Saltar al contenido
MilliporeSigma

Endothelial EphB4 maintains vascular integrity and transport function in adult heart.

eLife (2019-11-30)
Guillermo Luxán, Jonas Stewen, Noelia Díaz, Katsuhiro Kato, Sathish K Maney, Anusha Aravamudhan, Frank Berkenfeld, Nina Nagelmann, Hannes Ca Drexler, Dagmar Zeuschner, Cornelius Faber, Hermann Schillers, Sven Hermann, John Wiseman, Juan M Vaquerizas, Mara E Pitulescu, Ralf H Adams
RESUMEN

The homeostasis of heart and other organs relies on the appropriate provision of nutrients and functional specialization of the local vasculature. Here, we have used mouse genetics, imaging and cell biology approaches to investigate how homeostasis in the adult heart is controlled by endothelial EphB4 and its ligand ephrin-B2, which are known regulators of vascular morphogenesis and arteriovenous differentiation during development. We show that inducible and endothelial cell-specific inactivation of Ephb4 in adult mice is compatible with survival, but leads to rupturing of cardiac capillaries, cardiomyocyte hypertrophy, and pathological cardiac remodeling. In contrast, EphB4 is not required for integrity and homeostasis of capillaries in skeletal muscle. Our analysis of mutant mice and cultured endothelial cells shows that EphB4 controls the function of caveolae, cell-cell adhesion under mechanical stress and lipid transport. We propose that EphB4 maintains critical functional properties of the adult cardiac vasculature and thereby prevents dilated cardiomyopathy-like defects.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Insulina solution human, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Sacarosa, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Disolución de tripsina-EDTA, 1 ×, sterile; sterile-filtered, BioReagent, suitable for cell culture, 0.5 g porcine trypsin and 0.2 g EDTA, 4Na per liter of Hanks′ Balanced Salt Solution with phenol red
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
(Z)-4-Hydroxytamoxifen, ≥98% Z isomer
Sigma-Aldrich
Gelatina from porcine skin, powder, gel strength ~300 g Bloom, Type A, BioReagent, suitable for electrophoresis, suitable for cell culture
Roche
Kit de detección de muerte celular in situ,TMR rojo, sufficient for ≤50 tests
Sigma-Aldrich
Anti-vinculina monoclonal antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
Anticuerpo anti-actina, αmúsculo liso- Cy3 monoclonal de ratón, clone 1A4, purified from hybridoma cell culture
Sigma-Aldrich
HEPES solution, BioPerformance Certified, 1 M, suitable for cell culture, 0.2 μm filtered
Sigma-Aldrich
Mayer′s Hematoxylin Solution
Sigma-Aldrich
Seroalbúmina bovina solution, 30% in saline, fatty acid free, aseptically filled
Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
Oil Red O, certified by the Biological Stain Commission
Sigma-Aldrich
Polyvinylpyrrolidone, for molecular biology, nucleic acid hybridization tested, mol wt 360,000
Sigma-Aldrich
PP2, ≥98% (HPLC)