scholarly journals Establishment of a conditional transgenic mouse model expressing human uncoupling protein 2 in vascular smooth muscle cells

2012 ◽  
Vol 4 (3) ◽  
pp. 545-547 ◽  
Author(s):  
SHUANGTAO MA ◽  
DE LI ◽  
DACHUN YANG ◽  
YAN TAN ◽  
BING TANG ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Transgenic Mouse ◽  
Vascular Smooth Muscle Cells ◽  
Uncoupling Protein ◽  
Transgenic Mouse Model ◽  
Uncoupling Protein 2 ◽  
Conditional Transgenic Mouse

scholarly journals Pressure-induced constriction is inhibited in a mouse model of reduced βENaC

2009 ◽  
Vol 297 (3) ◽  
pp. R723-R728 ◽  
Author(s):  
Lauren G. VanLandingham ◽  
Kimberly P. Gannon ◽  
Heather A. Drummond
Keyword(s):  
Smooth Muscle ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Cerebral Arteries ◽  
Muscle Cells ◽  
Normal Pressure ◽  
Quantitative Immunofluorescence ◽  
Mechanosensitive Ion Channel

Recent studies suggest certain epithelial Na+channel (ENaC) proteins may be components of mechanosensitive ion channel complexes in vascular smooth muscle cells that contribute to pressure-induced constriction in middle cerebral arteries (MCA). However, the role of a specific ENaC protein, βENaC, in pressure-induced constriction of MCAs has not been determined. The goal of this study was to determine whether pressure-induced constriction in the MCA is altered in a mouse model with reduced levels of βENaC. Using quantitative immunofluorescence, we found whole cell βENaC labeling in cerebral vascular smooth muscle cells (VSMCs) was suppressed 46% in βENaC homozygous mutant (m/m) mice compared with wild-type littermates (+/+). MCAs from βENaC +/+ and m/m mice were isolated and placed in a vessel chamber for myographic analysis. Arteries from βENaC+/+ mice constricted to stepwise increases in perfusion pressure and developed maximal tone of 10 ± 2% at 90 mmHg ( n = 5). In contrast, MCAs from βENaC m/m mice developed significantly less tone (4 ± 1% at 90 mmHg, n = 5). Vasoconstrictor responses to KCl (4–80 mM) were identical between genotypes and responses to phenylephrine (10−7-10−4M) were marginally altered, suggesting that reduced levels of VSMC βENaC specifically inhibit pressure-induced constriction. Our findings indicate βENaC is required for normal pressure-induced constriction in the MCA and provide further support for the hypothesis that βENaC proteins are components of a mechanosensor in VSMCs.

scholarly journals Phenotypic Alteration of Vascular Smooth Muscle Cells Precedes Elastolysis in a Mouse Model of Marfan Syndrome

2001 ◽  
Vol 88 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Tracie E. Bunton ◽  
Nancy Jensen Biery ◽  
Loretha Myers ◽  
Barbara Gayraud ◽  
Francesco Ramirez ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Marfan Syndrome ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Phenotypic Alteration

scholarly journals Deficiency in Aim2 affects viability and calcification of vascular smooth muscle cells from murine aortas and Angiotensin-II induced aortic aneurysms

2020 ◽  
Author(s):  
Markus Wortmann ◽  
Muhammad Arshad ◽  
Maani Hakimi ◽  
Dittmar Böckler ◽  
Susanne Dihlmann
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Angiotensin Ii ◽  
Aortic Aneurysm ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells

