Advertisement

Anoctamins support calcium-dependent chloride secretion by facilitating calcium signaling in adult mouse intestine

  • Rainer Schreiber
  • Diana Faria
  • Boris V. Skryabin
  • Podchanart Wanitchakool
  • Jason R. Rock
  • Karl KunzelmannEmail author
Ion channels, receptors and transporters

Abstract

Intestinal epithelial electrolyte secretion is activated by increase in intracellular cAMP or Ca2+ and opening of apical Cl channels. In infants and young animals, but not in adults, Ca2+-activated chloride channels may cause secretory diarrhea during rotavirus infection. While detailed knowledge exists concerning the contribution of cAMP-activated cystic fibrosis transmembrane conductance regulator (CFTR) channels, analysis of the role of Ca2+-dependent Cl channels became possible through identification of the anoctamin (TMEM16) family of proteins. We demonstrate expression of several anoctamin paralogues in mouse small and large intestines. Using intestinal-specific mouse knockout models for anoctamin 1 (Ano1) and anoctamin 10 (Ano10) and a conventional knockout model for anoctamin 6 (Ano6), we demonstrate the role of anoctamins for Ca2+-dependent Cl secretion induced by the muscarinic agonist carbachol (CCH). Ano1 is preferentially expressed in the ileum and large intestine, where it supports Ca2+-activated Cl secretion. In contrast, Ano10 is essential for Ca2+-dependent Cl secretion in jejunum, where expression of Ano1 was not detected. Although broadly expressed, Ano6 has no role in intestinal cholinergic Cl secretion. Ano1 is located in a basolateral compartment/membrane rather than in the apical membrane, where it supports CCH-induced Ca2+ increase, while the essential and possibly only apical Cl channel is CFTR. These results define a new role of Ano1 for intestinal Ca2+-dependent Cl secretion and demonstrate for the first time a contribution of Ano10 to intestinal transport.

Keywords

TMEM16A TMEM16F TMEM16K Anoctamin 1 Anoctamin 6 Anoctamin 10 Ano1 Ano6 Ano10 Ca2+-activated Cl channels Colon Small intestine Ileum Jejunum chloride secretion 

Notes

Acknowledgments

This study was supported by DFG SFB699A7 and Wilhelm-Sander Stiftung Ano6 and Deutsche Krebshilfe Projekt 109438. We gratefully acknowledge the generous supply of the Ano6−/− mice by Prof. Dr. A. Vortkamp (Department Entwicklungsbiologie, University of Essen, Essen, Germany) and Ano-1 antibodies by Prof. Dr. Brian Harfe (University of Florida at Gainesville, Gainesville, USA).

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

424_2014_1559_MOESM1_ESM.docx (16 kb)
Supplementary Table S1 (DOCX 15 kb)
424_2014_1559_MOESM2_ESM.pdf (75 kb)
Supplementary Fig. S1 (PDF 75 kb)
424_2014_1559_MOESM3_ESM.pdf (499 kb)
Supplementary Fig. S2 (PDF 499 kb)
424_2014_1559_MOESM4_ESM.pdf (575 kb)
Supplementary Fig. S3 (PDF 575 kb)
424_2014_1559_MOESM5_ESM.pdf (108 kb)
Supplementary Fig. S4 (PDF 107 kb)
424_2014_1559_MOESM6_ESM.pdf (55 kb)
Supplementary Fig. S5 (PDF 55 kb)

