Myogenic vasoconstriction requires G12/G13 and LARG to maintain local and systemic vascular resistance
- Max Planck Institute for Heart and Lung Research, Germany
- University of Heidelberg, Germany
- UMR CNRS 6015 - INSERM 1083, Université d'Angers, France
- University of Regensburg, Germany
- JW Goethe University Frankfurt, Germany
- German Center for Cardiovascular Research (DZHK), Germany
Figures
Figure 1 with 2 supplements
Role of G12/G13, ARHGEF12 (LARG) and RhoA signaling in myogenic contraction.
(A–D) First order mesenteric arteries (A,B) and posterior cerebral arteries (C,D) were isolated from control (black circles), Sm-q/11-KO (blue circles) and Sm-12/13-KO (red circles) mice and were …
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Figure 1—source data 1
Analysis of pressure-induced changes in myogenic tone, RhoA-activation, Rock inhibition and intracellular calcium release in smooth muscle-specific Gαq/Gα111, Gα12/Gα13, and ARHGEF12 (LARG) deficient mice.
- https://doi.org/10.7554/eLife.49374.005
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Figure 1—source data 2
Western blot pictures (uncut) of pressure-inducedP-MYPT1 phosphorylation in smooth muscle-specificGα12/Gα13 deficient mice.
- https://doi.org/10.7554/eLife.49374.006
Figure 1—figure supplement 1
Effect of vascular smooth muscle-specific Gαq/Gα11, Gα12/Gα13, and ARHGEF12 (LARG) deficiency on myogenic tone responses.
(A-D) Representative traces of first order mesenteric arteries isolated from control (A, black), Sm-q/11-KO (B, blue), Sm-12/13-KO (C, red) and Sm-Larg-KO (D, green) mice showing changes in the …
Figure 1—figure supplement 2
Effect of L- type and TRP channel inhibitors on myogenic tone.
Determination of intracellular [Ca2+] during myogenic tone induction in first order mesenteric arteries incubated for 30 min in the presence (red circles) or absence (black circles) of L-type …
Figure 2 with 1 supplement
Effect of vascular smooth muscle-specific Gα12/Gα13 deficiency on vascular resistance and perfusion in peripheral organs.
(A) Flow-induced increase in vascular resistance in isolated perfused kidneys of control (black circles), Sm-12/13-KO (red circles) and Sm-q/11-KO mice (blues circles) (n = 12 mice (control) and n = …
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Figure 2—source data 1
Analysis of kidney, hind limb and brain perfusion in smooth muscle-specific Gαq/Gα11, Gα12/Gα13, and ARHGEF12 (LARG) deficient mice.
- https://doi.org/10.7554/eLife.49374.009
Figure 2—figure supplement 1
Effect of vascular smooth muscle-specific Gα12/Gα13 deficiency on Kidney and brain function.
(A–D) Urine volume. (A), glomerular filtration rate (GFR) (B), blood urea nitrogen (BUN) (C) and plasma creatinine levels (D) in wild-type and the indicated mutant mice (n = 7 mice (control and …
Figure 3 with 1 supplement
Cardiac structure and function of smooth muscle specific Gα12/Gα13-deficient mice.
(A) Hearts of wild-type (control, left) and Sm-12/13-KO mice (right) were stained 4 weeks after induction with hematoxylin and eosin (H and E), wheat germ agglutinin-AF488 (WGA-AF488), picrosirius …
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Figure 3—source data 1
Histological and MRI analysis of hypertrophy in smooth muscle-specific Gα12/Gα13deficient mice.
- https://doi.org/10.7554/eLife.49374.012
Figure 3—figure supplement 1
Hearts of wild-type (control) and Sm-12/13-KO mice were stained 34 weeks after tamoxifen induction with hematoxylin and eosin (H and E), as well as picrosirius red.
Scale bar: 5 mm.
Figure 4
Cardiac gene expression and systemic levels of mediators in Sm-12/13-KO mice.
