Vascular Hypoxic Preconditioning Relies on TRPV4-Dependent Calcium Influx and Proper Intercellular Gap Junctions Communication

Objective—We investigated the impact of hypoxia-reoxygenation on endothelial relaxation and aimed to clarify the role of transient receptor potential cation channels V4 (TRPV4) and gap junctions in the protective effect associated with hypoxic preconditioning on the vascular function. Methods and Results—By mimicking ischemia-reperfusion in C57BL/6 male mice in vivo, we documented a reduced NO-mediated relaxation and an increased endothelium-derived hyperpolarization (EDH[F])-mediated relaxation. Hypoxic preconditioning, however, restored NO relaxation and further improved the EDH(F) response. We also examined specifically 2 major effectors of the EDH(F) pathway, transient receptor potential cation channels V4 and connexins. We found that in endothelial cells, expression and activity of transient receptor potential cation channels V4 were increased by hypoxic stimuli independently of preconditioning which was interestingly associated with an increase of structural caveolar component caveolin-1 at membrane locations. Gap junctions, however, seemed to directly support EDH(F)-driven preconditioning as connexin 40 and connexin 43 expression increased and as in vivo carbenoxolone treatment completely inhibited the EDH(F) pathway and significantly reduced the protection afforded by preconditioning for the concomitant NO-mediated relaxation. Conclusion—Our work provides evidence on how transient receptor potential cation channels V4 and connexins might participate in preserving vasorelaxation under hypoxia and restoring the NO-mediated pathway in hypoxic preconditioning conditions pointing out caveolae as a common signaling location.

[1]  M. Billaud,et al.  Interaction between nitric oxide signaling and gap junctions: effects on vascular function. , 2012, Biochimica et biophysica acta.

[2]  R. Adapala,et al.  PKCα mediates acetylcholine-induced activation of TRPV4-dependent calcium influx in endothelial cells. , 2011, American journal of physiology. Heart and circulatory physiology.

[3]  I. Ambudkar,et al.  The dynamic complexity of the TRPC1 channelosome , 2011, Channels.

[4]  A. Zarbock,et al.  Spontaneous lung dysfunction and fibrosis in mice lacking connexin 40 and endothelial cell connexin 43. , 2011, The American journal of pathology.

[5]  C. de Wit,et al.  Distinct Endothelium-Derived Hyperpolarizing Factors Emerge In Vitro and In Vivo and Are Mediated in Part via Connexin 40–Dependent Myoendothelial Coupling , 2011, Hypertension.

[6]  J. Haefliger,et al.  Loss of connexin40 is associated with decreased endothelium-dependent relaxations and eNOS levels in the mouse aorta. , 2010, American journal of physiology. Heart and circulatory physiology.

[7]  P. Sorgen,et al.  Gap Junction Protein Cx37 Interacts With Endothelial Nitric Oxide Synthase in Endothelial Cells , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[8]  J. Balligand,et al.  The regulation of endothelial nitric oxide synthase by caveolin: a paradigm validated in vivo and shared by the ‘endothelium-derived hyperpolarizing factor’ , 2010, Pflügers Archiv - European Journal of Physiology.

[9]  Juan Fang,et al.  TRPV4-mediated endothelial Ca2+ influx and vasodilation in response to shear stress. , 2010, American journal of physiology. Heart and circulatory physiology.

[10]  A. Daiber,et al.  Tolerance to nitroglycerin-induced preconditioning of the endothelium: a human in vivo study. , 2010, American journal of physiology. Heart and circulatory physiology.

[11]  J. Balligand,et al.  eNOS activation by physical forces: from short-term regulation of contraction to chronic remodeling of cardiovascular tissues. , 2009, Physiological reviews.

[12]  B. Duling,et al.  Gap junctions in the control of vascular function. , 2009, Antioxidants & redox signaling.

[13]  B. Nilius,et al.  Role of cytochrome P450-dependent transient receptor potential V4 activation in flow-induced vasodilatation. , 2008, Cardiovascular research.

[14]  A. Pries,et al.  Negative-Feedback Loop Attenuates Hydrostatic Lung Edema via a cGMP-Dependent Regulation of Transient Receptor Potential Vanilloid 4 , 2008, Circulation research.

[15]  J. Balligand,et al.  Role of Caveolar Compartmentation in Endothelium-Derived Hyperpolarizing Factor–Mediated Relaxation: Ca2+ Signals and Gap Junction Function Are Regulated by Caveolin in Endothelial Cells , 2008, Circulation.

[16]  T. Takano,et al.  A Central Role of Connexin 43 in Hypoxic Preconditioning , 2008, The Journal of Neuroscience.

[17]  D. Laird,et al.  Caveolin-1 and -2 interact with connexin43 and regulate gap junctional intercellular communication in keratinocytes. , 2007, Molecular biology of the cell.

[18]  B. Uretsky,et al.  Pretreatment with statins may reduce cardiovascular morbidity and mortality after elective surgery and percutaneous coronary intervention: clinical evidence and possible underlying mechanisms. , 2007, American heart journal.

[19]  W. Sessa,et al.  Genetic Evidence Supporting Caveolae Microdomain Regulation of Calcium Entry in Endothelial Cells* , 2007, Journal of Biological Chemistry.

