Skip to main content

Advertisement

SpringerLink
Log in

Functional link between muscarinic receptors and large-conductance Ca2+-activated K+ channels in freshly isolated human detrusor smooth muscle cells

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

Abstract

Activation of muscarinic acetylcholine receptors (mAChRs) constitutes the primary mechanism for enhancing excitability and contractility of human detrusor smooth muscle (DSM). Since the large-conductance Ca2+-activated K+ (KCa1.1) channels are key regulators of human DSM function, we investigated whether mAChR activation increases human DSM excitability by inhibiting KCa1.1 channels. We used the mAChR agonist, carbachol, to determine the changes in KCa1.1 channel activity upon mAChR activation in freshly isolated human DSM cells obtained from open bladder surgeries using the perforated whole cell and single KCa1.1 channel patch-clamp recordings. Human DSM cells were collected from 29 patients (23 males and 6 females, average age of 65.9 ± 1.5 years). Carbachol inhibited the amplitude and frequency of KCa1.1 channel-mediated spontaneous transient outward currents and spontaneous transient hyperpolarizations, which are triggered by the release of Ca2+ from ryanodine receptors. Carbachol also caused membrane potential depolarization, which was not observed in the presence of iberiotoxin, a KCa1.1 channel inhibitor, indicating the critical role of the KCa1.1 channels. The potential direct carbachol effects on KCa1.1 channels were examined under conditions of removing the major cellular Ca2+ sources for KCa1.1 channel activation with pharmacological inhibitors (thapsigargin, ryanodine, and nifedipine). In the presence of these inhibitors, carbachol did not affect the single KCa1.1 channel open probability and mean KCa1.1 channel conductance (cell-attached configuration) or depolarization-induced whole cell steady-state KCa1.1 currents. The data support the concept that mAChR activation triggers indirect functional KCa1.1 channel inhibition mediated by intracellular Ca2+, thus increasing the excitability in human DSM cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Abrams P, Cardozo L, Fall M, Griffiths D, Rosier P, Ulmsten U, van Kerrebroeck P, Victor A, Wein A (2002) The standardisation of terminology of lower urinary tract function: report from the Standardisation Sub-committee of the International Continence Society. Am J Obstet Gynecol 187(1):116–126

    Article  PubMed  Google Scholar 

  2. Andersson KE, Arner A (2004) Urinary bladder contraction and relaxation: physiology and pathophysiology. Physiol Rev 84(3):935–986

    Article  CAS  PubMed  Google Scholar 

  3. Andersson KE, Holmquist F, Fovaeus M, Hedlund H, Sundler R (1991) Muscarinic receptor stimulation of phosphoinositide hydrolysis in the human isolated urinary bladder. J Urol 146(4):1156–1159

    CAS  PubMed  Google Scholar 

  4. Bayguinov O, Hagen B, Sanders KM (2001) Muscarinic stimulation increases basal Ca2+ and inhibits spontaneous Ca2+ transients in murine colonic myocytes. Am J Physiol Cell Physiol 280(3):C689–700

    CAS  PubMed  Google Scholar 

  5. Brown SM, Bentcheva-Petkova LM, Liu L, Hristov KL, Chen M, Kellett WF, Meredith AL, Aldrich RW, Nelson MT, Petkov GV (2008) Beta-adrenergic relaxation of mouse urinary bladder smooth muscle in the absence of large-conductance Ca2+-activated K+ channel. Am J Physiol Renal Physiol 295(4):F1149–1157

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Caulfield MP, Birdsall NJ (1998) International Union of Pharmacology. XVII. Classification of muscarinic acetylcholine receptors. Pharmacol Rev 50(2):279–290

    CAS  PubMed  Google Scholar 

  7. Chess-Williams R, Chapple CR, Yamanishi T, Yasuda K, Sellers DJ (2001) The minor population of M3-receptors mediate contraction of human detrusor muscle in vitro. J Auton Pharmacol 21(5–6):243–248

    Article  CAS  PubMed  Google Scholar 

  8. Cox DH, Aldrich RW (2000) Role of the beta1 subunit in large-conductance Ca2+-activated K+ channel gating energetics. Mechanisms of enhanced Ca2+ sensitivity. J Gen Physiol 116(3):411–432

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Epstein LB, Goldberg RP (2005) The overactive bladder and quality of life. Int J Fertil Womens Med 50(1):30–36

    PubMed  Google Scholar 

  10. Hashitani H, Brading AF (2003) Electrical properties of detrusor smooth muscles from the pig and human urinary bladder. Br J Pharmacol 140(1):146–158

