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Effects of intracellular pH and Ca2+ on the activity of stretch-sensitive cation channels in leech neurons

  • Ion Channels, Transporters
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Abstract

The effects of intracellular pH and calcium on the activity of the leech mechanosensitive cation channels have been studied. These channels exhibited two activity modes denoted as spike-like (SL) and multiconductance (MC). In the absence of mechanical stimulation, acidification of the intracellular side of membrane patches from 7.2 to 6.2 reversibly increased the mean channel open time as well as the opening frequency in the SL mode. Channels in MC mode were activated by a pHi reduction from 7.2 to 6.2, but were inhibited at pHi 5.5. Unlike MC mode, SL mode was strongly activated by intracellular Ca2+. Fura-2 imaging experiments showed that intracellular calcium was induced to increase by hypotonic cell swelling. The major component of this response did not require extracellular calcium. A component of the swelling-induced calcium response was sensitive to blockers of stretch-sensitive cation channels. The results indicate that the two activity modes of mechanosensitive channels of leech neurons respond differently to changes of intracellular pH and calcium. The sensitivity of the channel to micromolar concentrations of internal free calcium, along with its permeability to this ion, is consistent with a role in the amplification of mechanically induced Ca2+ signals in leech neurons.

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References

  1. Blackshaw SE, Nicholls JG, Parnas I (1982) Expanded receptive fields of cutaneous mechanoreceptor cells after single neurone deletion in leech central nervous system. J Physiol 326:261–268

    PubMed  CAS  Google Scholar 

  2. Calabrese B, Manzi S, Pellegrini M, Pellegrino M (1999) Stretch-activated cation channels of leech neurons: characterization and role in neurite outgrowth. Eur J Neurosci 11:2275–2284

    Article  PubMed  CAS  Google Scholar 

  3. Clapham DE (2003) TRP channels as cellular sensors. Nature 426:517–524

    Article  PubMed  CAS  Google Scholar 

  4. Corey DP, García-Añoveros J, Holt JR, Kwan KY, Lin S, Vollrath MA, Amalfitano A, Cheung EL, Derfler BH, Duggan A, Géléoc GS, Gray PA, Hoffman MP, Rehm HL, Tamasauskas D, Zhang D (2004) TRPA1 is a candidate for the mechanosensitive transduction channel of vertebrate hair cells. Nature 432:723–730

    Article  PubMed  CAS  Google Scholar 

  5. Corey DP, Stevens CF (1983) Science and technology of patch-recordings electrodes. In: Sakmann B, Neher E (eds) Single channel recording. Plenum, New York, pp 53–68

    Google Scholar 

  6. Fleig A, Penner R (2004) The TRPM ion channel subfamily: molecular, biophysical and functional features. Trends Pharmacol Sci 25:633–639

    Article  PubMed  CAS  Google Scholar 

  7. Fuchs PA, Nicholls JG, Ready D (1981) Membrane properties and selective connexions of identified leech neurones in culture. J Physiol 316:203–223

    PubMed  CAS  Google Scholar 

  8. Gomez T (2005) Channels for pathfinding. Nature 434:835–838

    Article  PubMed  CAS  Google Scholar 

  9. Gomez TM, Spitzer NC (1999) In vivo regulation of axon extension and pathfinding by growth-cone calcium transients. Nature 397:350–355

    Article  PubMed  CAS  Google Scholar 

  10. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    PubMed  CAS  Google Scholar 

  11. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100

    Article  PubMed  CAS  Google Scholar 

  12. Hamill OP, McBride DW Jr (1996) The pharmacology of mechanogated membrane ion channels. Pharmacol Rev 48:231–252

    PubMed  CAS  Google Scholar 

  13. Hamill OP, McBride DW Jr (1997) Induced membrane hypo/hyper-mechanosensitivity: a limitation of patch-clamp recording. Annu Rev Physiol 59:621–631

    Article  PubMed  CAS  Google Scholar 

  14. Honoré E, Maingret F, Lazdunski M, Patel AJ (2002) An intracellular proton sensor commands lipid- and mechano-gating of the K+ channel TREK-1. EMBO J 21:2968–2976

    Article  PubMed  Google Scholar 

  15. Hurwitz CG, Segal AS (2001) Application of pressure steps to mechanosensitive channels in membrane patches: a simple, economical, and fast system. Pflugers Arch 442:150–156

    Article  PubMed  CAS  Google Scholar 

  16. Ishikawa T, Marunaka Y, Rotin D (1998) Electrophysiological characterization of the rat epithelial Na+ channel (rENaC) expressed in MDCK cells. Effects of Na+ and Ca2+. J Gen Physiol 111:825–846

    Article  PubMed  CAS  Google Scholar 

  17. Li Y, Jia YC, Cui K, Li N, Zheng ZY, Wang Y, Yuan X (2005) Essential role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor. Nature 434:894–898

