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Cholinergic signalling-regulated KV7.5 currents are expressed in colonic ICC-IM but not ICC-MP

  • Ion channels, receptors and transporters
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Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

Interstitial cells of Cajal (ICC) and the enteric nervous system orchestrate the various rhythmic motor patterns of the colon. Excitation of ICC may evoke stimulus-dependent pacemaker activity and will therefore have a profound effect on colonic motility. The objective of the present study was to evaluate the potential role of K+ channels in the regulation of ICC excitability. We employed the cell-attached patch clamp technique to assess single channel activity from mouse colon ICC, immunohistochemistry to determine ICC K+ channel expression and single cell RT-PCR to identify K+ channel RNA. Single channel activity revealed voltage-sensitive K+ channels, which were blocked by the KV7 blocker XE991 (n = 8), which also evoked inward maxi channel activity. Muscarinic acetylcholine receptor stimulation with carbachol inhibited K+ channel activity (n = 8). The single channel conductance was 3.4 ± 0.1 pS (n = 8), but with high extracellular K+, it was 18.1 ± 0.6 pS (n = 22). Single cell RT-PCR revealed Ano1-positive ICC that were positive for KV7.5. Double immunohistochemical staining of colons for c-Kit and KV7.5 in situ revealed that intramuscular ICC (ICC-IM), but not ICC associated with the myenteric plexus (ICC-MP), were positive for KV7.5. It also revealed dense cholinergic innervation of ICC-IM. ICC-IM and ICC-MP networks were found to be connected. We propose that the pacemaker network in the colon consists of both ICC-MP and ICC-IM and that one way of exciting this network is via cholinergic KV7.5 channel inhibition in ICC-IM.

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Abbreviations

GI:

Gastrointestinal

ICC:

Interstitial cells of Cajal

ICC-MP:

Interstitial cells of Cajal associated with the myenteric plexus

ICC-IM:

Intramuscular interstitial cells of Cajal

ICC-SMP:

Interstitial cells of Cajal associated with the submuscular plexus

KV7.5:

Voltage-gated K+ channel 7.5

mAChRs:

Muscarinic acetylcholine receptors

rrmp:

Relative to resting membrane potential

VAChT:

Vesicular acetylcholine transporter

References

  1. Anderson UA, Carson C, McCloskey KD (2009) KCNQ currents and their contribution to resting membrane potential and the excitability of interstitial cells of Cajal from the guinea pig bladder. J Urol 182:330–336

    Google Scholar 

  2. Bal M, Zhang J, Zaika O, Hernandez CC, Shapiro MS (2008) Homomeric and heteromeric assembly of KCNQ (Kv7) K+ channels assayed by total internal reflection fluorescence/fluorescence resonance energy transfer and patch clamp analysis. J Biol Chem 283:30668–30676

    Google Scholar 

  3. Block BM, Jones SW (1997) Delayed rectifier current of bullfrog sympathetic neurons: ion-ion competition, asymmetrical block and effects of ions on gating. J Physiol 499(Pt 2):403–416

    Google Scholar 

  4. Brown DA, Adams PR (1980) Muscarinic suppression of a novel voltage-sensitive K+ current in a vertebrate neurone. Nature 283:673–676

    Google Scholar 

  5. Brown DA, Hughes SA, Marsh SJ, Tinker A (2007) Regulation of M(Kv7.2/7.3) channels in neurons by PIP(2) and products of PIP(2) hydrolysis: significance for receptor-mediated inhibition. J Physiol 582:917–925

    Google Scholar 

  6. Brown DA, Passmore GM (2009) Neural KCNQ (Kv7) channels. Br J Pharmacol 156:1185–1195

    Google Scholar 

  7. Chen H, Ordog T, Chen J, Young DL, Bardsley MR, Redelman D, Ward SM, Sanders KM (2007) Differential gene expression in functional classes of interstitial cells of Cajal in murine small intestine. Physiol Genomics 31:492–509

    Google Scholar 

  8. Chen JH, Zhang Q, Yu Y, Li K, Liao H, Jiang L, Hong L, Du X, Hu X, Chen S, Yin S, Gao Q, Yin X, Luo H, Huizinga JD (2013) Neurogenic and myogenic properties of pan-colonic motor patterns and their spatiotemporal organization in rats. PLoS One 8:e60474