References

  1. 1.
    Almaca J, Tian Y, AlDehni F, Ousingsawat J, Kongsuphol P, Rock JR, Harfe BD, Schreiber R, Kunzelmann K (2009) TMEM16 proteins produce volume regulated chloride currents that are reduced in mice lacking TMEM16A. J Biol Chem 284:28571–28578CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Ball JM, Tian P, Zeng CQ, Morris AP, Estes MK (1996) Age-dependent diarrhea induced by a rotaviral nonstructural glycoprotein. Science 272:101–104CrossRefPubMedGoogle Scholar
  3. 3.
    Barro Soria R, AlDehni F, Almaca J, Witzgall R, Schreiber R, Kunzelmann K (2009) ER localized bestrophin1 acts as a counter-ion channel to activate Ca2+ dependent ion channels TMEM16A and SK4. Pflugers Arch 459:485–497CrossRefPubMedGoogle Scholar
  4. 4.
    Billet A, Hanrahan JW (2013) The secret life of CFTR as a calcium-activated chloride channel. J Physiol 591(21):5273–5278Google Scholar
  5. 5.
    Billig GM, Pál B, Fidzinski P, Jentsch TJ (2011) Ca2+−activated Cl currents are dispensable for olfaction. Nat Neurosci 14:763–769CrossRefPubMedGoogle Scholar
  6. 6.
    Caputo A, Caci E, Ferrera L, Pedemonte N, Barsanti C, Sondo E, Pfeffer U, Ravazzolo R, Zegarra-Moran O, Galietta LJ (2008) TMEM16A, a membrane protein associated with calcium-dependent chloride channel activity. Science 322:590–594CrossRefPubMedGoogle Scholar
  7. 7.
    Chamova T, Florez L, Guergueltcheva V, Raycheva M, Kaneva R, Lochmuller H, Kalaydjieva L, Tournev I (2012) ANO10 c.1150_1151del is a founder mutation causing autosomal recessive cerebellar ataxia in Roma/Gypsies. J Neurol 259:906–911CrossRefPubMedGoogle Scholar
  8. 8.
    Dekkers JF, Wiegerinck CL, De Jonge HR, Bronsveld I, Janssens HM, de Winter-de Groot KM, Brandsma AM, de Jong NW, Bijvelds MJ, Scholte BJ, Nieuwenhuis EE, van den Brink S, Clevers H, van der Ent CK, Middendorp S, Beekman JM (2013) A functional CFTR assay using primary cystic fibrosis intestinal organoids. Nat Med 19:939–945CrossRefPubMedGoogle Scholar
  9. 9.
    Dong Y, Zeng CQ, Ball JM, Estes MK, Morris AP (1997) The rotavirus enterotoxin NSP4 mobilizes intracellular calcium in human intestinal cells by stimulating phospholipase C-mediated inositol 1,4,5- trisphosphate production. Proc Natl Acad Sci U S A 94:3960–3965CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Ehlen HW, Chinenkova M, Moser M, Munter HM, Krause Y, Gross S, Brachvogel B, Wuelling M, Kornak U, Vortkamp A (2012) Inactivation of Anoctamin-6/Tmem16f, a regulator of phosphatidylserine scrambling in osteoblasts, leads to decreased mineral deposition in skeletal tissues. J Bone Miner Res 28:246–259CrossRefGoogle Scholar
  11. 11.
    Elvers M, Stegner D, Hagedorn I, Kleinschnitz C, Braun A, Kuijpers ME, Boesl M, Chen Q, Heemskerk JW, Stoll G, Frohman MA, Nieswandt B (2010) Impaired alpha(IIb)beta(3) integrin activation and shear-dependent thrombus formation in mice lacking phospholipase D1. Sci Signal 3:ra1CrossRefPubMedCentralPubMedGoogle Scholar
  12. 12.
    Faria D, Schlatter E, Witzgall R, Grahammer F, Bandulik S, Schweda F, Bierer S, Rock JR, Heitzmann D, Kunzelmann K, Schreiber R (2013) The calcium activated chloride channel Anoctamin 1 contributes to the regulation of renal function. Kindey Int 85(6):1369–1381Google Scholar
  13. 13.
    Faria D, Schreiber R, Kunzelmann K (2009) CFTR is activated through stimulation of purinergic P2Y2 receptors. Pflugers Arch 457:1373–1380CrossRefPubMedGoogle Scholar
  14. 14.
