Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Schematic representation of the Gs-cAMP pathway. Semaglutide binds the GLP1R and triggers the stimulatory Gs protein to activate adenylyl cyclase (AC). Activated AC then converts adenosine triphosphate (ATP) to cAMP. (b) Virus and injection strategy for expressing Cre recombinase or mCherry control in the dorsal vagal complex of Gnas fl/fl mice. (c) Representative images of viral expression in the three groups of mice: (top) Control (n=9 mice), (middle) DVC ΔGNAS (n=9 mice), and (bottom) AP Miss (n=5 mice). Scale bar = 200 µm. (d) Quantification of extent (%) of Cre viral expression in area postrema (AP) and nucleus of the solitary tract (NTS) in DVC ΔGNAS (square) and AP Miss (triangle) groups. (e) Bodyweight in grams (g) of the three groups of mice over three weeks on high fat diet (HFD) (f) Left: mean change in bodyweight (grams) from baseline on two week daily subcutaneous (SQ) injections of semaglutide (12 µg/kg). Right: bodyweight (%) changes on day 14 of semaglutide treatment. Points represent individual mice. Ordinary One-way ANOVA with Tukey’s multiple comparisons test (**p=0.0045, ****p<0.0001). (g) Simple linear regression of extent of viral expression in either AP (red) or NTS (gray) with the day 14 percent change in bodyweight to semaglutide treatment. AP: slope is significantly different from zero (**p=0.0024). NTS: slope is not significantly different from zero (p=0.6788). Slopes for AP and NTS are significantly different, p=0.0327. (h) Multiple linear regression for weight loss on semaglutide with predictors: β 1 (% of AP viral expression) and β 2 (% of NTS viral expression) (**p=0.0032, ns p=0.5353) (i) Quantification of semaglutide induced Fos in the DVC across AP, NTS, and the dorsal motor nucleus (DMX). Two-way repeated measures ANOVA test with Tukey’s multiple comparisons test (AP:**p=0.0038, *p=0.0241; NTS: *p=0.0443). (j-l) Representative images of Fos antibody staining in DVC quantified in (i) with the control (j) , DVC ΔGNAS (k) , and AP Miss (l) groups. Top: images with 3 colors merged with DAPI in blue, middle: viral expression (magenta), bottom: Fos (green) antibody labeling to semaglutide injection. Scale bar = 200 µm. All data are represented as mean ± S.E.M. See also Extended Data Fig. 1.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Virus, Injection, Expressing, Control, Staining, Antibody Labeling

Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Schematic diagram of transgenic crosses used. Top: IRES-Cre fusion protein is driven under the direction of the Glp1r promoter, middle: CAG promoter and loxP -flanked STOP cassette upstream of the gene encoding soma-targeted GCaMP8s all inserted into the Rosa 26 (R26) locus, bottom: exon 1 of the Gnas gene is flanked by loxP sites. GnasWT mice are a cross of the first two transgenic lines and GnasKO mice are a cross of all three lines. (b) Schematic representation of the 2-photon slice imaging experimental paradigm of calcium transients in Glp1r-expressing DVC slices (Glp1r:soma-GC8s) with Gs intact (GnasWT) or absent (GnasKO). (c) Representative imaging field of view of the AP and NTS in a GnasWT slice. (d) Heatmap of calcium fluorescence of all 2,226 DVC Glp1r cells (GnasWT: n=1,129 cells, 5 slices, 3 mice; GnasKO: n=1,007 cells, 3 slices, 3 mice) ordered by cluster (1-5) and genotype. (e) Average calcium response profile of cells by cluster. (g) Area under the curve quantification during the 10-minuted semaglutide (100 nM) wash per cluster per slice. Repeated measures one-way ANOVA test with Tukey’s multiple comparisons test (*p<0.05, **p<0.01, ***p<0.001, ***p<0.0001). (g) Average response profile by cluster of 2,292 DVC Glp 1r cells (GnasWT: n=1,154 cells, 6 slices, 3 mice; GnasKO: n=1,138 cells, 4 slices, 2 mice) based on their responses to semaglutide. (h) Percentage of cells in each cluster per slice by genotype. Two-way ANOVA with Benjamini, Krieger, and Yekuteli two-stage step-up method to correct for multiple comparisons (cluster 1: *p=0.0230, cluster 3: *p=0.0128). (i-m) Top: average response profile of each clusters response to tetrodotoxin (TTX; 500 nM) application separated by genotype. Bottom: quantification of change in fluorescence in response to TTX application per slice. Two-tailed unpaired t-test (cluster 1: **p=0.0034). (n) Mean calcium response in of semaglutide-responsive GnasWT (black) or GnasKO (red) cells following a 30-minute preincubation with TTX. (o) Area under the curve quantification per slice of bulk calcium fluorescence changes in the AP during the 5-minute baseline period (Bsl), the 10-minute semaglutide period (Sema) and the 5-minute KCl period (KCl) in GnasWT (black; n=4 slices, 2 mice) and GnasKO (red; 2 slices, 2 mice). Two-way ANOVA with Bonferroni’s correction (Sema: *p=0.0247). (p) Mean calcium response in of semaglutide-responsive GnasWT (black) or GnasKO (red) cells in ACSF (solid lines; GnasWT: n=5 slices, 3 mice; GnasKO: n=3 slices, 3 mice) or following a 45-minute preincubation with the Gq inhibitor FR (1 µM) (dotted lines; GnasWT: n=5 slices, 3 mice; GnasKO: n=4 slices, 2 mice). (q) Area under the curve quantification per slice of bulk calcium fluorescence changes in the AP from slices in ACSF (filled bars and points) or in the presence of FR (open bars and points). Bsl: 5-minute baseline period, Sema: 10-minute semaglutide period, KCl: 5-minute KCl period. Two-way ANOVA with Tukey’s multiple comparisons test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). All data are represented as mean ± S.E.M. See also Extended Data Fig. 2 and 3.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Transgenic Assay, Imaging, Expressing, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Virus and injection strategy for expressing cADDis in Glp1r-expressing cells in the AP. (b) Representative image of cADDis expression (green) in the AP. Scale bar = 200 µm. (c) Heatmap of all cells (n= 139 cells, 4 slices, 2 mice) change in fluorescence in response to semaglutide (100 nM) bath application. The y-axis is inverted for clarity, as increases in cAMP are reflected as decreases in fluorescence in the GreenDownwards cADDis sensor. (d) Individual example traces from six cells in response to semaglutide wash (blue square). (e) Average response profile of transient cAMP responders (purple) and sustained cAMP responders (blue). (f) Cumulative frequency distribution of all the fraction of each cells peak response across 5-minute increments. (g) Pie chart of percentage of transient (purple) and sustained (blue) from total cells. (h) Schematic representation of the competitive antagonist exendin 9 (Ex-9) and the agonist semaglutide (Sema) for the GLP1R. (i) Left: average cADDis response profile of all cells (n=384 cells, 6 slices, 4 mice) response to semaglutide (100 nM) bath application followed by Ex-9 (10 µM) bath application in artificial cerebral spinal fluid (ACSF). Right: Pie chart of percentage of sustained (purple) and Ex-9 responsive (blue) and transient (teal) cAMP responders from total cells. (j) Schematic representation of the enzymatic breakdown of cAMP to AMP by phosphodiesterase 4 (PDE4). PDE4 activity is inhibited by Roflumilast. (k) Average cADDis response profile of all cells in response to 15-min bath application of 10 nM semaglutide followed by 15-min bath application of 100 nM semaglutide in ACSF (black line; n=45 cells, 3 slices, 3 mice) or in the presence of Roflumilast (10 µM; red line; n=107 cells, 5 slices, 4 mice). (l) Area under the curve (AUC) quantification per slice. Bsl: 5-minute baseline period, 10 nM: 15-minute 10 nM semaglutide period, 100nM: 15-min 100 nM semaglutide period. Two-way ANOVA with Bonferroni’s multiple comparisons test (**p=0.004). (m) Left: average cADDis response profile of all cells (n=143 cells, 3 slices, 3 mice) response to semaglutide (100 nM) bath application followed by Ex-9 (10 µM) bath application in the presence of Roflumilast (10 µM). Right: pie chart of percentage of sustained (purple) and Ex-9 responsive (blue) cAMP responders from total cells. (n) Percentage of cAMP type responses (sustained, ex-9 responsive or transient) per slice in ACSF (black) or Roflumilast (red). Two-way ANOVA with Bonferroni’s multiple comparisons test (Sustained: *p=0.0119, Ex-9 responsive: *p=0.0386, Transient: *p=0.0487). (o) Virus and injection strategy for expressing cADDis and Cre-recombinase in the AP of Gnas fl/fl mice to measure cAMP responses in the absence of Gs. (p) Left: average response profiles of semaglutide-responders (Sema) or forskolin only (Fsk only) responders (n= 422 cells, 7 slices, 3 mice). Right: pie chart of percentage of Fsk only and Sema cAMP responders from total cells. All data are represented as mean ± S.E.M. See also Extended Data Fig. 4.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Virus, Injection, Expressing, Fluorescence, Activity Assay

Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Virus and injection strategy for expressing PDE4-cat with cADDis or GCaMP7s in Glp1r-expressing cells in the AP. (b) Schematic representation of the Gs-cAMP pathway. Semaglutide binds the GLP1R and triggers the stimulatory Gs protein to activate adenylyl cyclase (AC). Activated AC then converts adenosine triphosphate (ATP) to cAMP, which leads to downstream neuronal activation. cAMP is degraded by the constitutively active PDE4-cat. (c) Representative images of viral expression of PDE4-cat (magenta; right) and cADDis (green; middle) in AP Glp1r cells. Scale bar = 200 µm. (d) Average response profiles of all cells cADDIs response to 10-minute bath application of semaglutide (1 µM) in cells lacking PDE4-cat expression (Transient: n=56 cells, 3 slices, 3 mice or Sustained: n=81 cells, 3 slices, 3 mice) or with PDE4-cat expression (n=140 cells, 3 slices, 3 mice). (e) Area under the curve (AUC) quantification per slice during 5-minute Baseline period and 10-minute Semaglutide (Sema) period. Two-way ANOVA with Tukey’s multiple comparisons test (****p<0.0001). (f) Representative image of viral expression of PDE4-cat (magenta) and GCaMP7s (green) in AP Glp1r cells. Scale bar = 200 µm. Yellow box indicates field of view in (G). (g) Representative image of viral expression from yellow box in (F) showing PDE4-cat (magenta; right) colocalization with GCaMP7s (green; middle). White arrows indicate colocalization (PDE4+) and yellow arrows indicate GCaMP7s lacking colocalization (PDE4-). Scale bar = 100 µm. (h) Average response profiles of all cells GCaMP7s response to 10-minute bath application of semaglutide (1 µM) and 5-minute application of 10 mM KCl in cells lacking PDE4-cat expression (PDE4-: n=59 cells, 4 slices, 4 mice) or with PDE4-cat expression (PDE4+: n=102 cells, 4 slices, 4 mice). (i) Area under the curve (AUC) quantification per slice during 5-minute Baseline period , 10-minute Semaglutide (Sema) period and 5-minute 10 mM KCl period. Two-way ANOVA with Bonferroni’s multiple comparisons test (***p= 0.0009). (j) Heatmap of all GCaMP7s-expressing cells fluorescence response ordered by PDE4- (black) and PDE4+ (magenta). Dotted white lines indicate times of solution change. All data are represented as mean ± S.E.M.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Virus, Injection, Expressing, Activation Assay, Fluorescence

Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Virus and injection strategy for expressing PDE4-cat in the DVC of Glp1r-ires-cre mice. (b) Representative histological image of virus expression in Glp1r-expressing cells in the DVC with DAPI (blue) and PDE4-cat (red). Scale bar = 200 µm. (c) Changes in bodyweight in grams (left), week 5 percentage bodyweight (middle), and food intake (right) in the control (black; n=15 mice, 8 males) and PDE4 (blue; n=10 mice, 8 males) group on 5 weeks of high fat diet. Multiple unpaired t-tests with Welch’s correction (left: ****p<0.0001, right: *p<0.0001). (d) Left: change in bodyweight in the control group and PDE4 group in response to two weeks of daily SQ injection of semaglutide (12 µg/kg) or saline. Right: Day 14 change in bodyweight as percent. One-way ANOVA with Tukey’s multiple comparison’s test (****p<0.0001). (e) Left: Daily food intake in grams of the control group and PDE4 group in response to two weeks of daily SQ injection of semaglutide (12 µg/kg) or saline. Right: Mean food intake per day across 14 days of treatment. Brown-Forsythe and Welch’s one-way ANOVA with Dunnett’s T3 multiple comparisons test (*p= 0.0204, Control Sema vs PDE4 Saline:**p= 0.0097, PDE4 Sema vs Control Saline:**p= 0.0085, ***p= 0.0004, ****p<0.0001). (f-g) (f) Axonal density quantification of projections from AP Glp1r neurons expressing mKate2-PDE4-cat in representative images found in (g) . Points represent individual mice. (g) elPBN: external lateral parabrachial nucleus; aBNST: anterior bed nucleus of the stria terminals; PVH: paraventricular hypothalamus; PVT: paraventricular thalamus; ARC: arcuate hypothalamus; scp: superior cerebellar peduncle; aco: anterior commissure; 3V: third ventricle. All scale bars = 200 µm. Repeated measures ANOVA with Tukey’s multiple comparisons test (elPBN vs. all other brain regions****p<0.0001). (h-i) Representative images of Fos immunostaining in response to semaglutide injections in mice expressing PDE4-cat in DVC Glp1r neurons (h) or in Control mice (i) . (H) Left: PDE4-cat (magenta), semaglutide induced Fos (green), and DAPI (blue) merged. Right: Fos and DAPI of the same representative images from the right. All scale bars = 200 µm. (j) Quantification of brain-wide Fos expression in response to semaglutide in control (black) and PDE4 (blue) mice. Multiple unpaired t-test with Welch’s correction using the Holm-Šídák method (AP: p<0.000001, NTS: *p= 0.0379, elPBN: *p= 0.00016, CeA: *p=0.0379). (k) Schematic depiction that semaglutide binds and activates GLP1Rs in the AP. AP Glp1r cells send downstream projections to recruit elPBN and indirectly recruit activation of the CeA. All data are represented as mean ± S.E.M.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Virus, Injection, Expressing, Control, Saline, Immunostaining, Activation Assay

Journal: bioRxiv

Article Title: Semaglutide drives weight loss through cAMP-dependent mechanisms in GLP1R-expressing hindbrain neurons