    Google Scholar 

  9. Costa M, Dodds KN, Wiklendt L, Spencer NJ, Brookes SJ, Dinning PG (2013) Neurogenic and myogenic motor activity in the colon of the guinea-pig, mouse, rabbit and rat. Am J Physiol Gastrointest Liver Physiol 305:G749–G759

    Google Scholar 

  10. Delmas P, Brown DA (2005) Pathways modulating neural KCNQ/M (Kv7) potassium channels. Nat Rev Neurosci 6:850–862

    Google Scholar 

  11. Edwards FR, Hirst GD (2006) An electrical analysis of slow wave propagation in the guinea-pig gastric antrum. J Physiol 571:179–189

    Google Scholar 

  12. Faussone Pellegrini MS, Cortesini C, Romagnoli P (1977) Ultrastructure of the tunica muscularis of the cardial portion of the human esophagus and stomach, with special reference to the so-called Cajal's interstitial cells. Arch Ital Anat Embriol 82:157–177

    Google Scholar 

  13. Hennig GW, Hirst GD, Park KJ, Smith CB, Sanders KM, Ward SM, Smith TK (2004) Propagation of pacemaker activity in the guinea-pig antrum. J Physiol 556:585–599

    Google Scholar 

  14. Hirst GD, Beckett EA, Sanders KM, Ward SM (2002) Regional variation in contribution of myenteric and intramuscular interstitial cells of Cajal to generation of slow waves in mouse gastric antrum. J Physiol 540:1003–1012

    Google Scholar 

  15. Hirst GD, Dickens EJ, Edwards FR (2002) Pacemaker shift in the gastric antrum of guinea-pigs produced by excitatory vagal stimulation involves intramuscular interstitial cells. J Physiol 541:917–928

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Hirst GD, Garcia-Londono AP, Edwards FR (2006) Propagation of slow waves in the guinea-pig gastric antrum. J Physiol 571:165–177

    Article  PubMed Central  PubMed  Google Scholar 

  17. Huizinga JD and Chen JH (2014) Interstitial cells of Cajal: update on clinical and basic science. Curr Gastroenterol Rep 16:363–374

    Google Scholar 

  18. Huizinga JD, Martz S, Gil V, Wang XY, Jimenez M, Parsons S (2011) Two independent networks of interstitial cells of Cajal work cooperatively with the enteric nervous system to create colonic motor patterns. Front Neurosci 5:93

    Article  PubMed Central  PubMed  Google Scholar 

  19. Huizinga JD, Thuneberg L, Kluppel M, Malysz J, Mikkelsen HB, Bernstein A (1995) W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373:347–349

    Article  CAS  PubMed  Google Scholar 

  20. Huizinga JD, Zhu Y, Ye J, Molleman A (2002) High-conductance chloride channels generate pacemaker currents in interstitial cells of Cajal. Gastroenterology 123:1627–1636

    Article  CAS  PubMed  Google Scholar 

  21. Ipavec V, Martire M, Barrese V, Taglialatela M, Curro D (2011) KV7 channels regulate muscle tone and nonadrenergic noncholinergic relaxation of the rat gastric fundus. Pharmacol Res 64:397–409

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Jepps TA, Greenwood IA, Moffatt JD, Sanders KM, Ohya S (2009) Molecular and functional characterization of Kv7 K+ channel in murine gastrointestinal smooth muscles. Am J Physiol Gastrointest Liver Physiol 297:G107–G115

    CAS  PubMed Central  PubMed  Google Scholar 

  23. Komuro T (2006) Structure and organization of interstitial cells of Cajal in the gastrointestinal tract. J Physiol 576:653–658

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Lerche C, Scherer CR, Seebohm G, Derst C, Wei AD, Busch AE, Steinmeyer K (2000) Molecular cloning and functional expression of KCNQ5, a potassium channel subunit that may contribute to neuronal M-current diversity. J Biol Chem 275:22395–22400

    Article  CAS  PubMed  Google Scholar 

  25. Li Y, Gamper N, Shapiro MS (2004) Single-channel analysis of KCNQ K+ channels reveals the mechanism of augmentation by a cysteine-modifying reagent. J Neurosci 24:5079–5090

