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Deletion of T-type calcium channels Cav3.1 or Cav3.2 attenuates endothelial dysfunction in aging mice

  • Molecular and Cellular Mechanisms of Disease
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Abstract

Impairment of endothelial function with aging is accompanied by reduced nitric oxide (NO) production. T-type Cav3.1 channels augment nitric oxide and co-localize with eNOS. Therefore, the hypothesis was that T-type channels contribute to the endothelial dysfunction of aging. Endothelial function was determined in mesenteric arteries (perfusion) and aortae (isometric contraction) of young and old wild-type (WT), Cav3.1, and Cav3.2 knockout mice. NO production was measured by fluorescence imaging in mesenteric arteries. With age, endothelium-dependent subsequent dilatation (following depolarization with KCl) of mesenteric arteries was diminished in the arteries of WT mice, unchanged in Cav3.2−/− preparations but increased in those of Cav3.1−/− mice. NO synthase inhibition abolished the subsequent dilatation in mesenteric arteries and acetylcholine-induced relaxations in aortae. NO levels were significantly reduced in mesenteric arteries of old compared to young WT mice. In Cav3.1−/− and Cav3.2−/− preparations, NO levels increased significantly with age. Relaxations to acetylcholine were significantly smaller in the aortae of old compared to young WT mice, while such responses were comparable in preparations of young and old Cav3.1−/− and Cav3.2−/− mice. The expression of Cav3.1 was significantly reduced in aortae from aged compared to young WT mice. The level of phosphorylated eNOS was significantly increased in aortae from aged Cav3.1−/− mice. In conclusion, T-type calcium channel-deficient mice develop less age-dependent endothelial dysfunction. Changes in NO levels are involved in this phenomenon in WT and Cav3.1−/− mice. These findings suggest that T-type channels play an important role in age-induced endothelial dysfunction.

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References

  1. Abd El-Rahman RR, Harraz OF, Brett SE, Anfinogenova Y, Mufti RE, Goldman D, Welsh DG (2013) Identification of L- and T-type Ca2+ channels in rat cerebral arteries: role in myogenic tone development. Am J Phys Heart Circ Phys 304:H58–H71. https://doi.org/10.1152/ajpheart.00476.2012

    CAS  Google Scholar 

  2. Alp NJ, Channon KM (2004) Regulation of endothelial nitric oxide synthase by tetrahydrobiopterin in vascular disease. Arterioscler Thromb Vasc Biol 24:413–420. https://doi.org/10.1161/01.ATV.0000110785.96039.f6

    Article  CAS  PubMed  Google Scholar 

  3. Andersen H, Jaff MG, Hogh D, Vanhoutte P, Hansen PB (2011) Adenosine elicits an eNOS-independent reduction in arterial blood pressure in conscious mice that involves adenosine A2A receptors. Acta Physiol 203:197–207. https://doi.org/10.1111/j.1748-1716.2010.02218.x

    Article  CAS  Google Scholar 

  4. Ball CJ, Wilson DP, Turner SP, Saint DA, Beltrame JF (2009) Heterogeneity of L- and T-channels in the vasculature: rationale for the efficacy of combined L- and T-blockade. Hypertension 53:654–660. https://doi.org/10.1161/HYPERTENSIONAHA.108.125831

    Article  CAS  PubMed  Google Scholar 

  5. Bjorling K, Morita H, Olsen MF, Prodan A, Hansen PB, Lory P, Holstein-Rathlou NH, Jensen LJ (2013) Myogenic tone is impaired at low arterial pressure in mice deficient in the low-voltage-activated CaV 3.1 T-type Ca(2+) channel. Acta Physiologica (Oxford, England) 207:709–720. https://doi.org/10.1111/apha.12066

    Article  CAS  Google Scholar 

  6. Boittin FX, Gribi F, Serir K, Beny JL (2008) Ca2+-independent PLA2 controls endothelial store-operated Ca2+ entry and vascular tone in intact aorta. Am J Phys Heart Circ Phys 295:H2466–H2474. https://doi.org/10.1152/ajpheart.00639.2008

    CAS  Google Scholar 

  7. Boulanger CM, Desta B, Clozel JP, Vanhoutte PM (1994) Chronic treatment with the CA2+ channel inhibitor RO 40-5967 potentiates endothelium-dependent relaxations in the aorta of the hypertensive salt sensitive Dahl rat. Blood Press 3:193–196

