Journal: Journal of Neuroscience

Article Title: Inactivation of Socs3 in the Hypothalamus Enhances the Hindbrain Response to Endogenous Satiety Signals via Oxytocin Signaling

doi: 10.1523/jneurosci.1669-12.2012

Figure Lengend Snippet: Figure 1. Efficiency of Cre-mediated recombination and validation of functional Socs3 inactivation. A, Representative pictures showing typical -gal immunohistochemical staining (red) obtained 2 weeks after Ad-Cre delivery in the MBH of 3-month-old, ROSA26R male mice. Cre-mediated recombination is extensively detected in Hu C/D-positive neurons (green) in the ARC, VMH, andDMH(top).However,virtuallyno-galstainingisdetectedinthePVN(bottom).3V,3rdventricle;AHP,anteriorhypothalamicarea,posteriorpart;DM,dorsomedialhypothalamicnucleus;ME, medianeminence;VMHDM,dorsomedialpartoftheVMH;VMHVL,ventrolateralpartoftheVMH.B,NeuronstransducedwithAd-CreintheMBHshowleptin-inducedStat3Y705-phosphorylation (pStat3,brown)inmiceinjectedwithleptin(1mg/kg;i.p.)andperfused1hlater.TransducedneuronswerealsostainedwithX-gal(blue)toreveal-galactivity.C,D,WesternblotanalysesofSocs3 and-actininMBHlysatesofWTandSocs3 MBHmice.Micewerefastedovernightandtissueswereprocessed5haftervehicle(Veh)orleptininjection(3.5mg/kg;i.p.).EachlaneinCrepresents one animal. In D, Socs3 expression normalized to -actin levels, quantified by densitometry (n 6–8 animal per group). Data are mean SEM **p 0.004, Mann–Whitney test. NS, nonsignificant. Scale bars: 100 m.

Article Snippet: Membranes were then blocked in 20 mM Tris buffer, pH 7.4, containing 5% bovine serum albumin and 0.1% Tween 20 and incubated with a rabbit polyclonal anti-Socs3 antibody (1:1000; Cell Signaling Technology) overnight at 4°C.

Techniques: Biomarker Discovery, Functional Assay, Immunohistochemical staining, Staining, Phospho-proteomics, Expressing, MANN-WHITNEY

Journal: Journal of Neuroscience

Article Title: Inactivation of Socs3 in the Hypothalamus Enhances the Hindbrain Response to Endogenous Satiety Signals via Oxytocin Signaling

doi: 10.1523/jneurosci.1669-12.2012

Figure Lengend Snippet: Figure 3. MBH Socs3-deficient mice display an increased response to exogenous leptin. Leptin sensitivity was determined by measuring 24 h food intake on the second day of treat- ment with exogenous leptin (1 mg/kg; i.p.), administered twice daily (n 8 WT and n 10 Socs3 MBH mice). Data are mean SEM. *p 0.05 and ***p 0.001, determined by Bonferronimultiple-comparisontestfollowingrepeated-measures,two-wayANOVA.NS,non- significant; Veh, vehicle; Lep, leptin.

Article Snippet: Membranes were then blocked in 20 mM Tris buffer, pH 7.4, containing 5% bovine serum albumin and 0.1% Tween 20 and incubated with a rabbit polyclonal anti-Socs3 antibody (1:1000; Cell Signaling Technology) overnight at 4°C.

Techniques:

Journal: Journal of Neuroscience

Article Title: Inactivation of Socs3 in the Hypothalamus Enhances the Hindbrain Response to Endogenous Satiety Signals via Oxytocin Signaling

doi: 10.1523/jneurosci.1669-12.2012

Figure Lengend Snippet: Figure 4. MBH Socs3 deficiency triggers an increased hindbrain response to endogenous meal-relatedsatietysignals.A,Cumulativeshort-termfoodintakeatthebeginningofthedark phase measured in nonfasted WT (n 11) and Socs3 MBH mice (n 10) at 30 min, 1 h, and 2hafterlightsoff.B,Nocturnal,2hfoodintakeinnonfastedWT(n12)andSocs3 MBHmice (n9)measuredafterinjectionofvehicle(Veh)ordevazepide(1mg/kg;i.p.),anantagonistof CCK-Areceptors.C,Afteranovernightfast,30minfoodintakewasmeasuredinWT(n8)and Socs3 MBH mice (n 6) injected with leptin (Lep; 0.5 mg/kg; i.p.) or vehicle (Veh), in combi- nationwithCCK(2g/kg;i.p.)orvehicle.DataaremeanSEM.Two-way,repeated-measures ANOVA, followed by Bonferroni multiple-comparison test was used in A–C, where *p 0.05 and **p 0.01. In C, ##p 0.01 versus vehicle-treated Socs3 MBH mice, determined by Bonferroni multiple-comparison test following two-way, repeated-measures ANOVA. NS, nonsignificant.