    Article  CAS  PubMed  Google Scholar 

  26. Linley JE, Pettinger L, Huang D, Gamper N (2012) M channel enhancers and physiological M channel block. J Physiol 590:793–807

    CAS  PubMed Central  PubMed  Google Scholar 

  27. Liu LW, Huizinga JD (1993) Electrical coupling of circular muscle to longitudinal muscle and interstitial cells of Cajal in canine colon. J Physiol 470:445–461

    CAS  PubMed Central  PubMed  Google Scholar 

  28. Molleman A (2003) Patch clamping an introductory guide to patch clamp electrophysiology. Wiley, New York

    Google Scholar 

  29. Parsons SP, Huizinga JD (2010) Transient outward potassium current in ICC. Am J Physiol Gastrointest Liver Physiol 298:G456–G466

    CAS  PubMed  Google Scholar 

  30. Parsons SP, Kunze WA, Huizinga JD (2012) Maxi-channels recorded in situ from ICC and pericytes associated with the mouse myenteric plexus. Am J Physiol Cell Physiol 302:C1055–C1069

    CAS  PubMed  Google Scholar 

  31. Parsons SP, Sanders KM (2008) An outwardly rectifying and deactivating chloride channel expressed by interstitial cells of Cajal from the murine small intestine. J Membr Biol 221:123–132

    Article  CAS  PubMed  Google Scholar 

  32. Pluja L, Alberti E, Fernandez E, Mikkelsen HB, Thuneberg L, Jimenez M (2001) Evidence supporting presence of two pacemakers in rat colon. Am J Physiol Gastrointest Liver Physiol 281:G255–G266

    CAS  PubMed  Google Scholar 

  33. Rae MG, Fleming N, McGregor DB, Sanders KM, Keef KD (1998) Control of motility patterns in the human colonic circular muscle layer by pacemaker activity. J Physiol 510(Pt 1):309–320

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Roura-Ferrer M, Etxebarria A, Sole L, Oliveras A, Comes N, Villarroel A, Felipe A (2009) Functional implications of KCNE subunit expression for the Kv7.5 (KCNQ5) channel. Cell Physiol Biochem 24:325–334

    Article  CAS  PubMed  Google Scholar 

  35. Rumessen JJ, Thuneberg L, Mikkelsen HB (1982) Plexus muscularis profundus and associated interstitial cells. II. Ultrastructural studies of mouse small intestine. Anat Rec 203:129–146

    Article  CAS  PubMed  Google Scholar 

  36. Sanders KM, Koh SD, Ward SM (2006) Interstitial cells of Cajal as pacemakers in the gastrointestinal tract. Annu Rev Physiol 68:307–343

    Article  CAS  PubMed  Google Scholar 

  37. Schroeder BC, Hechenberger M, Weinreich F, Kubisch C, Jentsch TJ (2000) KCNQ5, a novel potassium channel broadly expressed in brain, mediates M-type currents. J Biol Chem 275:24089–24095

    Article  CAS  PubMed  Google Scholar 

  38. Shibasaki T (1987) Conductance and kinetics of delayed rectifier potassium channels in nodal cells of the rabbit heart. J Physiol 387:227–250

    CAS  PubMed Central  PubMed  Google Scholar 

  39. Smith TK, Reed JB, Sanders KM (1987) Interaction of two electrical pacemakers in muscularis of canine proximal colon. Am J Physiol 252:C290–C299

    CAS  PubMed  Google Scholar 

  40. Thomsen L, Robinson TL, Lee JC, Farraway LA, Hughes MJ, Andrews DW, Huizinga JD (1998) Interstitial cells of Cajal generate a rhythmic pacemaker current. Nat Med 4:848–851

    Article  CAS  PubMed  Google Scholar 

  41. Thuneberg L (1982) Interstitial cells of Cajal: intestinal pacemaker cells? Adv Anat Embryol Cell Biol 71:1–130

    Article  CAS  PubMed  Google Scholar 

  42. Thuneberg L (1999) One hundred years of interstitial cells of Cajal. Microsc Res Tech 47:223–238

    Article  CAS  PubMed  Google Scholar 

  43. van Breemen C, Chen Q, Laher I (1995) Superficial buffer barrier function of smooth muscle sarcoplasmic reticulum. Trends Pharmacol Sci 16:98–105

