Journal: Frontiers in Pharmacology

Article Title: Activation of GLP-1R ameliorates alcohol withdrawal induced anxiety-like behavior by regulating neuronal mitochondrial quality control

doi: 10.3389/fphar.2026.1820128

Figure Lengend Snippet: Effects of CEE on anxiety-like behaviors and GLP-1R expression in mice. (A) Representative tracking plot from the OFT. (B) Number of entries in the center area (**p = 0.0015. n = 9 mice per group). (C) Time spent in the center area (**p = 0.0056. n = 9 mice per group). (D) Representative track plot of the EPM test. (E) Number of entries in the open arms (**p = 0.0070. n = 9 mice per group). (F) Time spent in the open arms during the EPM test (**p = 0.0056. n = 9 mice per group). (G,H) Protein expression of GLP-1R was detected by Western blot analysis (***p = 0.0002. n = 6 mice per group). (I) Representative images of Immunofluorescence staining of GLP-1R and NeuN with DAPI nuclear counterstaining in the PFC. *p = 0.0113. n = 3 mice per group). NeuN (green) was used to label mature neurons, and DAPI (blue) was used as a nuclear counterstain. GLP-1R (red) was predominantly co-localized with NeuN-positive neurons, indicating neuronal expression. Scale bar = 100 μm. (J) PFC dissection: bregma +1.69 mm. All data are presented as mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: CEE, Chronic ethanol exposure; GLP-1R, Glucagon-like Peptide-1 receptor; OFT, Open Field Test; EPM, Elevated Plus Maze.

Article Snippet: Immunofluorescence staining was performed using primary antibodies against GLP-1R ( GB113881 , 1:5000, Servicebio) and NeuN (GB11138, 1:5000, Servicebio).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Dissection

Journal: Frontiers in Pharmacology

Article Title: Activation of GLP-1R ameliorates alcohol withdrawal induced anxiety-like behavior by regulating neuronal mitochondrial quality control

doi: 10.3389/fphar.2026.1820128

Figure Lengend Snippet: Effects of Semaglutide on CEE-induced anxiety-like behaviors and GLP-1R expression. (A) Representative tracking plot from the OFT. (B) Number of entries in the center area (**p < 0.01. n = 19 mice per group). (C) Time spent in the center area (***p < 0.001. n = 19 mice per group). (D) Representative track plot of the EPM test. (E) Number of entries in the open arms (***p < 0.001. n = 19 mice per group). (F) Time spent in the open arms during the EPM test (*p < 0.05, ***p < 0.001. n = 19 mice per group). (G,H) Protein expression of GLP-1R was detected by Western blot analysis (**p < 0.01. n = 6 mice per group). (I) Representative confocal images of Immunofluorescence staining of GLP-1R and NeuN with DAPI nuclear counterstaining in the PFC. (*p < 0.05, **p < 0.01. n = 4 mice per group). NeuN (green) was used to label mature neurons, and DAPI (blue) was used as a nuclear counterstain. GLP-1R (red) was predominantly co-localized with NeuN-positive neurons, indicating neuronal expression. Scale bar = 100 μm. (J) PFC dissection: bregma +1.69 mm. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: CEE, Chronic ethanol exposure; GLP-1R, Glucagon-like Peptide-1 receptor; OFT, Open Field Test; EPM, Elevated Plus Maze; Sema, Semaglutide.

Article Snippet: Immunofluorescence staining was performed using primary antibodies against GLP-1R ( GB113881 , 1:5000, Servicebio) and NeuN (GB11138, 1:5000, Servicebio).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Dissection

