title: Uncoupling Caveolae From Intracellular Signaling In Vivo.
abstract: BACKGROUND
  Caveolin-1 (Cav-1) negatively regulates endothelial nitric oxide (NO) synthase-derived NO production, and this has been mapped to several residues on Cav-1, including F92. Herein, we reasoned that endothelial expression of an F92ACav-1 transgene would let us decipher the mechanisms and relationships between caveolae structure and intracellular signaling.
  
  
  OBJECTIVE
  This study was designed to separate caveolae formation from its downstream signaling effects.
  
  
  RESULTS
  An endothelial-specific doxycycline-regulated mouse model for the expression of Cav-1-F92A was developed. Blood pressure by telemetry and nitric oxide bioavailability by electron paramagnetic resonance and phosphorylation of vasodilator-stimulated phosphoprotein were determined. Caveolae integrity in the presence of Cav-1-F92A was measured by stabilization of caveolin-2, sucrose gradient, and electron microscopy. Histological analysis of heart and lung, echocardiography, and signaling were performed.
  
  
  CONCLUSIONS
  This study shows that mutant Cav-1-F92A forms caveolae structures similar to WT but leads to increases in NO bioavailability in vivo, thereby demonstrating that caveolae formation and downstream signaling events occur through independent mechanisms.
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Jan R
authors.initials: JR
authors.lastname: Kraehling
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Hao
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Monica Y
authors.initials: MY
authors.lastname: Lee
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: David J
authors.initials: DJ
authors.lastname: Vinyard
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Heino
authors.initials: H
authors.lastname: Velazquez
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Xinran
authors.initials: X
authors.lastname: Liu
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Radu V
authors.initials: RV
authors.lastname: Stan
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: Gary W
authors.initials: GW
authors.lastname: Brudvig
authors.affiliation: From the Vascular Biology and Therapeutics Program (J.R.K., Z.H., M.Y.L., W.C.S.) and Department of Pharmacology (J.R.K., Z.H., M.Y.L., W.C.S.), Yale University School of Medicine, New Haven, CT; Department of Chemistry, Yale University, New Haven, CT (D.J.V., G.W.B.); Department of Internal Medicine, VA Connecticut Healthcare System, West Haven, CT (H.V.); Department of Cell Biology, Yale University, School of Medicine, New Haven, CT (X.L.); and Department of Pathology, Dartmouth Medical School, Lebanon, NH (R.V.S.).
authors.forename: William C
authors.initials: WC
authors.lastname: Sessa
mesh_terms: D000818:Animals; D001794:Blood Pressure / Q000187:drug effects / Q000502:physiology; D021941:Caveolae / Q000187:drug effects / Q000378:metabolism*; D051242:Caveolin 1 / Q000096:biosynthesis* / Q000235:genetics*; D004318:Doxycycline / Q000494:pharmacology; D006801:Humans; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009569:Nitric Oxide / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: C495541:Cav1 protein, mouse; D051242:Caveolin 1; D014475:Uncoupling Agents; D009569:Nitric Oxide; D004318:Doxycycline
keywords: caveolin-1; cell endothelial cell; eNOS; mice; nitric oxide; vascular function