    Frizzell RA, Hanrahan JW (2012) Physiology of epithelial chloride and fluid secretion. Cold Spring Harb Perspect Med 2:a009563CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    Grubb S, Poulsen KA, Juul CA, Kyed T, Klausen TK, Larsen EH, Hoffmann EK (2013) TMEM16F (Anoctamin 6), an anion channel of delayed Ca2+ activation. J Gen Physiol 141:585–600CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    He Q, Halm ST, Zhang J, Halm DR (2011) Activation of the basolateral membrane Cl conductance essential for electrogenic K secretion suppresses electrogenic Cl secretion. Exp Physiol 96:305–316CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Heemskerk JW, Bevers EM, Lindhout T (2002) Platelet activation and blood coagulation. Thromb Haemost 88:186–193PubMedGoogle Scholar
  18. 18.
    Hennig B, Schultheiss G, Kunzelmann K, Diener M (2008) Ca(2+ )-induced Cl (−) efflux at rat distal colonic epithelium. J Membr Biol 221:61–72CrossRefPubMedGoogle Scholar
  19. 19.
    Hogan DL, Crombie DL, Isenberg JI, Svendsen P, Schaffalitzky de Muckadell OB, Ainsworth MA (1997) CFTR mediates cAMP- and Ca2+−activated duodenal epithelial HCO3- secretion. Am J Physiol 272:G872–G878PubMedGoogle Scholar
  20. 20.
    Juul CA, Grubb S, Poulsen KA, Kyed T, Hashem N, Lambert IH, Larsen EH, Hoffmann EK (2014) Anoctamin 6 differs from VRAC and VSOAC but is involved in apoptosis and supports volume regulation in the presence of Ca. Pflugers Arch [Epub ahead of print]Google Scholar
  21. 21.
    Kmit A, van Kruchten R, Ousingsawat J, Mattheij NJ, Senden-Gijsbers B, Heemskerk JW, Bevers EM, Kunzelmann K (2013) Calcium-activated and apoptotic phospholipid scrambling induced by Ano6 can occur independently of Ano6 ion currents. Cell Death Dis 4:e611CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Ko EA, Jin BJ, Namkung W, Ma T, Thiagarajah JR, and Verkman AS (2013) Chloride channel inhibition by a red wine extract and a synthetic small molecule prevents rotaviral secretory diarrhoea in neonatal mice. Gut 63(7):1120–1129Google Scholar
  23. 23.
    Kunzelmann K, Kongsuphol P, AlDehni F, Tian Y, Ousingsawat J, Warth R, Schreiber R (2009) Bestrophin and TMEM16—Ca2+ activated Cl channels with different functions. Cell Calcium 46:233–241CrossRefPubMedGoogle Scholar
  24. 24.
    Kunzelmann K, Mall M (2002) Electrolyte transport in the colon: mechanisms and implications for disease. Physiol Rev 82:245–289PubMedGoogle Scholar
  25. 25.
    Kunzelmann K, Mehta A (2013) CFTR: a hub for kinases and cross-talk of cAMP and Ca. FEBS J 280:4417–4429CrossRefPubMedGoogle Scholar
  26. 26.
    Kunzelmann K, Nilius B, Owsianik G, Schreiber R, Ousingsawat J, Sirianant L, Wanitchakool P, Bevers EM, Heemskerk JW (2013) Molecular functions of anoctamin 6 (TMEM16F): A chloride channel, cation channel or phospholipid scramblase? Pflügers Arch 466(3):407–14Google Scholar
  27. 27.
    Kunzelmann K, Tian Y, Martins JR, Faria D, Kongsuphol P, Ousingsawat J, Thevenod F, Roussa E, Rock JR, Schreiber R (2011) Anoctamins. Pflugers Arch 462:195–208CrossRefPubMedGoogle Scholar
  28. 28.
    Kunzelmann K, Tian Y, Martins JR, Faria D, Kongsuphol P, Ousingsawat J, Wolf L, Schreiber R (2012) Cells in focus: airway epithelial cells-Functional links between CFTR and anoctamin dependent Cl(−) secretion. Int J Biochem Cell Biol 44:1897–1900CrossRefPubMedGoogle Scholar
  29. 29.
    Mall M, Bleich M, Greger R, Schürlein M, Kühr J, Seydewitz HH, Brandis M, Kunzelmann K (1998) Cholinergic ion secretion in human colon requires co-activation by cAMP. Am J Physiol 275:G1274–G1281PubMedGoogle Scholar
  30. 30.