doi: 10.1101/2025.08.12.668772

Figure Lengend Snippet: (a) Virus and fiber implant strategy for expressing GCaMP7s in the elPBN of FosTRAP2.0-cre mice to measure the activity of semaglutide-activated elPBN neurons. (b) Representative histological image of virus expression and fiber implant location above the elPBN with DAPI (blue) and GCaMP7s (green) from an elPBN SemaTRAP mouse. Scale bar=200 µm. (c) Mean fiber photometry bulk calcium fluorescence (z-score) from elPBN semaTRAP neurons (n=5 mice) in response to SQ saline (gray) or SQ semaglutide (blue; 120 µg/kg) injection. (d) Area under the curve quantification from time 0 to time 20 post injection. Gray lines represent individual mice. Two-tailed paired t-test (***p=0.001). (e) Virus and injection strategy for expressing TeNT bilaterally in the elPBN of FosTRAP2.0-cre mice. (f) Representative histological images of DAPI (blue) and TeNT viral expression (green) in the elPBN in VehTRAP (top) or SemaTRAP (bottom) mice. All scale bars = 200 µm. (h-g) Conditioned taste aversion behavioral paradigm with daily fluid intake (h) and ensure preference ratio (g) of five groups of mice. Control: Vehicle paired novel flavor group (n=6 mice); Semaglutide: Semaglutide (12 µg/kg) paired novel flavor group (n=5 mice); VehTRAP: Semaglutide (12 µg/kg) paired novel flavor with TeNT expressed in elPBN VehTRAP neurons (n=6 mice); SemaTRAP: Semaglutide (12 µg/kg) paired novel flavor with TeNT expressed in elPBN SemaTRAP neurons (n=8 mice); Sema/sema: Semaglutide (12 µg/kg) paired novel flavor in mice with previous exposure to semaglutide (n=5 mice). One-way ANOVA with Tukey’s multiple comparisons test (**p= 0.0028, ***p= 0.0001; ****p<0.0001). (i) Bodyweight (grams) of mice on Day 0 of semaglutide treatment in (j). One-way ANOVA with Tukey’s multiple comparisons test (ns: not significant). (j) Left: mean weight responses to two weeks of daily SQ semaglutide (12 µg/kg) treatment in Control mice (n=11 mice), elPBN VehTRAP mice (n=6 mice) and elPBN SemaTRAP mice (n=6 mice). Right: change in bodyweight on Day 14 of semaglutide treatment. Points represent individual mice. One-way ANOVA with Tukey’s multiple comparisons test (*p=0.0191, **p=0.002). All data are represented as mean ± S.E.M. See also Extended Data Fig. 5.

Article Snippet: The following drugs were used for this study: Semaglutide (Adipogen Life Siences; Cat # CAS0910463-68-2), Forskolin (Tocris; Cat # 1099), Tetrodotoxin citrate (Tocris; Cat # 1069), Roflumilast (Sigma Aldrich; Cat # 1099), and FR900359 (Cayman Chemical Company; Cat # 33666)

Techniques: Virus, Expressing, Activity Assay, Fluorescence, Saline, Injection, Two Tailed Test, Control

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. High (1.0 mg/kg), medium (0.3 mg/kg) and low (0.1 mg/kg) doses of LY37 treatment have no effect on escape latencies in SAL rats (n = 6–12 for each group). B. only 0.3 mg/kg LY37 treatment improves learning ability in MAM rats. There are significant shorter latencies on day 4 and day 5 in MAM-LY37 (0.3 mg/kg) rats compared with MAM-SAL rats (n= 6–16 for each group). C. MAM rats exhibited significantly longer escape latencies from training day 3 to day 5 in water maze (replotted from A and B). D. In probe test, MAM rats show decreased time in the target quadrant compared with SAL rats. There is a significant increase in time spent on target quadrant in MAM-LY37 (0.3 mg/kg) rats compared with their saline controls. A–C, three-way repeated measure ANOVA followed by Bonferroni's test. D, two-way ANOVA followed by Bonferroni's test.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques:

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. Representative western blots and summary histograms show the NMDAR subunit and mGluR2/3 protein levels in the mPFC from SAL-SAL, SAL-LY37 (0.3 mg/kg), MAM-SAL, and MAM-LY37 rats 24 hours after probe test of water maze (n = 6). MAM exposure resulted in significant decreases of GluN2B and mGluR2, and the levels of GluN2B, but not mGluR2, are rescued by LY37 treatment. B. Representative western blots and summary histograms show the levels of Akt-GSK3β-mTOR signaling after probe test. No significant difference was detected for the Akt1, pAktSer473, mTOR or pmTORSer2448 among groups. MAM exposure resulted in reduced pGSK3βSer9 levels, which are restored by LY37 treatment. * p < 0.05, ** p < 0.01 two-way ANOVA followed by Bonferroni's test.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Western Blot