Article Snippet: Membranes were then blocked in 20 mM Tris buffer, pH 7.4, containing 5% bovine serum albumin and 0.1% Tween 20 and incubated with a rabbit polyclonal anti-Socs3 antibody (1:1000; Cell Signaling Technology) overnight at 4°C.

Techniques: Injection, Comparison

Journal: Journal of Neuroscience

Article Title: Inactivation of Socs3 in the Hypothalamus Enhances the Hindbrain Response to Endogenous Satiety Signals via Oxytocin Signaling

doi: 10.1523/jneurosci.1669-12.2012

Figure Lengend Snippet: Figure 5. Hindbrain oxytocin signaling mediates the increased sensitivity to satiety signals in Socs3 MBH mice. A, Nocturnal food intake in nonfasted WT (n 6) and Socs3 MBH mice (n 7) measured 2 h after 4V injection of vehicle (Veh) or SSR126768A (SSR), an oxytocin receptor antagonist. B, Mature oxytocin tissue content in the DVC of WT and Socs3 MBH mice (n 8 or n 7 in each genotype for the vehicle or leptin-treated groups, respectively) sub- jected to an overnight fast was determined by enzyme immunoassay 5 h after injection of vehicle (Veh) or leptin (3.5 mg/kg; i.p.). Data are mean SEM. *p 0.05 and **p 0.01, determined by Bonferroni multiple-comparison test following two-way, repeated-measures ANOVAinAortwo-wayANOVAinB.InA, ##p0.01versusvehicle-treatedSocs3 MBHmice, determined by Bonferroni multiple-comparison test following two-way, repeated-measures ANOVA.

Article Snippet: Membranes were then blocked in 20 mM Tris buffer, pH 7.4, containing 5% bovine serum albumin and 0.1% Tween 20 and incubated with a rabbit polyclonal anti-Socs3 antibody (1:1000; Cell Signaling Technology) overnight at 4°C.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Neuroscience

Article Title: Inactivation of Socs3 in the Hypothalamus Enhances the Hindbrain Response to Endogenous Satiety Signals via Oxytocin Signaling

doi: 10.1523/jneurosci.1669-12.2012

Figure Lengend Snippet: Figure6. Oxytocinrelayshypothalamicactionofleptinontohindbrainsatietycircuits.Schematicrepresentationoftherodent brainshowingtwomajorbrainregionsinvolvedinthecontrolofenergyhomeostasis:thehypothalamus(yellow)andtheNTS(red) in the caudal brainstem. The hypothalamus receives adipose signals such as leptin, and is involved in the long-term regulation of energy balance by fine-tuning various biological processes, including meal size, which is controlled by satiety circuits in the NTS. Here,inactivationofSocs3selectivelyintheMBHofadultmicerevealsacriticalfunctionofSocs3intheregulationofenergybalance in physiological conditions, as this reduced food intake and protected against body weight gain. This Socs3-dependent reduction offoodintakeisexplainedbyanincreasedsensitivitytoendogenous,meal-relatedsatietysignals,mediatedbyoxytocinsignaling inthecaudalbrainstem.WeconcludethatoxytocinlikelyrelayshypothalamicactionofleptinontoNTSsatietycircuits.Thesedata provideananatomicalsubstratefortheeffectofleptinonmealsize,andmoregenerally,amechanismforhowthebraincontrols meal size as a function of the energetic stores available in the organism to maintain energy homeostasis.

Article Snippet: Membranes were then blocked in 20 mM Tris buffer, pH 7.4, containing 5% bovine serum albumin and 0.1% Tween 20 and incubated with a rabbit polyclonal anti-Socs3 antibody (1:1000; Cell Signaling Technology) overnight at 4°C.

Techniques: