metabotropic 7 (Grm7) as a cis-regulated gene for alcohol consumption. Traditionally, the mesoaccumbal dopamine reward hypothesis of addiction and the role of the ionotropic glutamate receptors have been emphasized. Our results lend support to an emerging direction of research on the role of metabotropic glutamate receptors in alcoholism and drug addiction. These data suggest for the first time that Grm7 is a risk factor for alcohol drinking and a new target in addiction therapy.

Mapping of QTLs for oral alcohol self-administration in B6.C and B6.I quasi-congenic RQI strains.

One strategy to identify neurochemical pathways of addiction is to map the relevant genes. In the present study we used 43 B6.C and 35 B6.I inbred RQI mouse strains, carrying <3% donor genome on C57BL/6ByJ background, for gene mapping. The strains were phenotyped for consumption of alcohol (12% v/v) in a two-bottle-choice paradigm, and genotyped for 396 microsatellite markers. The current mapping study extends our earlier experiment scanning five mouse chromosomes (Vadasz et al. (2000) Scanning of five chromosomes for alcohol consumption loci. Alcohol 22:25-34) to a whole-genome study, and discusses the differences and limitations. Data were analyzed with composite interval (CIM) and multiple interval (MIM) QTL mapping methods. CIM of B6.C strains detected significant QTLs on chrs. 6 and 12. A suggestive, but not significant, locus was detected in the B6.I strains on chr. 12. The best MIM model for B6.C strains confirmed one QTL on chr. 6 and one QTL on chr. 12, while the MIM model for the B6.I strains confirmed the suggestive locus on chr. 12. Some of the QTLs for alcohol consumption are new, while others confirm previously reported QTLs for alcohol preference, and alcohol acceptance.

Discomfort: not pain but still unpleasant feelings from the gut.

Most of the factors initiating food or fluid intake have already been studied, but much less is known about those terminating ingestion. We have hypothesised that discomfort originating from the gastrointestinal system may be one of those factors. Gut distension cause pain if the intestinal volume changes but merely discomfort if only the tension of the gut wall increases. It seems that mild unpleasantness (i.e. discomfort) arising from the gut as a result of moderate (quasi-isometric) distension, among and in concordance with other factors, may significantly reduce intake and hence contribute to physiological satiety. The arising discomfort can be detected by measuring the amount and rate of the ingestion, by recording and analysing ingestive behavior by taste-aversivity and taste-reactivity tests, etc. Conclusions of all experiments point to the same direction: tension increase in the gut wall causes discomfort and results in decrease of intake, i.e. satiety.