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. LY37 (300 nM, bath application) significantly increased PPR and decreased CV−1 of evoked NMDA-EPSCs in SAL group (n = 9 from 3 rats), indicating a decrease of glutamate release. B. In contrast, LY37 had no effect on both PPR and CV−1 in MAM rats (n = 9 from 3 rats), suggesting a blunted presynaptic response to mGlu2/3 agonists. * p < 0.05, ** p < 0.01 two-way ANOVA followed by Bonferroni's test

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques:

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A and B. Representative eEPSC traces (A) and summarized input-output curves (B) of NMDAR-mediated EPSC in response to a series of stimulation intensities in SAL-SAL, SAL-LY37, MAM-SAL and MAM-LY37 rats (n = 9–10 cells from 3 rats for each group). In all three intensities, MAM rats show decreased NMADR-eEPSCs compared with SAL-SAL and MAM-LY37 groups. * p < 0.05 vs. SAL-SAL, # p < 0.01 vs. MAM-LY37, two-way RM ANOVA followed by Bonferroni's test. C. Representative NMDAR-eEPSC traces at baseline and after Ro25-6981 (Ro, 1 µM) bath application. The lower panel of scaled traces shows the reduced decays caused by Ro application as indicated by shadow area. D. Summary histograms show that there is significant decrease of NMDAR-eEPSC amplitude in SAL-SAL (n = 12 cells from 4 rats), SAL-LY37 (n = 10 cells from 3 rats) and MAM-LY37 (n = 11 cells from 4 rats) groups after Ro application, but not that in MAM-SAL rats (n = 11 cells from 4 rats for each group). E. Summary graph shows a significant decrease of Ro-sensitive currents of NMDA channels in MAM-SAL but not in MAM-LY37 rats. * p < 0.05, ANOVA followed by Bonferroni's test. Data are mean ± SEM.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques:

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A and B. Representative NMDAR-EPSC original (upper) and scaled (lower) traces in SAL (A) and MAM group (B). Summary histograms show the decreases of amplitudes caused by LY37 are less affected by loading pGSK3βSer9 antibody (pGSK3β Ab, 10 mg/ml) into the recording pipettes in SAL group (n = 13 from 5 rats without pGSK3βAb, n = 12 from 5 rats with pGSK3βAb), but the increased decay time is blocked by postsynaptic pGSK3βSer9 inhibition. For MAM rats, No significant difference of amplitudes before and after LY37 with or without pGSK3βSer9 Ab (n = 13 from 5 rats without pGSK3βAb, n = 13 from 5 rats with pGSK3βAb), whereas increased decay time is blocked by postsynaptic pGSK3βSer9 inhibition. * p < 0.05, *** p < 0.001, two-way RM ANOVA followed by Bonferroni's test. Data are mean ± SEM.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Inhibition

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. During the LY37 treatment (P21 to P27), no difference in the body weight among SAL-SAL, SAL-LY37, MAM-SAL and MAM-LY37 (n = 9 for each group) is found. Represent images (B) of Golgi staining of mPFC showing that the decreased apical dendritic spine density in layer 5 pyramidal neurons from MAM rats is rescued by early LY37 treatment (C). Scale bars = 50 µm for the upper panel and 10 µm for the lower panel. D. No difference was found in mushroom, stubby, and thin spines fraction among SAL-SAL, MAM-SAL, SAL-LY37 and MAM-LY37 groups (n = 8 neurons from 4 rats). ***p < 0.001, ANOVA followed by Bonferroni's test. Data are mean ± SEM.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Staining