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Hashitani H, Fukuta H, Takano H, Klemm MF, Suzuki H (2001) Origin and propagation of spontaneous excitation in smooth muscle of the guinea-pig urinary bladder. J Physiol 530(Pt 2):273–286

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Herrera GM, Heppner TJ, Nelson MT (2001) Voltage dependence of the coupling of Ca2+ sparks to BKCa channels in urinary bladder smooth muscle. Am J Physiol Cell Physiol 280(3):C481–490

    CAS  PubMed  Google Scholar 

  13. Herrera GM, Nelson MT (2002) Differential regulation of SK and BK channels by Ca2+ signals from Ca2+ channels and ryanodine receptors in guinea-pig urinary bladder myocytes. J Physiol 541(Pt 2):483–492

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Hotta S, Morimura K, Ohya S, Muraki K, Takeshima H, Imaizumi Y (2007) Ryanodine receptor type 2 deficiency changes excitation-contraction coupling and membrane potential in urinary bladder smooth muscle. J Physiol 582(Pt 2):489–506

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Hristov KL, Chen M, Kellett WF, Rovner ES, Petkov GV (2011) Large-conductance voltage- and Ca2+-activated K+ channels regulate human detrusor smooth muscle function. Am J Physiol Cell Physiol 301(4):C903–912

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Hristov KL, Parajuli SP, Soder RP, Cheng Q, Rovner ES, Petkov GV (2012) Suppression of human detrusor smooth muscle excitability and contractility via pharmacological activation of large conductance Ca2+-activated K+ channels. Am J Physiol Cell Physiol 302(11):C1632–1641

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Hristov KL, Smith AC, Parajuli SP, Malysz J, Petkov GV (2014) Large-conductance voltage- and Ca2+-activated K+ channel regulation by protein kinase C in guinea pig urinary bladder smooth muscle. Am J Physiol Cell Physiol 306(5):C460–470

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Jaggar JH, Porter VA, Lederer WJ, Nelson MT (2000) Calcium sparks in smooth muscle. Am J Physiol Cell Physiol 278(2):C235–256

    CAS  PubMed  Google Scholar 

  19. Malysz J, Rovner ES, Petkov GV (2013) Single-channel biophysical and pharmacological characterizations of native human large-conductance calcium-activated potassium channels in freshly isolated detrusor smooth muscle cells. Pflugers Arch 465(7):965–975

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Morimura K, Ohi Y, Yamamura H, Ohya S, Muraki K, Imaizumi Y (2006) Two-step Ca2+ intracellular release underlies excitation-contraction coupling in mouse urinary bladder myocytes. Am J Physiol Cell Physiol 290(2):C388–403

    Article  CAS  PubMed  Google Scholar 

  21. Nakamura T, Kimura J, Yamaguchi O (2002) Muscarinic M2 receptors inhibit Ca2+-activated K+ channels in rat bladder smooth muscle. Int J Urol 9(12):689–696

    Article  CAS  PubMed  Google Scholar 

  22. Orio P, Latorre R (2005) Differential effects of b1 and b2 subunits on BK channel activity. J Gen Physiol 125(4):395–411

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Parajuli SP, Petkov GV (2013) Activation of muscarinic M3 receptors inhibits large-conductance voltage- and Ca2+-activated K+ channels in rat urinary bladder smooth muscle cells. Am J Physiol Cell Physiol 305(2):C207–214

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Perez GJ, Bonev AD, Patlak JB, Nelson MT (1999) Functional coupling of ryanodine receptors to KCa channels in smooth muscle cells from rat cerebral arteries. J Gen Physiol 113(2):229–238

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Petkov GV (2012) Role of potassium ion channels in detrusor smooth muscle function and dysfunction. Nat Rev Urol 9(1):30–40

    Article  CAS  Google Scholar 

  26. Petkov GV, Heppner TJ, Bonev AD, Herrera GM, Nelson MT (2001) Low levels of KATP channel activation decrease excitability and contractility of urinary bladder. Am J Physiol Regul Integr Comp Physiol 280(5):R1427–1433

    CAS  PubMed  Google Scholar 

  27. Petkov GV, Nelson MT (2005) Differential regulation of Ca2+-activated K+ channels by beta-adrenoceptors in guinea pig urinary bladder smooth muscle. Am J Physiol Cell Physiol 288(6):C1255–1263