Journal: Frontiers in Pharmacology

Article Title: Activation of GLP-1R ameliorates alcohol withdrawal induced anxiety-like behavior by regulating neuronal mitochondrial quality control

doi: 10.3389/fphar.2026.1820128

Figure Lengend Snippet: Effects of Semaglutide on mitochondrial damage in CEE mice. (A,B) Protein expression of mitochondrial GLP-1R was detected (*p < 0.05, ***p < 0.001. n = 6 mice per group). (C,D) Protein expression of mitochondrial CREB was detected by Western blot analysis (*p < 0.05, **p < 0.01. n = 6 mice per group). (E-J) Protein expression of mitochondrial OXPHOS was detected by Western blot analysis, including complex I (NDUFB8), complex II (SDHB), complex III (UQCRC2), complex IV (MTCO1), and complex V (ATP5A) (*p < 0.05, **p < 0.01. n = 6 mice per group). (K) Mitochondrial morphology of neurons in PFC of different groups. Scale bar = 2 μm. (L) Ratio of damaged mitochondria (Representative TEM images were used to calculate the percentage of damaged mitochondria. Mitochondrial damage was defined as the presence of mitochondrial swelling and/or cristae disruption, as described in the Materials and Methods.) (**p < 0.01, ***p < 0.001. n = 5 mice per group). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: GLP-1R, Glucagon-like Peptide-1 receptor; CREB, cAMP Response Element-Binding; OXPHOS, Oxidative phosphorylation; Sema, Semaglutide; NDUFB8, Ubiquinone oxidoreductase subunit B8; SDHB, Succinate dehydrogenase complex iron sulfur subunit B; UQCRC2, Ubiquinol-cytochrome c reductase core protein 2; MTCO1, Mitochondrially encoded cytochrome c oxidase I; vATP5A, ATP synthase F1 subunit alpha.

Article Snippet: Immunofluorescence staining was performed using primary antibodies against GLP-1R ( GB113881 , 1:5000, Servicebio) and NeuN (GB11138, 1:5000, Servicebio).

Techniques: Expressing, Western Blot, Disruption, Binding Assay, Phospho-proteomics

Journal: Frontiers in Pharmacology

Article Title: Activation of GLP-1R ameliorates alcohol withdrawal induced anxiety-like behavior by regulating neuronal mitochondrial quality control

doi: 10.3389/fphar.2026.1820128

Figure Lengend Snippet: Regulation of mitochondrial quality control by Semaglutide in CEE mice. (A) Mitochondrial quality control, including fission, fusion, and mitophagy. (B–D) Protein expression of mitochondrial FIS1, DRP1 and p-DRP1 were detected (*p < 0.05, **p < 0.01, ***p < 0.001. n = 6 mice per group). (E–G) Protein expression of mitochondrial MFN1 and MFN2 were detected (*p < 0.05. n = 6 mice per group). (H–J) Protein expressions of mitochondrial Parkin and Pink1 were detected *p < 0.05, ***p < 0.001. n = 6 mice per group). (K-M) Protein expressions of LC3B and P62 were detected (*p < 0.05, **p < 0.01. n = 6 mice per group). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: GLP-1R, Glucagon-like Peptide-1 receptor; Sema, Semaglutide; DRP1DRP1, Dynamin-related protein-1; FIS1, Fission, mitochondrial 1; MFN1, Mitofusin 1; MFN2, Mitofusin 2; Pink1, PTEN-induced kinase 1; Parkin, Parkin RBR E3 ubiquitin-protein ligase; LC3, Microtubule-associated protein 1A/1B-light chain 3.

Article Snippet: Immunofluorescence staining was performed using primary antibodies against GLP-1R ( GB113881 , 1:5000, Servicebio) and NeuN (GB11138, 1:5000, Servicebio).

Techniques: Control, Expressing, Ubiquitin Proteomics

Journal: Frontiers in Pharmacology

Article Title: Activation of GLP-1R ameliorates alcohol withdrawal induced anxiety-like behavior by regulating neuronal mitochondrial quality control

doi: 10.3389/fphar.2026.1820128

Figure Lengend Snippet: Synaptic morphology in PFC pyramidal neurons of CEE mice treated with Semaglutide. (A-C) Protein expressions of PSD95 and SYN were detected (*p < 0.05, ***p < 0.001. n = 6 mice per group). (D) Representative images of neurons in PFC labeled using Golgi staining. Scale bar = 20 μm. (E) Sholl analysis of PFC neurons revealing alterations in basal dendritic intersections at distinct distances from the soma. Two-way ANOVA with repeated measures revealed a significant effect of drug treatment and distance from the soma (*p < 0.05, **p < 0.01, ***p < 0.001. n = 4 mice per group). (F,G) Total dendritic length and dendritic branch number (**p < 0.01. n = 4 mice per group). (H) Representative images of dendritic spines by Golgi staining. (I) Spine density. Scale bar = 10 μm. (***P < 0.001. n = 4 mice per group). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: GLP-1R, Glucagon-like Peptide-1 receptor; Sema, Semaglutide; PSD95, Postsynaptic density protein 95; SYN, Synuclein.