    Article  CAS  PubMed  Google Scholar 

  8. Boulanger CM, Nakashima M, Olmos L, Joly G, Vanhoutte PM (1994) Effects of the Ca2+ antagonist RO 40-5967 on endothelium-dependent responses of isolated arteries. J Cardiovasc Pharmacol 23:869–876

    Article  CAS  PubMed  Google Scholar 

  9. Braunstein TH, Inoue R, Cribbs L, Oike M, Ito Y, Holstein-Rathlou NH, Jensen LJ (2009) The role of L- and T-type calcium channels in local and remote calcium responses in rat mesenteric terminal arterioles. J Vasc Res 46:138–151. https://doi.org/10.1159/000151767

    Article  CAS  PubMed  Google Scholar 

  10. Cai H, Harrison DG (2000) Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res 87:840–844

    Article  CAS  PubMed  Google Scholar 

  11. Cazade M, Bidaud I, Hansen PB, Lory P, Chemin J (2014) 5,6-EET potently inhibits T-type calcium channels: implication in the regulation of the vascular tone. Pflugers Archiv : Eur J Physiol 466:1759–1768. https://doi.org/10.1007/s00424-013-1411-0

    Article  CAS  Google Scholar 

  12. Chen CC, Lamping KG, Nuno DW, Barresi R, Prouty SJ, Lavoie JL, Cribbs LL, England SK, Sigmund CD, Weiss RM, Williamson RA, Hill JA, Campbell KP (2003) Abnormal coronary function in mice deficient in alpha1H T-type Ca2+ channels. Science (New York, NY) 302:1416–1418. https://doi.org/10.1126/science.1089268

    Article  CAS  Google Scholar 

  13. Davidge ST, Ojimba J, McLaughlin MK (1998) Vascular function in the vitamin E-deprived rat: an interaction between nitric oxide and superoxide anions. Hypertension 31:830–835

    Article  CAS  PubMed  Google Scholar 

  14. Deacon K, Knox AJ (2010) Endothelin-1 (ET-1) increases the expression of remodeling genes in vascular smooth muscle through linked calcium and cAMP pathways: role of a phospholipase A(2)(cPLA(2))/cyclooxygenase-2 (COX-2)/prostacyclin receptor-dependent autocrine loop. J Biol Chem 285:25913–25927. https://doi.org/10.1074/jbc.M110.139485

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Feletou M, Kohler R, Vanhoutte PM (2012) Nitric oxide: orchestrator of endothelium-dependent responses. Ann Med 44:694–716. https://doi.org/10.3109/07853890.2011.585658

    Article  CAS  PubMed  Google Scholar 

  16. Fujii K, Ohmori S, Tominaga M, Abe I, Takata Y, Ohya Y, Kobayashi K, Fujishima M (1993) Age-related changes in endothelium-dependent hyperpolarization in the rat mesenteric artery. Am J Phys 265:H509–H516

    CAS  Google Scholar 

  17. Furchgott RF, Vanhoutte PM (1989) Endothelium-derived relaxing and contracting factors. FASEB J : Off Publ Fed Am Soc Exp Biol 3:2007–2018

    Article  CAS  Google Scholar 

  18. Hansen PB, Jensen BL, Andreasen D, Skott O (2001) Differential expression of T- and L-type voltage-dependent calcium channels in renal resistance vessels. Circ Res 89:630–638

    Article  CAS  PubMed  Google Scholar 

  19. Hansen PB, Poulsen CB, Walter S, Marcussen N, Cribbs LL, Skott O, Jensen BL (2011) Functional importance of L- and P/Q-type voltage-gated calcium channels in human renal vasculature. Hypertension 58:464–470. https://doi.org/10.1161/hypertensionaha.111.170845

    Article  CAS  PubMed  Google Scholar 

  20. Harraz OF, Abd El-Rahman RR, Bigdely-Shamloo K, Wilson SM, Brett SE, Romero M, Gonzales AL, Earley S, Vigmond EJ, Nygren A, Menon BK, Mufti RE, Watson T, Starreveld Y, Furstenhaupt T, Muellerleile PR, Kurjiaka DT, Kyle BD, Braun AP, Welsh DG (2014) Ca(V)3.2 channels and the induction of negative feedback in cerebral arteries. Circ Res 115:650–661. https://doi.org/10.1161/circresaha.114.304056

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Howitt L, Kuo IY, Ellis A, Chaston DJ, Shin HS, Hansen PB, Hill CE (2013) Chronic deficit in nitric oxide elicits oxidative stress and augments T-type calcium-channel contribution to vascular tone of rodent arteries and arterioles. Cardiovasc Res 98:449–457. https://doi.org/10.1093/cvr/cvt043

    Article  CAS  PubMed  Google Scholar 

  22. Howitt L, Matthaei KI, Drummond GR, Hill CE (2014) Nox1 upregulates the function of vascular T-type calcium channels following chronic nitric oxide deficit. Pflugers Archiv : Eur J Physiol. https://doi.org/10.1007/s00424-014-1548-5

  23. Imaoka Y, Osanai T, Kamada T, Mio Y, Satoh K, Okumura K (1999) Nitric oxide-dependent vasodilator mechanism is not impaired by hypertension but is diminished with aging in the rat aorta. J Cardiovasc Pharmacol 33:756–761

    Article  CAS  PubMed  Google Scholar 

  24. Kluge MA, Fetterman JL, Vita JA (2013) Mitochondria and endothelial function. Circ Res 112:1171–1188. https://doi.org/10.1161/CIRCRESAHA.111.300233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Koga T, Takata Y, Kobayashi K, Takishita S, Yamashita Y, Fujishima M (1988) Ageing suppresses endothelium-dependent relaxation and generates contraction mediated by the muscarinic receptors in vascular smooth muscle of normotensive Wistar-Kyoto and spontaneously hypertensive rats. J Hypertens Suppl : Off J Int Soc Hypertens 6:S243–S245

    Article  CAS  Google Scholar 

  26. Koh KK, Quon MJ, Lee SJ, Han SH, Ahn JY, Kim JA, Chung WJ, Lee Y, Shin EK (2007) Efonidipine simultaneously improves blood pressure, endothelial function, and metabolic parameters in nondiabetic patients with hypertension. Diabetes Care 30:1605–1607. https://doi.org/10.2337/dc06-2267

    Article  CAS  PubMed  Google Scholar 

  27. Kung CF, Moreau P, Takase H, Luscher TF (1995) L-NAME hypertension alters endothelial and smooth muscle function in rat aorta. Prevention by trandolapril and verapamil. Hypertens 26:744–751

    Article  CAS  Google Scholar 

  28. Lee J, Kim D, Shin HS (2004) Lack of delta waves and sleep disturbances during non-rapid eye movement sleep in mice lacking alpha1G-subunit of T-type calcium channels. Proc Natl Acad Sci U S A 101:18195–18199. https://doi.org/10.1073/pnas.0408089101

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Liu S, Premont RT, Rockey DC (2014) Endothelial nitric-oxide synthase (eNOS) is activated through G-protein-coupled receptor kinase-interacting protein 1 (GIT1) tyrosine phosphorylation and Src protein. J Biol Chem 289:18163–18174. https://doi.org/10.1074/jbc.M113.521203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Luscher TF, Vanhoutte PM (1986) Endothelium-dependent contractions to acetylcholine in the aorta of the spontaneously hypertensive rat. Hypertens 8:344–348

    Article  CAS  Google Scholar 

  31. Matz RL, Schott C, Stoclet JC, Andriantsitohaina R (2000) Age-related endothelial dysfunction with respect to nitric oxide, endothelium-derived hyperpolarizing factor and cyclooxygenase products. Physiol Res / Academia Scientiarum Bohemoslovaca 49:11–18

    CAS  Google Scholar 

  32. Miller VM, Vanhoutte PM (1985) Endothelium-dependent contractions to arachidonic acid are mediated by products of cyclooxygenase. Am J Phys 248:H432–H437

    CAS  Google Scholar 

  33. Mombouli JV, Vanhoutte PM (1993) Purinergic endothelium-dependent and -independent contractions in rat aorta. Hypertension 22:577–583

    Article  CAS  PubMed  Google Scholar 

  34. Oshima T, Ozono R, Yano Y, Higashi Y, Teragawa H, Miho N, Ishida T, Ishida M, Yoshizumi M, Kambe M (2005) Beneficial effect of T-type calcium channel blockers on endothelial function in patients with essential hypertension. Hypertens Res : Off J Jpn Soc Hypertens 28:889–894. https://doi.org/10.1291/hypres.28.889