(A and C) relative expression of myocardial genes and fibrosis marker genes in the hearts of control (white bars) and Sm-12/13-KO mice (red bars) 4 weeks (A) and 35 weeks (C) after tamoxifen …
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Figure 4—source data 1
Analysis of hypertrophy associated genes, catecholamines, aldosterone and angiotensin in smooth muscle-specific Gα12/Gα13deficient mice.
- https://doi.org/10.7554/eLife.49374.014
Figure 5
Effect of vascular smooth muscle-specific Gα12/Gα13 and LARG deficiency on LPS-induced hypotension.
(A and B), telemetric blood pressure measurements (systolic in A, diastolic in B) were performed in wild-type (control), Sm-12/13-KO and Sm-Larg-KO mice. At the indicated time point animals were …
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Figure 5—source data 1
Effects of LPS on blood pressure in Gα12/Gα13and ARHGEF12 (LARG) deficient mice.
- https://doi.org/10.7554/eLife.49374.016
Figure 6
Model of the role of G12/G13 in vascular myogenic tone.
VDCC, voltage-dependent Ca2+ channels; TRP, transient receptor potential channels; PLC-β, phospholipase C-β; PIP2, phosphatidyl inositol bisphosphate; DAG, diacyl glycerol, IP3, …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | Gna12−/− | PMID: 12077299 | RRID:MGI:3819345 | |
| Genetic reagent (Mus musculus) | Gna13flox/flox | PMID: 14528298 | RRID:MGI:3819345 | |
| Genetic reagent (Mus musculus) | Gnaqflox/flox | PMID: 11689889 | RRID:MGI:3819271 | |
| Genetic reagent (Mus musculus) | Gna11−/− | PMID: 9687499 | RRID:MGI:3819271 | |
| Genetic reagent (Mus musculus) | Largflox/flox | PMID: 18084302 | RRID:MGI:3819344 | |
| Genetic reagent (Mus musculus) | Myh11-CreERT2 | PMID: 18084302 | RRID:IMSR_JAX:019079 | |
| Antibody | anti-phospho-MYPT1 (rabbit polyclonal) | Merck Millipore | Catalog: 36–003 RRID:AB_310812 | WB (1:1000) |
| Antibody | anti-MYPT1 (rabbit polyclonal) | Cell Signalling Technologies | Catalog: 2634 RRID:AB_915965 | WB (1:500) |
| Antibody | anti-GAPDH (rabbit monoclonal) | Cell Signalling Technologies | Catalog: 2118 RRID:AB_561053 | WB (1:1000) |
| Commercialassay or kit | Rho-GLISA | Cytoskeleton Inc | BK124 | for determination of Rho activity |
| Ccommercial assay or kit | 3-CAT ELISA-ImmuSmol | LDN | BA E-5600 | for determination of adrenaline, dopamine and noradrenaline levels |
| Commercial assay or kit | Angiotensin II ELISA kit | ENZO Life Sciences | ADI-900–204 | |
| Commercial assay or kit | Aldosterone ELISA kit | ENZO Life Sciences | ADI-900–173 | |
| Chemical compound, drug | Y-27632 | Sigma Aldrich | Y0503 | 10 µM |
| Chemical compound, drug | 2-Aminoethyl diphenylborinate (2-ABP) | Sigma Aldrich | D9754 | 1 µM |
| Chemical compound, drug | Nifedipine | Sigma Aldrich | N7634 | 1 µM |
| Chemical compound, drug | Fura-2AM | Thermo Fisher Scientific | F1225 | 12.5 µM |
| Chemical compound, drug | Ionomycin | Sigma Aldrich | I0634 | 1 µM |
| Chemical compound, drug | LPS (Lipopolysaccharide) | Sigma Aldrich | L2630 | 10 mg/kg b.w |
| Other | WGA-AF488 | Thermo Fisher Scientific | W11261 | Histology (20 µg/ml) |
| Other | IB4-AF488 | Thermo Fisher Scientific | I21411 | Histology (1:200) |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.49374.018