    Malvezzi M, Chalat M, Janjusevic R, Picollo A, Terashima H, Menon AK, Accardi A (2013) Ca(2+)-dependent phospholipid scrambling by a reconstituted TMEM16 ion channel. Nat Commun 4:2367CrossRefPubMedCentralPubMedGoogle Scholar
  31. 31.
    Martins JR, Faria D, Kongsuphol P, Reisch B, Schreiber R, Kunzelmann K (2011) Anoctamin 6 is an essential component of the outwardly rectifying chloride channel. Proc Natl Acad Sci U S A 108:18168–18172CrossRefPubMedCentralPubMedGoogle Scholar
  32. 32.
    Maruyama H, Morino H, Miyamoto R, Murakami N, Hamano T, Kawakami H (2013) Exome sequencing reveals a novel ANO10 mutation in a Japanese patient with autosomal recessive spinocerebellar ataxia. Clin Genet 85(3):296–7Google Scholar
  33. 33.
    Murek M, Kopic S, Geibel J (2010) Evidence for intestinal chloride secretion. Exp Physiol 95:471–478CrossRefPubMedGoogle Scholar
  34. 34.
    Namkung W, Finkbeiner WE, Verkman AS (2010) CFTR-Adenylyl Cyclase I association is responsible for UTP activation of CFTR in well-differentiated primary human bronchial cell cultures. Mol Biol Cell 21:2639–2648CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Neussert R, Muller C, Milenkovic VM, Strauss O (2010) The presence of bestrophin-1 modulates the Ca(2+) recruitment from Ca (2+) stores in the ER. Pflugers Arch 460:163–175CrossRefPubMedGoogle Scholar
  36. 36.
    Ousingsawat J, Kongsuphol P, Schreiber R, Kunzelmann K (2011) CFTR and TMEM16A are separate but functionally related Cl channels. Cell Physiol Biochem 28:715–724CrossRefPubMedGoogle Scholar
  37. 37.
    Ousingsawat J, Martins JR, Schreiber R, Rock JR, Harfe BD, Kunzelmann K (2009) Loss of TMEM16A causes a defect in epithelial Ca2+ dependent chloride transport. J Biol Chem 284:28698–28703CrossRefPubMedCentralPubMedGoogle Scholar
  38. 38.
    Ousingsawat J, Tian Y, AlDehni F, Roussa E, Schreiber R, Mirza M, Cook DI, Kunzelmann K (2011) Rotavirus toxin NSP4 activates the calcium dependent chloride channel TMEM16A and inhibits absorptive Na+transport. Pflugers Arch 461:579–589CrossRefPubMedGoogle Scholar
  39. 39.
    Pifferi S, Dibattista M, Menini A (2009) TMEM16B induces chloride currents activated by calcium in mammalian cells. Pflugers Arch 458:1023–1038CrossRefPubMedGoogle Scholar
  40. 40.
    Puntheeranurak S, Schreiber R, Spitzner M, Ousingsawat J, Krishnamra N, Kunzelmann K (2007) Control of ion transport in mouse proximal and distal colon by prolactin. Cell Physiol Biochem 19:77–88CrossRefPubMedGoogle Scholar
  41. 41.
    Sailer A, Houlden H (2012) Recent advances in the genetics of cerebellar ataxias. Curr Neurol Neurosci Rep 12:227–236CrossRefPubMedGoogle Scholar
  42. 42.
    Schreiber R, Kunzelmann K (2005) Purinergic P2Y6 receptors induce Ca2+ and CFTR dependent Cl secretion in mouse trachea. Cell Physiol Biochem 16:99–108CrossRefPubMedGoogle Scholar
  43. 43.
    Schreiber R, Uliyakina I, Kongsuphol P, Warth R, Mirza M, Martins JR, Kunzelmann K (2010) Expression and function of epithelial anoctamins. J Biol Chem 285:7838–7845CrossRefPubMedCentralPubMedGoogle Scholar
  44. 44.
    Schroeder BC, Cheng T, Jan YN, Jan LY (2008) Expression cloning of TMEM16A as a calcium-activated chloride channel subunit. Cell 134:1019–1029CrossRefPubMedCentralPubMedGoogle Scholar
  45. 45.
    Seidler U, Blumenstein I, Kretz A, Viellard-Baron D, Rossmann H, Colledge WH, Evans M, Ratcliff R, Gregor M (1997) A functional CFTR protein is required for mouse intestinal cAMP-, cGMP- and Ca(2+)-dependent HCO3- secretion. J Physiol 505:411–423CrossRefPubMedCentralPubMedGoogle Scholar
  46. 46.
    Shimizu T, Lehara T, Sato K, Fujii T, Sakai H, Okada Y (2013) TMEM16F is a component of a Ca2+−activated Cl channel but not a volume-sensitive outwardly rectifying Cl channel. Am J Physiol Cell Physiol 304:C748–C759CrossRefPubMedGoogle Scholar
  47. 47.
    Stohr H, Heisig JB, Benz PM, Schoberl S, Milenkovic VM, Strauss O, Aartsen WM, Wijnholds J, Weber BH, Schulz HL (2009) TMEM16B, a novel protein with calcium-dependent chloride channel activity, associates with a presynaptic protein complex in photoreceptor terminals. J Neurosci 29:6809–6818CrossRefPubMedGoogle Scholar
  48. 48.
    Strauss O, Muller C, Reichhart N, Tamm ER, Gomez NM (2014) The role of bestrophin-1 in intracellular ca(2+) signaling. Adv Exp Med Biol 801:113–119CrossRefPubMedGoogle Scholar
  49. 49.
    Tian Y, Schreiber R, Kunzelmann K (2012) Anoctamins are a family of Ca2+ activated Cl channels. J Cell Sci 125:4991–4998CrossRefPubMedGoogle Scholar
  50. 50.
    Vermeer S, Hoischen A, Meijer RP, Gilissen C, Neveling K, Wieskamp N, de Brouwer A, Koenig M, Anheim M, Assoum M, Drouot N, Todorovic S, Milic-Rasic V, Lochmuller H, Stevanin G, Goizet C, David A, Durr A, Brice A, Kremer B, van de Warrenburg BP, Schijvenaars MM, Heister A, Kwint M, Arts P, van der Wijst J, Veltman J, Kamsteeg EJ, Scheffer H, Knoers N (2010) Targeted next-generation sequencing of a 12.5 Mb homozygous region reveals ANO10 mutations in patients with autosomal-recessive cerebellar ataxia. Am J Hum Genet 87:813–819CrossRefPubMedCentralPubMedGoogle Scholar
  51. 51.
    Viitanen T, Sukumaran P, Lof C, Tornquist K (2012) Functional coupling of TRPC2 cation channels and the calcium-activated anion channels in rat thyroid cells: implications for iodide homeostasis. J Cell Physiol 228(4):814–823Google Scholar
  52. 52.
    Watt SA, Kular G, Fleming IN, Downes CP, Lucocq JM (2002) Subcellular localization of phosphatidylinositol 4,5-bisphosphate using the pleckstrin homology domain of phospholipase C delta1. Biochem J 363(Pt.3):657–666CrossRefPubMedCentralPubMedGoogle Scholar
  53. 53.
    Wolf W, Kilic A, Schrul B, Lorenz H, Schwappach B, Seedorf M (2012) Yeast Ist2 recruits the endoplasmic reticulum to the plasma membrane and creates a ribosome-free membrane microcompartment. PLoS ONE 7:e39703CrossRefPubMedCentralPubMedGoogle Scholar
  54. 54.
    Yang H, Kim A, David T, Palmer D, Jin T, Tien J, Huang F, Cheng T, Coughlin SR, Jan YN, Jan LY (2012) TMEM16F forms a Ca(2+)-activated cation channel required for lipid scrambling in platelets during blood coagulation. Cell 151:111–122CrossRefPubMedCentralPubMedGoogle Scholar
  55. 55.
    Yang YD, Cho H, Koo JY, Tak MH, Cho Y, Shim WS, Park SP, Lee J, Lee B, Kim BM, Raouf R, Shin YK, Oh U (2008) TMEM16A confers receptor-activated calcium-dependent chloride conductance. Nature 455:1210–1215CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Rainer Schreiber
    • 1
  • Diana Faria
    • 1
  • Boris V. Skryabin
    • 2
    • 6
  • Podchanart Wanitchakool
    • 1
  • Jason R. Rock
    • 3
    • 4
    • 5
  • Karl Kunzelmann
    • 1
    Email author
  1. 1.Institut für PhysiologieUniversität RegensburgRegensburgGermany
  2. 2.Institut für Experimentelle Pathologie (ZMBE)Westfälischen Wilhelms-Universität MünsterMünsterGermany
  3. 3.Department of AnatomyUniversity California, San FranciscoSan FranciscoUSA
  4. 4.Department of MedicineUniversity California, San FranciscoSan FranciscoUSA
  5. 5.Cardiovascular Research InstituteUniversity California, San FranciscoSan FranciscoUSA
  6. 6.Interdisciplinary Center for Clinical Research (IZKF)University of MünsterMünsterGermany

Personalised recommendations