Article Snippet: Immunofluorescence staining was performed using primary antibodies against GLP-1R ( GB113881 , 1:5000, Servicebio) and NeuN (GB11138, 1:5000, Servicebio).

Techniques: Labeling, Staining

Journal: Comprehensive Physiology

Article Title: Pancreatic Islet Cell Crosstalk: Insight Into α‐/β‐Cell Compensatory Mechanisms

doi: 10.1002/cph4.70158

Figure Lengend Snippet: Quantification of glucagon/GLP‐1 linked transcripts/proteins. (A) Schema of paracrine communication of α‐/β‐cells through glucagon and GLP‐1. Created in Biorender. Plecita, L. (2026), https://BioRender.com/0mxd7q7 . (B) qPCR quantification of preproglucagon‐related transcripts in WT (gray) and Nox4 βKO (red) islets ( n = 4), T ‐test (Welch's correction), p = 0.0062, p = 0.4661, p = 0.0257, p = 0.0034, p = 0.9534. (C) Quantification of PC1/3 positive cells in WT (gray) and Nox4 βKO (red) islets ( n = 3–4), T ‐test, p = 0.0007; and (D) of PC1/3 positive α‐cells of WT (gray) and Nox4 βKO (red) islets (right panel) ( n = 4–6), T ‐test, p = 0.0005. (E) Quantification of GCGR relative protein levels in WT (gray) and Nox4 βKO (red) pancreatic lysates ( n = 7), T ‐test, p = 0.0202. Representative blots are presented (F) Quantification of GLP‐1R relative protein in WT (gray) and Nox4 βKO (red) pancreatic lysates ( n = 9), T ‐test, p = 0.0130. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Equal protein amounts were separated by SDS‐PAGE, transferred to PVDF, and probed for insulin (ab181547, Abcam, UK), GCGR (ab75240, Abcam, UK), and GLP‐1R (BS‐1559R, Bioss, ThermoFisher Scientific, USA).

Techniques:

Journal: Comprehensive Physiology

Article Title: Pancreatic Islet Cell Crosstalk: Insight Into α‐/β‐Cell Compensatory Mechanisms

doi: 10.1002/cph4.70158

Figure Lengend Snippet: Functional analysis of prediabetic islets while manipulating GCG/GLP‐1 signaling. (A) Quantification of insulin secretion of WT (gray) and Nox4 βKO (red) islets upon non‐stimulating (solid colors) or glucose‐stimulating (pattern colors) conditions ( n = 6), ANOVA, p < 0.0001, p < 0.0001, p = 0.8421. (B) Quantification of glucagon secretion of WT (gray) and Nox4 βKO (red) islets upon non‐stimulating (solid colors) or glucose‐stimulating (pattern colors) conditions ( n = 6–7), ANOVA, p = 0.0112, p = 0.0523, p < 0.0001. (C) Quantification of GLP‐1 of WT (gray) and Nox4 βKO (red) pancreases ( n = 11–12), T ‐test, p = 0.0018. (D) Analysis of insulin secretion of WT islets (gray labeling) and Nox4 βKO islets (red labeling) upon non‐stimulating condition (solid color), stimulation by glucose (crosshatch); glucose and GLP‐1 (stripes); glucose, GLP‐1 and GLP‐1R antagonist (Ex9) (diagonal grid); glucose, GLP‐1 and GCGR antagonist (Cro) (filled bricks); glucose, and GLP‐1R antagonist (Ex9) (light dots); glucose, and GCGR antagonist (Cro) (light bricks) and glucose; GLP‐1R antagonist (Ex9), and GCGR antagonist (Cro) (diagonal stripes), ( n = 3–7), ANOVA, for WT islets: p = 0.008, p = 0.0140, p = 0.0008, p = 0.7268, p = 0.0006, p = 0.9143, p = 0.0255; for Nox4 βKO islets: p = 0.9723, p = 0.0013, p = 0.4586, p = 0.0008, p = 0.4112, p = 0.2657, p = 0.7645. Only significant statistics for the relevant samples are presented. (E) Analysis of insulin secretion of WT islets (gray labeling) and Nox4 βKO islets (red labeling) upon non‐stimulating condition (solid color), stimulation by glucose (crosshatch); glucose and glucagon (stripes); glucose, glucagon and GCGR antagonist (Cro) (bricks); glucose, glucagon and GLP‐1R antagonist (Ex9) (light dots); ( n = 3–7), ANOVA, for WT islets: p = 0.0001, p = 0.0001, p < 0.0001, p = 0.0003 and for Nox4 βKO islets: p = 0.9990, p = 0.7679, p = 0.3741, p = 0.0315. Only significant statistics for the relevant samples are presented. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Equal protein amounts were separated by SDS‐PAGE, transferred to PVDF, and probed for insulin (ab181547, Abcam, UK), GCGR (ab75240, Abcam, UK), and GLP‐1R (BS‐1559R, Bioss, ThermoFisher Scientific, USA).