Abstract Background: Phenotypic transformation of vascular smooth muscle cells is a key element in vascular remodeling and aortic aneurysm growth. Previously, deletion of several inflammasome components decreased formation of aortic aneurysm (AA) in the Angiotensin II (AngII) -induced mouse model. We hypothesized that the inflammasome sensor Absent in melanoma 2 (Aim2) might affect the phenotype of vascular smooth muscle cells (VSMC), thereby reducing AA formation. Methods : Aim2-/- mice and wild-type (WT) C57Bl/6J mice were used as an animal model. VSMC were isolated from 6 months old mice and grown in vitro . Young (passage 3-5) and senescent (passage 7-12) cells were analyzed in vitro for calcification in mineralization medium by Alizarin Red S staining. Expression of calcification and inflammatory markers were studied by real-time RT-PCR and Western blotting, release of cytokines was determined by ELISA. To induce AA, osmotic mini-pumps loaded with AngII (1500 ng/kg bodyweight/min) were implanted for 28 days in male mice at 6 months of age. Results : Compared with VSMC from WT mice, VSMC isolated from Aim2-/- mice were larger, less viable, and underwent stronger calcification in mineralization medium, along with induction of Bmp4 and repression of Tnfsf11/Rankl gene expression. In addition, Aim2 deficiency was associated with reduced inflammasome gene expression and release of Interleukin-6. Using the mouse model of AngII induced AA, Aim2 deficiency reduced AA incidence to 48.4% (15/31) in Aim2-/- mice versus 76.5% (13/17) in WT mice. In contrast to Aim2-/- mice, AA from WT mice expressed significantly increased levels of alpha-smooth muscle actin/ Acta2 , indicating tissue remodeling. Reduced cell proliferation in Aim2-/- mice was indicated by significantly increased p16ink4a/ Cdkn2a expression in untreated and AngII-infused aortas, and by significantly lower amounts of proliferating (Ki67 positive) VSMC in AngII-infused Aim2-/- mice. Conclusions: Our results suggest a role for Aim2 in regulating VSMC proliferation and transition to an osteoblast-like or osteoclast-like phenotype, thereby modulating the response of VSMC in aortic remodeling and AA Formation.

scholarly journals Deficiency in Aim2 affects viability and calcification of vascular smooth muscle cells from murine aortas and Angiotensin-II induced aortic aneurysms

2020 ◽  
Author(s):  
Markus Wortmann ◽  
Muhammad Arshad ◽  
Maani Hakimi ◽  
Dittmar Böckler ◽  
Susanne Dihlmann
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Angiotensin Ii ◽  
Aortic Aneurysm ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells

Abstract Background: Phenotypic transformation of vascular smooth muscle cells is a key element in vascular remodeling and aortic aneurysm growth. Previously, deletion of several inflammasome components decreased formation of aortic aneurysm (AA) in the Angiotensin II (AngII) -induced mouse model. We hypothesized that the inflammasome sensor Absent in melanoma 2 (Aim2) might affect the phenotype of vascular smooth muscle cells (VSMC), thereby reducing AA formation. Methods: Aim2-/- mice and wild-type (WT) C57Bl/6J mice were used as an animal model. VSMC were isolated from 6 months old mice and grown in vitro. Young (passage 3-5) and senescent (passage 7-12) cells were analyzed in vitro for calcification in mineralization medium by Alizarin Red S staining. Expression of calcification and inflammatory markers were studied by real-time RT-PCR and Western blotting, release of cytokines was determined by ELISA. To induce AA, osmotic mini-pumps loaded with AngII (1500 ng/kg bodyweight/min) were implanted for 28 days in male mice at 6 months of age.Results: Compared with VSMC from WT mice, VSMC isolated from Aim2-/- mice were larger, less viable, and underwent stronger calcification in mineralization medium, along with induction of Bmp4 and repression of Tnfsf11/Rankl gene expression. In addition, Aim2 deficiency was associated with reduced inflammasome gene expression and release of Interleukin-6. Using the mouse model of AngII induced AA, Aim2 deficiency reduced AA incidence to 48.4% (15/31) in Aim2-/- mice versus 76.5% (13/17) in WT mice. In contrast to Aim2-/- mice, AA from WT mice expressed significantly increased levels of alpha-smooth muscle actin/Acta2, indicating tissue remodeling. Reduced cell proliferation in Aim2-/- mice was indicated by significantly increased p16ink4a/Cdkn2a expression in untreated and AngII-infused aortas, and by significantly lower amounts of proliferating (Ki67 positive) VSMC in AngII-infused Aim2-/- mice. Conclusions: Our results suggest a role for Aim2 in regulating VSMC proliferation and transition to an osteoblast-like or osteoclast-like phenotype, thereby modulating the response of VSMC in aortic remodeling and AA formation.

