Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A – C ) Body weight and oral glucose tolerance test with calculated area under the glucose curve in control Glp1r Tie2+/+ and Glp1r Tie2–/– male mice with acute semaglutide (10 μg/kg) treatment 120 minutes before glucose challenge ( n = 7–13). ( D – F ) Quantification of gene expression relative to Rpl32 within the kidney, renal artery, and lung in Glp1r Tie2+/+ and Glp1r Tie2–/– male mice ( n = 4–6). ( G – I ) Systolic blood pressure, diastolic blood pressure, and mean arterial pressure (MAP) in control and Glp1r Tie2–/– male mice treated acutely with semaglutide 120 minutes prior to BP measurements ( n = 5-14). ( J – O ) BP measurements following ( J – L ) induction of hypertension with Ang II for 2 weeks and ( M – O ) acute semaglutide treatment in angiotensin II-induced hypertensive mice ( n = 5–7). Data are presented as mean ± SD. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001 by unpaired t test ( A ), or 2-way ANOVA followed by Tukey post hoc tests ( C – O ). BP, blood pressure; Sema, semaglutide; Veh, vehicle; Ang, angiotensin.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control, Gene Expression

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A – C ) Annotation maps of the human kidney single-cell transcriptome , showing expression of GLP1R , and vascular smooth muscle marker α1A adrenergic receptor ( ADRA1A ). ( D – F ) An annotation map of the mouse kidney single-cell transcriptome , showing expression of Glp1r and VSMC marker myosin heavy chain 11 ( Myh11 ). ( G – L ) Quantification of Glp1r expression relative to Rpl32 within the ( G ) kidney, ( H ) renal artery, ( I ) lung, ( J ) aortic arch, ( K ) abdominal aorta, and ( L ) carotid artery in control ( Glp1r VSM+/+ ) and Glp1r VSM–/– male mice (n = 5–14). ( M ) Double immunofluorescence staining of α-smooth muscle actin (left panel), GLP-1R (middle panel) and both GLP-1R and α-smooth muscle actin (right panel) in Glp1r VSM+/+ and Glp1r VSM–/– kidney. ( N ) Average food intake per mouse within each cage of group housed mice over 24 hours with or without acute treatment with semaglutide in Glp1r VSM+/+ and Glp1r VSM–/– mice ( n = 4–6). ( O – Q ) Body weight and oral glucose tolerance with calculated area under the curve in Glp1r VSM+/+ and Glp1r VSM–/– mice following acute semaglutide (10 μg/kg) treatment 120 minutes before oral glucose challenge (time 0) ( n = 18–23). Data are presented as mean ± SD. *** P ≤ 0.001 and **** P ≤ 0.0001 by unpaired t test ( G – L and O ), or 2-way ANOVA followed by Tukey post hoc tests ( N and Q ). Sema, semaglutide; Veh, vehicle. Original magnification, ×31.5.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Single Cell, Expressing, Marker, Control, Double Immunofluorescence Staining

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A ) Systolic blood pressure, ( B ) diastolic blood pressure, and ( C ) mean arterial pressure (MAP) in control Glp1r VSM+/+ and Glp1r VSM–/– normotensive mice ( n = 10–14). ( D – F ) Glp1r VSM+/+ and Glp1r VSM–/– mice were treated acutely with semaglutide (10 μg/kg) 2 hours prior to BP measurements ( n = 12). ( G – X ) BP measurements before (control) or after ( G – I ) induction of hypertension with 2 weeks of access to L-NAME in drinking water and ( J – L ) acute semaglutide treatment in L-NAME-induced hypertensive mice ( n = 17–18), ( M – O ) induction of hypertension 3 weeks after AAV8- Ren1d viral transfection ( n = 5–7) and ( P – R ) acute semaglutide treatment in AAV8-Ren1d-induced hypertensive mice ( n = 8-13), and ( S – U ) induction of hypertension 3 weeks after Ang II minipump implantation ( n = 3-7) and ( V – X ) acute semaglutide treatment in Ang II–induced hypertensive mice ( n = 6-7). Data are presented as mean ± SD. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001 by unpaired t test ( A – C ), or 2-way ANOVA followed by Tukey post hoc tests ( D – X ). BP, blood pressure; Sema, semaglutide; Veh, vehicle; Ang, angiotensin.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control, Transfection

