Journal: Journal of Neuroscience

Article Title: Binge Drinking Upregulates Accumbens mGluR5-Homer2-PI3K Signaling: Functional Implications for Alcoholism

doi: 10.1523/jneurosci.5900-08.2009

Figure Lengend Snippet: Figure 3. Repeated bouts of binge alcohol intake with the SHAC procedure elevate NAC protein expression of Homer2, NR2 subunits, and PI3K activation in B6 mice. a, Representative immunoblots for the total protein levels of Homer2a/b, Homer1b/c, mGluR1, mGluR5, NR2a, NR2b, PI3K, p(Tyr)p85 PI3K binding motif [p(Try)p85], and calnexin (loading control) in the NAC of groupsofmicekilledat24haftersixboutsof30minaccessto5%alcohol(SHAC)orwater.b,Summaryofthechangeinprotein expressionafterwithdrawalfromsixboutsoflimitedaccesstoalcohol,expressedasapercentageoftheaverageproteinexpres- sion of water-drinking controls. Compared with water controls, binge alcohol intake with the SHAC procedure significantly increased Homer2a/b, NR2a/b, and p(Tyr)p85 expression (t tests, *p 0.05). Data in b represent the mean SEM of the number of animals indicated in the figure.

Article Snippet: The following rabbit polyclonal antibodies were used: anti-Homer 2a/b and anti-Homer 1b/c (1:1000 dilution; Dr. Paul F. Worley, Johns Hopkins University School of Medicine, Baltimore, MD), anti-mGluR5 (1:1000 dilution; Upstate Cell Signaling Solutions), antiNR2a and anti-NR2b (1:1000 dilution; Calbiochem), anti-PI3K antibody (1:1000 dilution; Upstate Cell Signaling Solutions), and anti-p(Tyr)p85 PI3K binding motif (1:500; Cell Signaling Technology).

Techniques: Expressing, Activation Assay, Western Blot, Binding Assay, Control

Journal: Journal of Neuroscience

Article Title: Binge Drinking Upregulates Accumbens mGluR5-Homer2-PI3K Signaling: Functional Implications for Alcoholism

doi: 10.1523/jneurosci.5900-08.2009

Figure Lengend Snippet: Figure 5. Blockade of NAC mGluR5 and PI3K, but not mGluR1, reduces binge alcohol drinking in B6 mice. a, Summary of the effectsofanintra-NACshellinfusionofvariousdosesofthemGluR5antagonistMPEP(filledbars),50ng/sidewortmannin,andthe combinationofMPEPpluswortmannin(hatchedbars),aswellasvariousdosesofthemGluR1antagonistCPCCOEt(openbars)on 5%alcoholintakeduringa30minperiodwiththeSHACprocedure.b,Summaryoftheeffectsofintra-NACMPEPandwortmannin onwaterintakeduringa30minperiod.ThedatarepresentthemeanSEMofthenumberofanimalsindicatedineachbarinthe figure. *p 0.05 versus respective vehicle pretreatment (i.e., 0 dose) (least significant difference post hoc tests).

Article Snippet: The following rabbit polyclonal antibodies were used: anti-Homer 2a/b and anti-Homer 1b/c (1:1000 dilution; Dr. Paul F. Worley, Johns Hopkins University School of Medicine, Baltimore, MD), anti-mGluR5 (1:1000 dilution; Upstate Cell Signaling Solutions), antiNR2a and anti-NR2b (1:1000 dilution; Calbiochem), anti-PI3K antibody (1:1000 dilution; Upstate Cell Signaling Solutions), and anti-p(Tyr)p85 PI3K binding motif (1:500; Cell Signaling Technology).

Techniques:

Journal: Journal of Neuroscience

Article Title: Binge Drinking Upregulates Accumbens mGluR5-Homer2-PI3K Signaling: Functional Implications for Alcoholism

doi: 10.1523/jneurosci.5900-08.2009

Figure Lengend Snippet: Figure7. mGluR5 F1128RmutationreducesNACbasalPI3Kactivity.a,Representativeimmunoblotsforthetotalproteinlevels of Homer2a/b, Homer1b/c, mGluR1, mGluR5, NR2a, NR2b, PI3K, p(Tyr)p85 PI3K binding motif [p(Try)p85], and calnexin (loading control) in the NAC of experimentally naive WT and mGluR5 F1128R TG mice. b, Summary of the genotypic differences in proteinexpression,expressedasapercentageofaveragelevelsofWTcontrols.DatainbrepresentthemeanSEMofthenumber of mice indicated in the figure. *p 0.05 versus WT (t tests).

Article Snippet: The following rabbit polyclonal antibodies were used: anti-Homer 2a/b and anti-Homer 1b/c (1:1000 dilution; Dr. Paul F. Worley, Johns Hopkins University School of Medicine, Baltimore, MD), anti-mGluR5 (1:1000 dilution; Upstate Cell Signaling Solutions), antiNR2a and anti-NR2b (1:1000 dilution; Calbiochem), anti-PI3K antibody (1:1000 dilution; Upstate Cell Signaling Solutions), and anti-p(Tyr)p85 PI3K binding motif (1:500; Cell Signaling Technology).

Techniques: Binding Assay, Control

Journal: Journal of Neuroscience

Article Title: Binge Drinking Upregulates Accumbens mGluR5-Homer2-PI3K Signaling: Functional Implications for Alcoholism

doi: 10.1523/jneurosci.5900-08.2009

Figure Lengend Snippet: Figure 8. Line differences in NAC Homer2 and PI3K activity in selectively bred SHAC and SLAC mice. a, b, Summary of the averagealcoholdoseconsumed(gramsperkilogram;a)andBACs(b)attainedafterthesecondsessionof30minalcoholdrinking (selection phenotype) in the genetically heterogeneous HS/Npt mice (foundation population for the selection) and in the SHAC andSHACoffspring,acrossfourgenerationsofselectivebreeding.DatainaandbrepresentthemeanSEMof80–104miceper line per generation; *p 0.05, SHAC offspring versus SLAC offspring. c, Representative immunoblots for the total protein levels of Homer2a/b, Homer1b/c, mGluR1, mGluR5, NR2a, NR2b, PI3K, p(Tyr)p85 PI3K binding motif [p(Try)p85], and calnexin (loadingcontrol)intheNACoffourthgeneration(S4)selectivelybredSHACandSLACmice,at3monthsaftertheirsecond30min alcohol-drinkingsession.d,Summaryofthelinedifferencesinproteinexpression,expressedasapercentageofaveragelevelsof SLACanimals.DataindrepresentthemeanSEMofthenumberofmiceindicatedinthefigure.*p0.05versusSLAC(ttests).

Article Snippet: The following rabbit polyclonal antibodies were used: anti-Homer 2a/b and anti-Homer 1b/c (1:1000 dilution; Dr. Paul F. Worley, Johns Hopkins University School of Medicine, Baltimore, MD), anti-mGluR5 (1:1000 dilution; Upstate Cell Signaling Solutions), antiNR2a and anti-NR2b (1:1000 dilution; Calbiochem), anti-PI3K antibody (1:1000 dilution; Upstate Cell Signaling Solutions), and anti-p(Tyr)p85 PI3K binding motif (1:500; Cell Signaling Technology).

Techniques: Activity Assay, Selection, Western Blot, Binding Assay