Abstract P221: Vascular Smooth Muscle-Specific Overexpression of Cyp4a12-20-HETE Synthase Causes Hypertension and Vascular Remodeling

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Rebecca Hutcheson ◽  
Frank Zhang ◽  
Katherine Gotlinger ◽  
Michal Schwartzman
Keyword(s):  
Blood Pressure ◽  
Smooth Muscle ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Transgenic Mouse ◽  
Vascular Remodeling ◽  
Transgenic Mouse Model ◽  
Vascular Effects ◽  
Cmv Promoter ◽  
Primary Mouse

20-Hydroxyeicosatetraenoic acid (20-HETE) is a potent vasoactive eicosanoid of the microcirculation exhibiting effects on vascular smooth muscle (VSM) function that include stimulation of contractility, migration and growth. We have previously demonstrated in mice which globally overexpress 20-HETE that hypertension as well as vascular remodeling is not fully prevented by pharmacological normalization of blood pressure. However, both hypertension and vascular remodeling are prevented by antagonism of 20-HETE, suggesting that 20-HETE exerts vascular effects independent of hypertension. To examine the contribution of VSM-derived 20-HETE to hypertension and vascular remodeling, we have developed a transgenic mouse model that overexpresses the primary mouse 20-HETE synthase, Cyp4a12, specifically in VSM by crossbreeding floxed Cyp4a12 with Myh11-Cre mice. All mice carry the floxed Cyp4a12 gene and express a GFP protein under control of a modified CMV promoter. Cre-recombination excises the GFP cDNA and inserts the Cyp4a12 cDNA. Three to four month old male Cyp4a12-floxed mice carrying the Myh11-Cre gene (Cyp4a12 fl/fl Myh11 Cre+/- ) exhibited higher systolic blood pressure than control mice Cyp4a12 fl/fl Myh11 Cr-/- (WT) mice (135 vs. 115 mmHg, n=4, *p<0.05). 20-HETE production in Cyp4a12 fl/fl Myh11 Cre+/- mice were higher than in WT in mesenteric (3333±891 vs. 545± 196 ng/mg/h) and renal interlobar arteries (RIA; 1859±376 vs. 242±62 ng/mg/h). Plasma levels of 20-HETE were also elevated (324±61 vs. 185±34 pg/mL) while urinary levels were not significantly different (146±26 vs. 117±29 pg/mL). We also observed increased medial thickness and decreased lumen area of blood vessels by Myh11 immunofluorescence in heart and kidney sections. RIA from male Myh11-Cyp4a12 mice displayed higher constrictor sensitivity to phenylephrine and impaired relaxation to acetylcholine compared to WT. Taken together, these data suggest that this model displays hypertension and pathological hypertrophic vascular remodeling. We therefore conclude that the Cyp4a12 fl/fl Myh11 Cre/- is a promising and unique transgenic mouse model to examine the contribution of smooth muscle-derived 20-HETE to hypertension and hypertrophic vascular remodeling.

scholarly journals Deficiency in Aim2 affects viability and calcification of vascular smooth muscle cells from murine aortas and angiotensin-II induced aortic aneurysms

2020 ◽  
Vol 26 (1) ◽  
Author(s):  
Markus Wortmann ◽  
Muhammad Arshad ◽  
Maani Hakimi ◽  
Dittmar Böckler ◽  
Susanne Dihlmann
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Angiotensin Ii ◽  
Aortic Aneurysm ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells

Abstract Background Phenotypic transformation of vascular smooth muscle cells is a key element in vascular remodeling and aortic aneurysm growth. Previously, deletion of several inflammasome components decreased formation of aortic aneurysm (AA) in the Angiotensin II (AngII) -induced mouse model. We hypothesized that the inflammasome sensor Absent in melanoma 2 (Aim2) might affect the phenotype of vascular smooth muscle cells (VSMC), thereby reducing AA formation. Methods Aim2−/− mice and wild-type (WT) C57Bl/6 J mice were used as an animal model. VSMC were isolated from 6 months old mice and grown in vitro. Young (passage 3–5) and senescent (passage 7–12) cells were analyzed in vitro for calcification in mineralization medium by Alizarin Red S staining. Expression of calcification and inflammatory markers were studied by real-time RT-PCR and Western blotting, release of cytokines was determined by ELISA. To induce AA, osmotic mini-pumps loaded with AngII (1500 ng/kg bodyweight/min) were implanted for 28 days in male mice at 6 months of age. Results Compared with VSMC from WT mice, VSMC isolated from Aim2−/− mice were larger, less viable, and underwent stronger calcification in mineralization medium, along with induction of Bmp4 and repression of Tnfsf11/Rankl gene expression. In addition, Aim2 deficiency was associated with reduced inflammasome gene expression and release of Interleukin-6. Using the mouse model of AngII induced AA, Aim2 deficiency reduced AA incidence to 48.4% (15/31) in Aim2−/− mice versus 76.5% (13/17) in WT mice. In contrast to Aim2−/− mice, AA from WT mice expressed significantly increased levels of alpha-smooth muscle actin/Acta2, indicating tissue remodeling. Reduced cell proliferation in Aim2−/− mice was indicated by significantly increased p16ink4a/Cdkn2a expression in untreated and AngII-infused aortas, and by significantly lower amounts of proliferating (Ki67 positive) VSMC in AngII-infused Aim2−/− mice. Conclusions Our results suggest a role for Aim2 in regulating VSMC proliferation and transition to an osteoblast-like or osteoclast-like phenotype, thereby modulating the response of VSMC in aortic remodeling and AA formation.

scholarly journals Parathyroid hormone induces transition of myofibroblasts in arteriovenous fistula and increases maturation failure

Endocrinology ◽  
2021 ◽  
Author(s):  
Chung-Te Liu ◽  
Shih-Chang Hsu ◽  
Hui-Ling Hsieh ◽  
Cheng-Hsien Chen ◽  
Chun-You Chen ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Parathyroid Hormone ◽  
Mouse Model ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Secondary Hyperparathyroidism ◽  
Arteriovenous Fistula ◽  
Vascular Smooth Muscle Cells ◽  
Cell Model ◽  
Muscle Cells

Abstract Purpose Arteriovenous fistula (AVF) maturation failure remains a clinical dilemma and its pathobiology is largely unclear. Secondary hyperparathyroidism is a complication of chronic renal failure that associated with cardiovascular disease. While parathyroid hormone (PTH) has prosclerotic effect on vascular smooth muscle cells, its role on AVF maturation failure was unknown. Methods Patients receiving AVF creation were enrolled retrospectively to investigate the association between plasma PTH and AVF maturation. A mouse model of secondary hyperparathyroidism and aortocaval AVF was used to investigate the effect of PTH on AVF lesion. A cell model of vascular smooth muscle cell treated with PTH in pressurized culture system was used to disclose the signaling pathway underlying the effect of PTH on AVF lesion. Results In patients receiving AVF creation, higher PTH was associated with increased risk for maturation failure. In mouse model, vascular wall thickness and myofibroblasts of AVF significantly increased with higher PTH. When the same mice was treated by cinacalcet, AVF lesions were attenuated by suppression of PTH. Cell model showed that PTH increased the marker of myofibroblasts, integrin β6 subunit (ITGB6) via the phospho-Akt pathway. Finally, in the same model of mice AVF, higher PTH also increased the expression of ITGB6 in the smooth muscle layer of AVF, suggesting the transition to myofibroblast. Conclusions Overall, our results suggest that higher PTH increased the risk of AVF maturation failure through increasing the transition of vascular smooth muscle cells to myofibroblasts. Lowering PTH may be a strategy to enhance AVF maturation.

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