Inhibition of T-type Ca2+ Channels by Hydrogen Sulfide

Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 860)


T-type Ca2+ channels are a distinct family of low voltage-activated Ca2+ channels which serve many roles in different tissues. Several studies have implicated them, for example, in the adaptive responses to chronic hypoxia in the cardiovascular and endocrine systems. Hydrogen sulfide (H2S) was more recently discovered as an important signalling molecule involved in many functions, including O2 sensing. Since ion channels are emerging as an important family of target proteins for modulation by H2S, and both T-type Ca2+ channels and H2S are involved in cellular responses to hypoxia, we have investigated whether recombinant and native T-type Ca2+ channels are a target for modulation by H2S. Using patch-clamp electrophysiology, we demonstrate that the H2S donor, NaHS, selectively inhibits Cav3.2 T-type Ca2+ channels heterologously expressed in HEK293 cells, whilst Cav3.1 and Cav3.3 channels were unaffected. Sensitivity of Cav3.2 channels to H2S required the presence of the redox-sensitive extracellular residue H191, which is also required for tonic binding of Zn2+ to this channel. Chelation of Zn2+ using TPEN prevented channel inhibition by H2S. H2S also selectively inhibited native T-type channels (primarily Cav3.2) in sensory dorsal root ganglion neurons. Our data demonstrate a novel target for H2S regulation, the T-type Ca2+ channel Cav3.2. Results have important implications for the proposed pro-nociceptive effects of this gasotransmitter. Implications for the control of cellular responses to hypoxia await further study.


Hydrogen sulfide T-type Ca2+ channel HEK293 cells Sensory neuron Zinc Patch clamp 



This work was supported by The British Heart Foundation.