    Article  CAS  PubMed  Google Scholar 

  28. Robbins J (1993) Agonist-induced inhibition of inositol-trisphosphate-activated IK(Ca) in NG108-15 neuroblastoma hybrid cells. Pflugers Arch 422(4):364–370

    Article  CAS  PubMed  Google Scholar 

  29. Schneider T, Fetscher C, Krege S, Michel MC (2004) Signal transduction underlying carbachol-induced contraction of human urinary bladder. J Pharmacol Exp Ther 309(3):1148–1153

    Article  CAS  PubMed  Google Scholar 

  30. Vaithianathan T, Bukiya A, Liu J, Liu P, Asuncion-Chin M, Fan Z, Dopico A (2008) Direct regulation of BK channels by phosphatidylinositol 4,5-bisphosphate as a novel signaling pathway. J Gen Physiol 132(1):13–28

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Wu C, Bayliss M, Newgreen D, Mundy AR, Fry CH (1999) A comparison of the mode of action of ATP and carbachol on isolated human detrusor smooth muscle. J Urol 162(5):1840–1847

    Article  CAS  PubMed  Google Scholar 

  32. Wuest M, Hiller N, Braeter M, Hakenberg OW, Wirth MP, Ravens U (2007) Contribution of Ca2+ influx to carbachol-induced detrusor contraction is different in human urinary bladder compared to pig and mouse. Eur J Pharmacol 565(1–3):180–189

    Article  CAS  PubMed  Google Scholar 

  33. Xin W, Cheng Q, Soder RP, Rovner ES, Petkov GV (2012) Constitutively active phosphodiesterase activity regulates urinary bladder smooth muscle function: critical role of KCa1.1 channel. Am J Physiol Renal Physiol 303(9):F1300–1306

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Xin W, Soder RP, Cheng Q, Rovner ES, Petkov GV (2012) Selective inhibition of phosphodiesterase 1 relaxes urinary bladder smooth muscle: role for ryanodine receptor-mediated BK channel activation. Am J Physiol Cell Physiol 303(10):C1079–1089

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Yamaguchi O, Shishido K, Tamura K, Ogawa T, Fujimura T, Ohtsuka M (1996) Evaluation of mRNAs encoding muscarinic receptor subtypes in human detrusor muscle. J Urol 156(3):1208–1213

    Article  CAS  PubMed  Google Scholar 

  36. Zhou XB, Wulfsen I, Lutz S, Utku E, Sausbier U, Ruth P, Wieland T, Korth M (2008) M2 muscarinic receptors induce airway smooth muscle activation via a dual, Gbetagamma-mediated inhibition of large conductance Ca2+-activated K+ channel activity. J Biol Chem 283(30):21036–21044

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to thank the Medical University of South Carolina (MUSC) Department of Urology staff surgeons Drs. Thomas Keane, Harry Clarke, Stephen Savage, Ross Rames, Jonathan Picard, Sandip Prasad, and Ahmed M. El-Zawahry as well as the MUSC Urology Residents Drs. Matthew Young, Erin Burns, Vaughan Taylor, Samuel Walker Nickles, Justin Ellett, Ryan Levey, Austin Younger, and Nima Baradaran for their help with human tissue collection. We would like to thank Drs. Wenkuan Xin and Ning Li, Mr. Aaron Provence, Mr. Vitor Fernandes, and Ms. Amy Smith for the critical evaluation of the manuscript.

Grants

This study was supported by a grant from the National Institutes of Health R01 DK084284 to Georgi V. Petkov and by a fellowship from the Urology Care Foundation Research Scholars Program and the Allergan Foundation to Shankar P. Parajuli.

Conflicts of interest

The authors declare no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Drug Discovery and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, Coker Life Sciences Building, Room 609D, 715 Sumter St, Columbia, SC, 29208, USA

    Shankar P. Parajuli, Kiril L. Hristov, Qiuping Cheng, John Malysz & Georgi V. Petkov

  2. Medical University of South Carolina, Charleston, SC, 29425, USA

    Eric S. Rovner & Georgi V. Petkov

Authors
  1. Shankar P. Parajuli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kiril L. Hristov
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qiuping Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John Malysz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Eric S. Rovner
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Georgi V. Petkov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Georgi V. Petkov.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Parajuli, S.P., Hristov, K.L., Cheng, Q. et al. Functional link between muscarinic receptors and large-conductance Ca2+-activated K+ channels in freshly isolated human detrusor smooth muscle cells. Pflugers Arch - Eur J Physiol 467, 665–675 (2015). https://doi.org/10.1007/s00424-014-1537-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-014-1537-8

Keywords

Search

Navigation