    Article  CAS  Google Scholar 

  35. Pearson PJ, Vanhoutte PM (1993) Vasodilator and vasoconstrictor substances produced by the endothelium. Rev Physiol Biochem Pharmacol 122:1–67

    Article  CAS  PubMed  Google Scholar 

  36. Poulsen CB, Al-Mashhadi RH, Cribbs LL, Skott O, Hansen PB (2011) T-type voltage-gated calcium channels regulate the tone of mouse efferent arterioles. Kidney Int 79:443–451. https://doi.org/10.1038/ki.2010.429

    Article  CAS  PubMed  Google Scholar 

  37. Rapoport RM, Williams SP (1996) Role of prostaglandins in acetylcholine-induced contraction of aorta from spontaneously hypertensive and Wistar-Kyoto rats. Hypertens 28:64–75

    Article  CAS  Google Scholar 

  38. Rubanyi GM, Vanhoutte PM (1988) Potassium-induced release of endothelium-derived relaxing factor from canine femoral arteries. Circ Res 62:1098–1103

    Article  CAS  PubMed  Google Scholar 

  39. Santhanam L, Christianson DW, Nyhan D, Berkowitz DE (2008) Arginase and vascular aging. J Appl Physiol (Bethesda, Md : 1985) 105:1632–1642. https://doi.org/10.1152/japplphysiol.90627.2008

    Article  CAS  Google Scholar 

  40. Schjerning J, Uhrenholt TR, Svenningsen P, Vanhoutte PM, Skott O, Jensen BL, Hansen PB (2013) Histamine-dependent prolongation by aldosterone of vasoconstriction in isolated small mesenteric arteries of the mouse. Am J Phys Heart Circ Phys 304:H1094–H1102. https://doi.org/10.1152/ajpheart.00524.2012

    CAS  Google Scholar 

  41. Seegers HC, Gross RW, Boyle WA (2002) Calcium-independent phospholipase A(2)-derived arachidonic acid is essential for endothelium-dependent relaxation by acetylcholine. J Pharmacol Exp Ther 302:918–923

    Article  CAS  PubMed  Google Scholar 

  42. Shi Y, Feletou M, Ku DD, Man RY, Vanhoutte PM (2007) The calcium ionophore A23187 induces endothelium-dependent contractions in femoral arteries from rats with streptozotocin-induced diabetes. Br J Pharmacol 150:624–632. https://doi.org/10.1038/sj.bjp.0706999

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Shi Y, Man RY, Vanhoutte PM (2008) Two isoforms of cyclooxygenase contribute to augmented endothelium-dependent contractions in femoral arteries of 1-year-old rats. Acta Pharmacol Sin 29:185–192. https://doi.org/10.1111/j.1745-7254.2008.00749.x

    Article  CAS  PubMed  Google Scholar 

  44. Shi Y, So KF, Man RY, Vanhoutte PM (2007) Oxygen-derived free radicals mediate endothelium-dependent contractions in femoral arteries of rats with streptozotocin-induced diabetes. Br J Pharmacol 152:1033–1041. https://doi.org/10.1038/sj.bjp.0707439

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Shimizu I, Toda N (1986) Alterations with age of the response to vasodilator agents in isolated mesenteric arteries of the beagle. Br J Pharmacol 89:769–778

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Sobey CG (2001) Potassium channel function in vascular disease. Arterioscler Thromb Vasc Biol 21:28–38

    Article  CAS  PubMed  Google Scholar 

  47. Svenningsen P, Andersen K, Thuesen AD, Shin HS, Vanhoutte PM, Skott O, Jensen BL, Hill C, Hansen PB (2014) T-type Ca(2+) channels facilitate NO-formation, vasodilatation and NO-mediated modulation of blood pressure. Pflugers Archiv : Eur J Physiol 466:2205–2214. https://doi.org/10.1007/s00424-014-1492-4