[20]  G. Heusch,et al.  Connexin 43 in ischemic pre- and postconditioning , 2007, Heart Failure Reviews.

[21]  R. Kloner,et al.  Preconditioning in humans , 2007, Heart Failure Reviews.

[22]  I. Grgic,et al.  Evidence for a Functional Role of Endothelial Transient Receptor Potential V4 in Shear Stress–Induced Vasodilatation , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[23]  IvicaGrgic,et al.  Evidence for a Functional Role of Endothelial Transient Receptor Potential V4 in Shear Stress–Induced Vasodilatation , 2006 .

[24]  R. Busse,et al.  Modulation of the Ca2 Permeable Cation Channel TRPV4 by Cytochrome P450 Epoxygenases in Vascular Endothelium , 2005, Circulation research.

[25]  J. López-Barneo,et al.  Decreased Expression of Maxi-K+ Channel β1-Subunit and Altered Vasoregulation in Hypoxia , 2005, Circulation.

[26]  T. Griffith.  Endothelium‐dependent smooth muscle hyperpolarization: do gap junctions provide a unifying hypothesis? , 2004, British journal of pharmacology.

[27]  V. Richard,et al.  Coronary endothelial cells: a target of ischemia reperfusion and its treatment? , 2004, Archives des maladies du coeur et des vaisseaux.

[28]  S. Zahler,et al.  Nitric oxide enhances de novo formation of endothelial gap junctions. , 2003, Cardiovascular research.

[29]  Richard B Schuessler,et al.  Mechanisms of Delayed Electrical Uncoupling Induced by Ischemic Preconditioning , 2003, Circulation research.

[30]  A. Pastuszyn,et al.  Cytochrome p-450 epoxygenase products contribute to attenuated vasoconstriction after chronic hypoxia. , 2003, American journal of physiology. Heart and circulatory physiology.

[31]  P. Kameritsch,et al.  Opposing Effects of Nitric Oxide on Different Connexins Expressed in the Vascular System , 2003, Cell communication & adhesion.

[32]  S. P. Marrelli,et al.  Altered Endothelial Ca2+ Regulation After Ischemia/Reperfusion Produces Potentiated Endothelium-Derived Hyperpolarizing Factor–Mediated Dilations , 2002, Stroke.

[33]  V. Richard,et al.  Endothelial protective effects of preconditioning. , 2002, Cardiovascular research.

[34]  R. Busse,et al.  EDHF: bringing the concepts together. , 2002, Trends in pharmacological sciences.

[35]  Qin Yang,et al.  Hypoxic preconditioning in coronary microarteries: role of EDHF and K+ channel openers. , 2002, The Annals of thoracic surgery.

[36]  A. Ahluwalia,et al.  Protease-Activated Receptor-2 Activation Causes EDHF-Like Coronary Vasodilation: Selective Preservation in Ischemia/Reperfusion Injury: Involvement of Lipoxygenase Products, VR1 Receptors, and C-Fibers , 2002, Circulation research.

[37]  S. Marrelli.  Mechanisms of endothelial P2Y(1)- and P2Y(2)-mediated vasodilatation involve differential [Ca2+]i responses. , 2001, American journal of physiology. Heart and circulatory physiology.

[38]  J. Balligand,et al.  Hsp90 Ensures the Transition from the Early Ca2+-dependent to the Late Phosphorylation-dependent Activation of the Endothelial Nitric-oxide Synthase in Vascular Endothelial Growth Factor-exposed Endothelial Cells* , 2001, The Journal of Biological Chemistry.

[39]  M. Biel,et al.  Lack of an endothelial store-operated Ca2+ current impairs agonist-dependent vasorelaxation in TRP4−/− mice , 2001, Nature Cell Biology.

[40]  D. Yellon,et al.  Ischaemic preconditioning of the vasculature: an overlooked phenomenon for protecting the heart? , 2000, Trends in pharmacological sciences.

[41]  A. Yim,et al.  ATP-sensitive potassium channel openers may mimic the effects of hypoxic preconditioning on the coronary artery. , 2000, The Annals of thoracic surgery.

[42]  J. Czyż,et al.  Gap-junctional coupling measured by flow cytometry. , 2000, Experimental cell research.

[43]  J. Parker,et al.  Nitrate therapy for stable angina pectoris. , 1999, The New England journal of medicine.

[44]  O. Feron,et al.  Modulation of the Endothelial Nitric-oxide Synthase-Caveolin Interaction in Cardiac Myocytes , 1998, The Journal of Biological Chemistry.

[45]  R. Bolli,et al.  Evidence that late preconditioning against myocardial stunning in conscious rabbits is triggered by the generation of nitric oxide. , 1997, Circulation research.

[46]  R. Bolli,et al.  Evidence for an essential role of reactive oxygen species in the genesis of late preconditioning against myocardial stunning in conscious pigs. , 1996, The Journal of clinical investigation.

[47]  A. M. Lefer,et al.  Time course of endothelial dysfunction and myocardial injury during myocardial ischemia and reperfusion in the cat. , 1990, Circulation.

[48]  R. Jennings,et al.  Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. , 1986, Circulation.

[49]  S. Marrelli.  Mechanisms of endothelial P 2 Y 1-and P 2 Y 2-mediated vasodilatation involve differential [ Ca 2 1 ] i responses , 2001 .