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A – C ) Body weight and oral glucose tolerance test with calculated area under the glucose curve in control Glp1r Tie2+/+ and Glp1r Tie2–/– male mice with acute semaglutide (10 μg/kg) treatment 120 minutes before glucose challenge ( n = 7–13). ( D – F ) Quantification of gene expression relative to Rpl32 within the kidney, renal artery, and lung in Glp1r Tie2+/+ and Glp1r Tie2–/– male mice ( n = 4–6). ( G – I ) Systolic blood pressure, diastolic blood pressure, and mean arterial pressure (MAP) in control and Glp1r Tie2–/– male mice treated acutely with semaglutide 120 minutes prior to BP measurements ( n = 5-14). ( J – O ) BP measurements following ( J – L ) induction of hypertension with Ang II for 2 weeks and ( M – O ) acute semaglutide treatment in angiotensin II-induced hypertensive mice ( n = 5–7). Data are presented as mean ± SD. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001 by unpaired t test ( A ), or 2-way ANOVA followed by Tukey post hoc tests ( C – O ). BP, blood pressure; Sema, semaglutide; Veh, vehicle; Ang, angiotensin.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control, Gene Expression

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A – D ) Urine volume, urine sodium, urine chloride, and urine protein in normotensive control Glp1r VSM+/+ and Glp1r VSM–/– male mice treated acutely with semaglutide (10 μg/kg) 60 minutes prior to water gavage; urine was collected 180 minutes following gavage ( n = 15–22). ( E ) Glomerular filtration rate was measured in Glp1r VSM+/+ and Glp1r VSM–/– mice treated acutely with semaglutide (10 μg/kg) 30 minutes before a bolus of FITC- sinistrin ( n = 5–10). ( F ) Myography readings depicting vasorelaxation of ex vivo mesenteric arteries, assessed after phenylephrine preconstriction (1 μM), in response to semaglutide (10–1,000 nM). ( G – J ) Two weeks after L-NAME–induced hypertension, mice were treated acutely with semaglutide (i.p.) 60 minutes prior to a water gavage, and urine was collected 180 minutes later. ( G – J ) Urine volume, urine sodium, urine chloride, and urine protein were measured ( n = 18–22). Urine sodium, chloride, and protein levels were normalized to creatinine concentration. Data are presented as mean ± SD. * P ≤ 0.05, *** P ≤ 0.001, and **** P ≤ 0.0001 by 2-way ANOVA followed by Tukey post hoc-tests ( A – E and G – J ) or multiple unpaired t tests ( F ). Sema, semaglutide; Veh vehicle.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control, Filtration, Ex Vivo, Concentration Assay

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A ) Volcano plot depicting changes in the distribution of significance and fold change of identified proteins in renal artery between Glp1r VSM+/+ and Glp1r VSM–/– mice. ( B and C ) Feature plots for identified Reactome pathways ( B ) positively and ( C ) negatively correlated with VSMC Glp1r knockdown. The color scheme is based on p-value distribution. ( D ) Volcano plot depicting changes in quantified protein between Glp1r VSM+/+ and Glp1r VSM–/– mice in the kidney. ( E and F ) Feature plots for identified Reactome pathways ( E ) positively and ( F ) negatively correlated with VSMC Glp1r KO. Statistical comparisons were made using unpaired t test between groups.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Knockdown

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: ( A ) Mice were treated with semaglutide (10 μg/kg) 4 hours before renal artery excision and processing. ( A ) Volcano plot depicting changes in the distribution of significance and fold change of identified proteins between control mice treated with vehicle (Veh) and semaglutide (Sema). ( B and C ) Feature plots for identified Reactome pathways ( B ) positively and ( C ) negatively correlated with vehicle versus semaglutide treatment. The color scheme is based on P value distribution. ( D ) Volcano plot depicting changes in quantified protein between Glp1r VSM–/– mice treated with vehicle or semaglutide; highlighted proteins represent significantly changed targets in control mice. ( E ) Venn diagram depicting overlap of proteins regulated within the separate comparisons. ( F and G ) Heatmaps comparing proteins in control and Glp1r VSM–/– mice that are ( F ) positively and ( G ) negatively correlated with semaglutide treatment. Statistical comparisons were made using unpaired t test between groups.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control