  1. Boycott HE, Dallas ML, Elies J, Pettinger L, Boyle JP, Scragg JL, Gamper N, Peers C (2013) Carbon monoxide inhibition of Cav3.2 T-type Ca2+ channels reveals tonic modulation by thioredoxin. FASEB J 27:3395–3407PubMedCrossRefGoogle Scholar
  2. Calvert JW, Jha S, Gundewar S, Elrod JW, Ramachandran A, Pattillo CB, Kevil CG, Lefer DJ (2009) Hydrogen sulfide mediates cardioprotection through Nrf2 signaling. Circ Res 105:365–374PubMedCrossRefPubMedCentralGoogle Scholar
  3. Chevalier M, Gilbert G, Roux E, Lory P, Marthan R, Savineau JP, Quignard JF (2014) T-type calcium channels are involved in hypoxic pulmonary hypertension. Cardiovasc Res 103:597–606PubMedCrossRefGoogle Scholar
  4. Fearon IM, Randall AD, Perez-Reyes E, Peers C (2000) Modulation of recombinant T-type Ca2+ channels by hypoxia and glutathione. Pflugers Arch 441:181–188PubMedCrossRefGoogle Scholar
  5. Gong QH, Shi XR, Hong ZY, Pan LL, Liu XH, Zhu YZ (2011) A new hope for neurodegeneration: possible role of hydrogen sulfide. J Alzheimers Dis 24(Suppl 2):173–182PubMedGoogle Scholar
  6. Jevtovic-Todorovic V, Todorovic SM (2006) The role of peripheral T-type calcium channels in pain transmission. Cell Calcium 40:197–203PubMedCrossRefGoogle Scholar
  7. Kawabata A, Ishiki T, Nagasawa K, Yoshida S, Maeda Y, Takahashi T, Sekiguchi F, Wada T, Ichida S, Nishikawa H (2007) Hydrogen sulfide as a novel nociceptive messenger. Pain 132:74–81PubMedCrossRefGoogle Scholar
  8. Kimura Y, Kimura H (2004) Hydrogen sulfide protects neurons from oxidative stress. FASEB J 18:1165–1167PubMedGoogle Scholar
  9. Kimura H, Shibuya N, Kimura Y (2012) Hydrogen sulfide is a signaling molecule and a cytoprotectant. Antioxid Redox Signal 17:45–57PubMedCrossRefPubMedCentralGoogle Scholar
  10. Kirton HM, Pettinger L, Gamper N (2013) Transient overexpression of genes in neurons using nucleofection. Methods Mol Biol 998:55–64PubMedCrossRefGoogle Scholar
  11. Kuga T, Kobayashi S, Hirakawa Y, Kanaide H, Takeshita A (1996) Cell cycle–dependent expression of L- and T-type Ca2+ currents in rat aortic smooth muscle cells in primary culture. Circ Res 79:14–19PubMedCrossRefGoogle Scholar
  12. Levitsky KL, Lopez-Barneo J (2009) Developmental change of T-type Ca2+ channel expression and its role in rat chromaffin cell responsiveness to acute hypoxia. J Physiol 587:1917–1929PubMedCrossRefPubMedCentralGoogle Scholar
  13. Li Q, Sun B, Wang X, Jin Z, Zhou Y, Dong L, Jiang LH, Rong W (2010) A crucial role for hydrogen sulfide in oxygen sensing via modulating large conductance calcium-activated potassium channels. Antioxid Redox Signal 12:1179–1189PubMedCrossRefGoogle Scholar
  14. Li L, Rose P, Moore PK (2011) Hydrogen sulfide and cell signaling. Annu Rev Pharmacol Toxicol 51:169–187PubMedCrossRefGoogle Scholar
  15. Maeda Y, Aoki Y, Sekiguchi F, Matsunami M, Takahashi T, Nishikawa H, Kawabata A (2009) Hyperalgesia induced by spinal and peripheral hydrogen sulfide: evidence for involvement of Cav3.2 T-type calcium channels. Pain 142:127–132PubMedCrossRefGoogle Scholar
  16. Makarenko VV, Nanduri J, Raghuraman G, Fox AP, Gadalla MM, Kumar GK, Snyder SH, Prabhakar NR (2012) Endogenous H2S is required for hypoxic sensing by carotid body glomus cells. Am J Physiol Cell Physiol 303:C916–C923PubMedCrossRefPubMedCentralGoogle Scholar
  17. Matsunami M, Kirishi S, Okui T, Kawabata A (2011) Chelating luminal zinc mimics hydrogen sulfide-evoked colonic pain in mice: possible involvement of T-type calcium channels. Neuroscience 181:257–264PubMedCrossRefGoogle Scholar
  18. Nelson MT, Joksovic PM, Perez-Reyes E, Todorovic SM (2005) The endogenous redox agent L-cysteine induces T-type Ca2+ channel-dependent sensitization of a novel subpopulation of rat peripheral nociceptors. J Neurosci 25:8766–8775PubMedCrossRefGoogle Scholar
  19. Nelson MT, Todorovic SM, Perez-Reyes E (2006) The role of T-type calcium channels in epilepsy and pain. Curr Pharm Des 12:2189–2197PubMedCrossRefGoogle Scholar
  20. Nelson MT, Joksovic PM, Su P, Kang HW, Van DA, Baumgart JP, David LS, Snutch TP, Barrett PQ, Lee JH, Zorumski CF, Perez-Reyes E, Todorovic SM (2007a) Molecular mechanisms of subtype-specific inhibition of neuronal T-type calcium channels by ascorbate. J Neurosci 27:12577–12583PubMedCrossRefGoogle Scholar
  21. Nelson MT, Woo J, Kang HW, Vitko I, Barrett PQ, Perez-Reyes E, Lee JH, Shin HS, Todorovic SM (2007b) Reducing agents sensitize C-type nociceptors by relieving high-affinity zinc inhibition of T-type calcium channels. J Neurosci 27:8250–8260PubMedCrossRefGoogle Scholar
  22. Papapetropoulos A, Whiteman M, Giuseppe C (2015) Pharmacological tools for hydrogen sulfide research: a brief, introductory guide for beginners. Br J Pharmacol 172(6):1633–1637Google Scholar