Techniques: Functional Assay, Labeling

Journal: Comprehensive Physiology

Article Title: Pancreatic Islet Cell Crosstalk: Insight Into α‐/β‐Cell Compensatory Mechanisms

doi: 10.1002/cph4.70158

Figure Lengend Snippet: Quantification of cAMP signaling in prediabetic islets. (A) cAMP quantification in WT islets (gray labeling) and Nox4 βKO islets (red labeling) upon non‐stimulating condition (solid color), stimulation by glucose (crosshatch); glucose and GLP‐1 (stripes); glucose, GLP‐1 and GLP‐1R antagonist (Ex9) (diagonal grid); glucose, GLP‐1 and GCGR antagonist (Cro) (filled bricks); glucose, and GLP‐1R antagonist (Ex9) (light dots); glucose, and GCGR antagonist (Cro) (light bricks) and glucose; GLP‐1R antagonist (Ex9), and GCGR antagonist (Cro) (diagonal stripes), ( n = 3–14), ANOVA, for WT islets: p = 0.0054, p = 0.0010, p < 0.0001, p = 0.0198, p = 0.6772, p = 0.9721, p = 0.8916; for Nox4 βKO islets: P = 0.0008, p = 0.0009, p < 0.0001, p = 0.9993, p = 0.8804, p = 0.7824. Only significant statistics for the relevant samples are presented. (B) cAMP quantification in WT islets (gray labeling) and Nox4 βKO islets (red labeling) upon non‐stimulating condition (solid color), stimulation by glucose (crosshatch); glucose and glucagon (stripes); glucose, glucagon and GCGR antagonist (Cro) (filled bricks); glucose, glucagon and GLP‐1R antagonist (Ex9) (light dots); ( n = 3–9), ANOVA, for WT islets: p = 0.0009, p < 0.0001, p < 0.0001, p = 0.0573; for Nox4 βKO islets: p = 0.9997, p = 0.8095, p = 0.9998, p = 0.0423. Only significant statistics for the relevant samples are presented. (C) Quantification of PDE transcripts in WT (gray) and Nox4 βKO (red) islets, ( n = 3), statistical significance was assessed using edgeR, p = 0.396, p = 0.000149. (D) Quantification of adenylate cyclase transcripts in WT (gray) and Nox4 βKO (red) islets, ( n = 3), statistical significance was assessed using edgeR, p = 0.333, p = 0.0254, p = 0.0263. (E) Quantification of PKA subunit (regulatory, catalytic) transcripts in WT (gray) and Nox4 βKO (red) islets, ( n = 3), statistical significance was assessed using edgeR, p = 0.0571, p = 0.000587, p = 0.307, p = 0.591. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Equal protein amounts were separated by SDS‐PAGE, transferred to PVDF, and probed for insulin (ab181547, Abcam, UK), GCGR (ab75240, Abcam, UK), and GLP‐1R (BS‐1559R, Bioss, ThermoFisher Scientific, USA).

Techniques: Labeling