Journal: JCI Insight

Article Title: Semaglutide reduces murine blood pressure through the vascular smooth muscle GLP-1 receptor

doi: 10.1172/jci.insight.201148

Figure Lengend Snippet: Mice were treated with semaglutide (10 mg/kg) 4 hours before kidney excision and processing. ( A ) Volcano plot depicting changes in the distribution of significance and fold change of identified proteins between control mice treated with vehicle (Veh) and semaglutide (Sema). ( B and C ) Feature plots for identified Reactome pathways ( B ) positively and ( C ) negatively correlated with vehicle versus semaglutide treatment. The color scheme is based on P value distribution. ( D ) Volcano plot depicting changes in quantified protein between Glp1r VSM–/– mice treated with vehicle or semaglutide; highlighted proteins represent significantly changed targets in control mice. ( E ) Venn diagram depicting overlap of proteins regulated within the separate comparisons. ( F and G ) Heatmaps comparing proteins in control and Glp1r VSM–/– mice that are ( F ) positively and ( G ) negatively correlated with semaglutide treatment. Statistical comparisons were made using unpaired t test between groups.

Article Snippet: Glp1r Tie2–/– mice were generated by crossing Glp1r fl/fl mice with Tg (Tek-Cre)1Ywa ( Tie2) Cre mice (Jackson Laboratory, 008863) to target endothelial and immune cells in the Tie2 domain, as previously described ( , ).

Techniques: Control

Journal: The Journal of Clinical Investigation

Article Title: Lysophosphatidic acid–induced YAP/TAZ activation promotes developmental angiogenesis by repressing Notch ligand Dll4

doi: 10.1172/JCI121955

Figure Lengend Snippet: (A) LPA (10 μM, 3 hours) affected mRNA expression of various angiogenesis-related genes in serum-starved HUVECs. (B and C) LPA (10 μM) reduced mRNA (B) and protein (C) expression of DLL4 in a time-dependent manner in serum-starved HUVECs. (D) LPA4 and/or LPA6 siRNAs increased mRNA expression of DLL4. (E) LPA4/LPA6 and Gα12/Gα13 siRNAs increased protein expression of DLL4. (F) LPA4/LPA6 siRNAs increased protein expression of DLL4 and abolished the suppressive effect of LPA (10 μM, 3 hours) in serum-starved HUVECs. (G) Y27632 (10 μM, 10 minutes pretreatment) but not Ki16425 (10 μM, 10 minutes pretreatment) increased mRNA expression of DLL4 and abolished the suppressive effect of LPA (10 μM, 3 hours) in serum-starved HUVECs. (H) Rho inhibitor I (0.5 μg/mL, 6 hours pretreatment) and Y27632 (10 μM, 10 minutes pretreatment) increased protein expression of DLL4 and abolished the suppressive effect of LPA (10 μM, 3 hours) in serum-starved HUVECs. (I and J) Mouse lung ECs from Lpa4fl/Y;Lpa6fl/fl;Tie2-Cre (Lpa4;Lpa6ΔEC) mice displayed increased Dll4 (I) and Hey1 (J) mRNA expression. (K) Scheme for how LPA–LPA4/LPA6–Gα12/Gα13–Rho–ROCK signaling suppresses DLL4 expression. Data are mean ± SEM of triplicates. *P < 0.05, **P < 0.01, ***P < 0.001, 1-way ANOVA followed by Dunnett’s (B and D) or Tukey’s multiple-comparisons test (G) or 2-tailed unpaired Student’s t test (I and J). HUVECs were serum-starved for 8 hours or transfected with siRNAs for 96 hours. Unprocessed original scans of Western blots are shown in Supplemental Figure 12.

Article Snippet: Tie2 - Cre transgenic mice ( 75 ) were provided by RIKEN BioResource Research Center.

Techniques: Expressing, Transfection, Western Blot