    Article  CAS  Google Scholar 

  48. Tang EH, Leung FP, Huang Y, Feletou M, So KF, Man RY, Vanhoutte PM (2007) Calcium and reactive oxygen species increase in endothelial cells in response to releasers of endothelium-derived contracting factor. Br J Pharmacol 151:15–23. https://doi.org/10.1038/sj.bjp.0707190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Thors B, Halldorsson H, Thorgeirsson G (2004) Thrombin and histamine stimulate endothelial nitric-oxide synthase phosphorylation at Ser1177 via an AMPK mediated pathway independent of PI3K-Akt. FEBS Lett 573:175–180. https://doi.org/10.1016/j.febslet.2004.07.078

    Article  CAS  PubMed  Google Scholar 

  50. van der Loo B, Labugger R, Skepper JN, Bachschmid M, Kilo J, Powell JM, Palacios-Callender M, Erusalimsky JD, Quaschning T, Malinski T, Gygi D, Ullrich V, Luscher TF (2000) Enhanced peroxynitrite formation is associated with vascular aging. J Exp Med 192:1731–1744

    Article  PubMed  PubMed Central  Google Scholar 

  51. Vanhoutte PM (2013) One or two, does it matter as long as the arterial wall is coxygenated? Hypertens 62:244–246. https://doi.org/10.1161/hypertensionaha.113.01566

    Article  CAS  Google Scholar 

  52. Wang L, Yin J, Nickles HT, Ranke H, Tabuchi A, Hoffmann J, Tabeling C, Barbosa-Sicard E, Chanson M, Kwak BR, Shin HS, Wu S, Isakson BE, Witzenrath M, de Wit C, Fleming I, Kuppe H, Kuebler WM (2012) Hypoxic pulmonary vasoconstriction requires connexin 40-mediated endothelial signal conduction. J Clin Invest 122:4218–4230. https://doi.org/10.1172/jci59176

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Widlansky ME, Gutterman DD (2011) Regulation of endothelial function by mitochondrial reactive oxygen species. Antioxid Redox Signal 15:1517–1530. https://doi.org/10.1089/ars.2010.3642

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Wong MS, Man RY, Vanhoutte PM (2010) Calcium-independent phospholipase A(2) plays a key role in the endothelium-dependent contractions to acetylcholine in the aorta of the spontaneously hypertensive rat. Am J Phys Heart Circ Phys 298:H1260–H1266. https://doi.org/10.1152/ajpheart.01068.2009

    CAS  Google Scholar 

  55. Yang D, Feletou M, Boulanger CM, Wu HF, Levens N, Zhang JN, Vanhoutte PM (2002) Oxygen-derived free radicals mediate endothelium-dependent contractions to acetylcholine in aortas from spontaneously hypertensive rats. Br J Pharmacol 136:104–110. https://doi.org/10.1038/sj.bjp.0704669

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Zhou E, Qing D, Li J (2010) Age-associated endothelial dysfunction in rat mesenteric arteries: roles of calcium-activated K(+) channels (K(ca)). Physiol Res / Academia Scientiarum Bohemoslovaca 59:499–508

    CAS  Google Scholar 

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Acknowledgements

We would like to thank Vivi Monrad and Mie Rytz for expert technical assistance. Also, we thank Hee-Sup Shin, Center for Cognition and Sociality, Korea, for the Cav3.1−/− mice and Philippe Lory, University de Montpellier, France.

Funding

This work was supported by grants from the Danish Medical Research Council (11-107552), The Danish Heart Foundation (11-04-R84-A3492-22663), and Novo Nordic Foundation (13273).

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Authors and Affiliations

  1. Cardiovascular and Renal Research, University of Southern Denmark, J. B. Winsløwsvej 21, 3., 5000, Odense C, Denmark

    Anne D. Thuesen, Kenneth Andersen, Kristina S. Lyngsø & Pernille B. L. Hansen

  2. State Key Laboratory for Biopharmaceutical Technology and Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China

    Paul M. Vanhoutte

  3. Department of Clinical Genetic, Odense University Hospital, 5000, Odense, Denmark

    Mark Burton & Charlotte Brasch-Andersen

  4. Cardiovascular and Metabolic Disease Innovative Medicines, AstraZeneca, Gothenburg, Sweden

    Pernille B. L. Hansen

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  1. Anne D. Thuesen
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Thuesen, A.D., Andersen, K., Lyngsø, K.S. et al. Deletion of T-type calcium channels Cav3.1 or Cav3.2 attenuates endothelial dysfunction in aging mice. Pflugers Arch - Eur J Physiol 470, 355–365 (2018). https://doi.org/10.1007/s00424-017-2068-x

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