  23. Paul BD, Snyder SH (2012) H(2)S signalling through protein sulfhydration and beyond. Nat Rev Mol Cell Biol 13:499–507PubMedCrossRefGoogle Scholar
  24. Peers C, Bauer CC, Boyle JP, Scragg JL, Dallas ML (2012) Modulation of ion channels by hydrogen sulfide. Antioxid Redox Signal 17:95–105PubMedCrossRefGoogle Scholar
  25. Peng YJ, Nanduri J, Raghuraman G, Souvannakitti D, Gadalla MM, Kumar GK, Snyder SH, Prabhakar NR (2010) H(2)S mediates O(2) sensing in the carotid body. Proc Natl Acad Sci U S A 107:10719–10724PubMedCrossRefPubMedCentralGoogle Scholar
  26. Perez-Reyes E (2003) Molecular physiology of low-voltage-activated t-type calcium channels. Physiol Rev 83:117–161PubMedCrossRefGoogle Scholar
  27. Peter EA, Shen X, Shah SH, Pardue S, Glawe JD, Zhang WW, Reddy P, Akkus NI, Varma J, Kevil CG (2013) Plasma free H2S levels are elevated in patients with cardiovascular disease. J Am Heart Assoc 2:e000387PubMedCrossRefPubMedCentralGoogle Scholar
  28. Predmore BL, Lefer DJ (2011) Hydrogen sulfide-mediated myocardial pre- and post-conditioning. Expert Rev Clin Pharmacol 4:83–96PubMedCrossRefPubMedCentralGoogle Scholar
  29. Rodman DM, Reese K, Harral J, Fouty B, Wu S, West J, Hoedt-Miller M, Tada Y, Li KX, Cool C, Fagan K, Cribbs L (2005) Low-voltage-activated (T-type) calcium channels control proliferation of human pulmonary artery myocytes. Circ Res 96:864–872PubMedCrossRefGoogle Scholar
  30. Rola R, Szulczyk PJ, Witkowski G (2003) Voltage-dependent Ca2+ currents in rat cardiac dorsal root ganglion neurons. Brain Res 961:171–178PubMedCrossRefGoogle Scholar
  31. Santoni G, Santoni M, Nabissi M (2012) Functional role of T-type calcium channels in tumour growth and progression: prospective in cancer therapy. Br J Pharmacol 166:1244–1246PubMedCrossRefPubMedCentralGoogle Scholar
  32. Shibuya N, Mikami Y, Kimura Y, Nagahara N, Kimura H (2009a) Vascular endothelium expresses 3-mercaptopyruvate sulfurtransferase and produces hydrogen sulfide. J Biochem 146:623–626PubMedCrossRefGoogle Scholar
  33. Shibuya N, Tanaka M, Yoshida M, Ogasawara Y, Togawa T, Ishii K, Kimura H (2009b) 3-Mercaptopyruvate sulfurtransferase produces hydrogen sulfide and bound sulfane sulfur in the brain. Antioxid Redox Signal 11:703–714PubMedCrossRefGoogle Scholar
  34. Szabo C (2007) Hydrogen sulphide and its therapeutic potential. Nat Rev Drug Discov 6:917–935PubMedCrossRefGoogle Scholar
  35. Takahashi T, Aoki Y, Okubo K, Maeda Y, Sekiguchi F, Mitani K, Nishikawa H, Kawabata A (2010) Upregulation of Ca(v)3.2 T-type calcium channels targeted by endogenous hydrogen sulfide contributes to maintenance of neuropathic pain. Pain 150:183–191PubMedCrossRefGoogle Scholar
  36. Talley EM, Cribbs LL, Lee JH, Daud A, Perez-Reyes E, Bayliss DA (1999) Differential distribution of three members of a gene family encoding low voltage-activated (T-type) calcium channels. J Neurosci 19:1895–1911PubMedGoogle Scholar
  37. Todorovic SM, Jevtovic-Todorovic V (2011) T-type voltage-gated calcium channels as targets for the development of novel pain therapies. Br J Pharmacol 163:484–495PubMedCrossRefPubMedCentralGoogle Scholar
  38. Todorovic SM, Jevtovic-Todorovic V, Meyenburg A, Mennerick S, Perez-Reyes E, Romano C, Olney JW, Zorumski CF (2001) Redox modulation of T-type calcium channels in rat peripheral nociceptors. Neuron 31:75–85PubMedCrossRefGoogle Scholar
  39. Wan J, Yamamura A, Zimnicka AM, Voiriot G, Smith KA, Tang H, Ayon RJ, Choudhury MS, Ko EA, Wang J, Wang C, Makino A, Yuan JX (2013) Chronic hypoxia selectively enhances L- and T-type voltage-dependent Ca2+ channel activity in pulmonary artery by upregulating Cav1.2 and Cav3.2. Am J Physiol Lung Cell Mol Physiol 305:L154–L164PubMedCrossRefPubMedCentralGoogle Scholar
  40. Wang R (2012) Physiological implications of hydrogen sulfide: a whiff exploration that blossomed. Physiol Rev 92:791–896PubMedCrossRefGoogle Scholar
  41. Yang G, Wu L, Jiang B, Yang W, Qi J, Cao K, Meng Q, Mustafa AK, Mu W, Zhang S, Snyder SH, Wang R (2008) H2S as a physiologic vasorelaxant: hypertension in mice with deletion of cystathionine gamma-lyase. Science 322:587–590PubMedCrossRefPubMedCentralGoogle Scholar
  42. Zhao W, Zhang J, Lu Y, Wang R (2001) The vasorelaxant effect of H(2)S as a novel endogenous gaseous K(ATP) channel opener. EMBO J 20:6008–6016PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.Leeds Institute of Cardiovascular and Metabolic Medicine, Faculty of Medicine and HealthUniversity of LeedsLeedsUK
  2. 2.School of PharmacyUniversity of ReadingReadingUK
  3. 3.Department of PharmacologyHebei Medical UniversityShijiazhuangChina
  4. 4.School of Biomedical Sciences, Faculty of Biological SciencesUniversity of LeedsLeedsUK

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