    Article  PubMed  Google Scholar 

  44. Wang B, Kunze WA, Zhu Y, Huizinga JD (2008) In situ recording from gut pacemaker cells. Pflugers Arch 457:243–251

    Article  CAS  PubMed  Google Scholar 

  45. Wright G, Parsons SP, Huizinga JD (2012) Colonic interstitial cells of Cajal associated with the myenteric plexus harbour ion channels regulated by excitatory neurotransmitters. Neurogastroenterol Motil 24(S2):156

    Google Scholar 

  46. Wright GW, Parsons SP, Huizinga JD (2012) Ca(2+) sensitivity of the maxi chloride channel in interstitial cells of Cajal. Neurogastroenterol Motil 24:e221–e234

    Article  CAS  PubMed  Google Scholar 

  47. Yoneda S, Fukui H, Takaki M (2004) Pacemaker activity from submucosal interstitial cells of Cajal drives high-frequency and low-amplitude circular muscle contractions in the mouse proximal colon. Neurogastroenterol Motil 16:621–627

    Article  CAS  PubMed  Google Scholar 

  48. Zhong XZ, Harhun MI, Olesen SP, Ohya S, Moffatt JD, Cole WC, Greenwood IA (2010) Participation of KCNQ (Kv7) potassium channels in myogenic control of cerebral arterial diameter. J Physiol 588:3277–3293

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  49. Zhu MH, Kim TW, Ro S, Yan W, Ward SM, Koh SD, Sanders KM (2009) A Ca(2+)-activated Cl(−) conductance in interstitial cells of Cajal linked to slow wave currents and pacemaker activity. J Physiol 587:4905–4918

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the Natural Sciences and Engineering Research Council (NSERC) operating grant #386877 to JH. GW was supported by both a doctoral NSERC Postgraduate Scholarship and an Ontario Graduate Scholarship. Additional support was obtained from the Canadian Institutes of Health Research #MOP-12874 to JH. RLV was supported by CONACYT (Consejo Nacional de Ciencia y Tecnología, México, scholarship 290618). Some of the data in this manuscript were previously published in abstract form [45]. We thankfully acknowledge Dr. Waliul Khan for the use of his thermal cycler and documentation system. We also acknowledge Drs. Luke Janssen and Wolfgang Kunze for their helpful discussions on analysis.

Ethical standards

The experiments described herein were performed in accordance with the guidelines of the Canadian Council on Animal Care (CCAC) and received approval from the McMaster University Animal Research Ethics Board (AREB).

Conflicts of interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Farncombe Family Digestive Health Research Institute, Department of Medicine, McMaster University, HSC-3N8, 1200 Main Street West, Hamilton, ON, L8N 3Z5, Canada

    George W. J. Wright, Sean P. Parsons, Raúl Loera-Valencia, Xuan-Yu Wang & Jan D. Huizinga

  2. IPICYT (Instituto Potosino de Investigación Científica y Technológica), San Luis Potosí, SLP, Mexico

    Raúl Loera-Valencia & Carlos Barajas-López

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  1. George W. J. Wright
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  6. Jan D. Huizinga
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Correspondence to George W. J. Wright or Jan D. Huizinga.

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Fig. 1

Double labeling of c-Kit (green) and KV7.2/7.3/7.4 (red) in mouse colon. Top three groups of figures were from cross sections, bottom group from musculature wholemount preparation. Enteric nerves at the level of myenteric plexus (MP) were positive for KV7.2 and KV7.4. Enteric nerves and smooth muscles in the longitudinal muscle (LM) layer were positive for KV7.3. No co-localization was found between KV7.2/7.3/7.4 and c-Kit. CM: circular muscle (JPEG 221 kb)

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Wright, G.W.J., Parsons, S.P., Loera-Valencia, R. et al. Cholinergic signalling-regulated KV7.5 currents are expressed in colonic ICC-IM but not ICC-MP. Pflugers Arch - Eur J Physiol 466, 1805–1818 (2014). https://doi.org/10.1007/s00424-013-1425-7

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