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. Representative traces of 10 stimuli evoked EPSPs from SAL-SAL (n=10 cells from 3 rats), SAL-LY37 (n=9 cells from 3 rats), MAM-SAL (n=9 cells from 3 rats), and MAM-LY37 (n=9 cells from 3 rats) groups. The scaled traces show the area under the EPSPs, the differences between saline-treatment and LY37 treatment are highlighted by the shadow area. B. Summary line graphs show that there is a decrease of eEPSP amplitude in MAM-SAL rats that can be correct by juvenile LY37 treatment. * p < 0.05, *** p < 0.001 vs. SAL-SAL, # p < 0.05 vs. MAM-LY37, two-way RM ANOVA followed by Bonferroni's test. Data are mean ± SEM. C. The area under EPSPs of MAM-SAL rats is significantly smaller than SAL-SAL group and MAM-LY37 group. * p < 0.05, ** p < 0.01, Kruskal-Wallis test followed by Dunn's test, Data are median ± interquartile range.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques:

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: A. MAM rats showed impaired spatial memory assessed by spontaneous T-maze. These deficits were absent in the MAM rats with juvenile LY37 treatment. ** p < 0.01, # p < 0.05, Kruskal-Wallis test followed by Dunn's test, Data are median ± interquartile range. B. MAM animals exhibited a significantly increased spontaneous locomotion activity (arrowhead) in response to MK801 administration (0.05 mg/kg). Juvenile LY37 treatment normalized the locomotor activity in response to either novel environment (first 30 min) or MK801 (rest 60 min). * p < 0.05 MAM-SAL vs. SAL-SAL, # p < 0.05 MAM-LY37 vs. SAL-SAL, two-way RM ANOVA followed by Bonferroni's test. Data are mean ± SEM.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Activity Assay

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: Summary graphs of action potentials from saline- and LY37-treated rats 1 hour (A), 1 week (B), 5 weeks (C) and 10 weeks (D) (n = 13 to 20 from 6–7 rats) after the last injection. Summary histograms of amplitude (E) and frequency (F) of spontaneous EPSCs recorded at −70 mV in layer 5 pyramidal neurons from normal juvenile rats treated with LY37 (0.3 mg/kg/day, i.p., once daily, P21–27) 1 hour, 1 week, 5 weeks and 10 weeks after the last injection. A–D, * p < 0.05, two-way RM ANOVA followed by Bonferroni's test. Data are mean ± SEM. E–F, * p < 0.05, unpaired student t-test.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Injection

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: Physiological Properties of layer V Pyramidal Neurons in normal rats after repeated SAL- or LY37 exposure

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques:

Journal: Neuropharmacology

Article Title: Juvenile treatment with mGluR2/3 agonist prevents schizophrenia-like phenotypes in adult by acting through GSK3β

doi: 10.1016/j.neuropharm.2018.05.019

Figure Lengend Snippet: In normal condition, glutamate release from the presynaptic site, and activation of presynaptic mGluR2/3 inhibits glutamate release. Postsynaptic mGluR2/3, however, activates GSK3β by decreasing the inhibitory pGSK3βSer9, leading to an increase of an NMDA-GluN2B function. The overall effect of LY37 depends on the dosage. In contrast, the reduction of presynaptic mGluR2 in MAM animals reduces the effect of LY37 on inhibition of glutamate release, whereas the postsynaptic enhancement of NMDAR-GluN2B modulated by postsynaptic mGluR2/3 retains. Treatment with LY37 during juvenile period rescues the GluN2B-GSK3β signaling, improves the NMDAR hypofunction, and thus prevents the consequential morphological changes and behavioral deficits.

Article Snippet: {"type":"entrez-nucleotide","attrs":{"text":"LY379268","term_id":"1257807854","term_text":"LY379268"}} LY379268 (LY37, Cat. No. 2453), Ro 25-6981 (Cat. No. 1594/1), and tetrodotoxin (TTX) (Cat. No. 1078/1) were obtained from Tocris Bioscience.

Techniques: Activation Assay, Inhibition