    Article  PubMed  CAS  Google Scholar 

  18. Lin S, Corey DP (2005) TRP channels in mechanosensation. Curr Opin Neurobiol 15:350–357

    Article  PubMed  CAS  Google Scholar 

  19. Maingret F, Patel AJ, Lesage F, Lazdunski M, Honoré E (1999) Mechano- or acid stimulation, two interactive modes of activation of the TREK-1 potassium channel. J Biol Chem 274:26691–26696

    Article  PubMed  CAS  Google Scholar 

  20. Maroto R, Raso A, Wood TG, Kurosky A, Martinac B, Hamill OP (2005) TRPC1 forms the stretch-activated cation channel in vertebrate cells. Nat Cell Biol 7:179–185

    Article  PubMed  CAS  Google Scholar 

  21. Martinac B (2004) Mechanosensitive ion channels: molecules of mechanotransduction. J Cell Sci 117:2449–2460

    Article  PubMed  CAS  Google Scholar 

  22. Menconi MC, Pellegrini M, Pellegrino M (2001) Voltage-induced activation of mechanosensitive cation channels of leech neurons. J Membr Biol 180:65–72

    Article  PubMed  CAS  Google Scholar 

  23. Nagata K, Duggan A, Kumar G, García-Añoveros J (2005) Nociceptor and hair cell transducer properties of TRPA1, a channel for pain and hearing. J Neurosci 25:4052–4061

    Article  PubMed  CAS  Google Scholar 

  24. Nilius B, Voets T (2005) TRP channels: a TR(I)P through a world of multifunctional cation channels. Pflugers Arch 451:1–10

    Article  PubMed  CAS  Google Scholar 

  25. O’Hagan R, Chalfie M, Goodman MB (2005) The MEC-4 DEG/ENaC channel of Caenorhabditis elegans touch receptor neurons transduces mechanical signals. Nat Neurosci 8:43–50

    Article  PubMed  CAS  Google Scholar 

  26. O’Neil RG, Heller S (2005) The mechanosensitive nature of TRPV channels. Pflugers Arch 451:193–203

    Article  PubMed  CAS  Google Scholar 

  27. Parekh AB, Putney JW Jr (2005) Store-operated calcium channels. Physiol Rev 85:757–810

    Article  PubMed  CAS  Google Scholar 

  28. Patel AJ, Honoré E (2001) Properties and modulation of mammalian 2P domain K+ channels. Trends Neurosci 24:339–346

    Article  PubMed  CAS  Google Scholar 

  29. Pellegrini M, Menconi MC, Pellegrino M (2001) Stretch-activated cation channels of leech neurons exhibit two activity modes. Eur J Neurosci 13:503–511

    Article  PubMed  CAS  Google Scholar 

  30. Pellegrino M, Pellegrini M, Simoni A, Gargini C (1990) Stretch-activated cation channels with large unitary conductance in leech central neurons. Brain Res 525:322–326

    Article  PubMed  CAS  Google Scholar 

  31. Shim S, Goh EL, Ge S, Sailor K, Yuan JP, Roderick HL, Bootman MD, Worley PF, Song H, Ming G (2005) XTRPC1-dependent chemotropic guidance of neuronal growth cones. Nat Neurosci 8:730–735

    Article  PubMed  CAS  Google Scholar 

  32. Spassova MA, Soboloff J, He L, Hewavitharana T, Xu W, Venkatachalam K, van Rossum DB, Patterson RL, Gill DL (2004) Calcium entry mediated by SOCs and TRP channels: variations and enigma. Biochim Biophys Acta 1742:9–20

    Article  PubMed  CAS  Google Scholar 

  33. Strassmaier M, Gillespie PG (2002) The hair cell’s transduction channel. Curr Opin Neurobiol 12:380–386

    Article  PubMed  CAS  Google Scholar 

  34. Sukharev S, Anishkin A (2004) Mechanosensitive channels: what can we learn from “simple” model systems? Trends Neurosci 27:345–351

    Article  PubMed  CAS  Google Scholar 

  35. Tan JH, Liu W, Saint DA (2002) Trek-like potassium channels in rat cardiac ventricular myocytes are activated by intracellular ATP. J Membr Biol 185:201–207

    Article  PubMed  CAS  Google Scholar 

  36. Tavernarakis N, Driscoll M (1997) Molecular modelling of mechanotransduction in the nematode Caenorhabditis elegans. Annu Rev Physiol 59:659–689

    Article  PubMed  CAS  Google Scholar 

  37. Ullrich ND, Voets T, Prenen J, Vennekens R, Talavera K, Droogmans G, Nilius B (2005) Comparison of functional properties of the Ca2+-activated cation channels TRPM4 and TRPM5 from mice. Cell Calcium 37:267–278

    Article  PubMed  CAS  Google Scholar 

  38. Vriens J, Watanabe H, Janssens A, Droogmans G, Voets T, Nilius B (2004) Cell swelling, heat, and chemical agonists use distinct pathways for the activation of the cation channel TRPV4. Proc Natl Acad Sci U S A 101:396–401

    Article  PubMed  CAS  Google Scholar 

  39. Walker RG, Willingham AT, Zuker CS (2000) A Drosophila mechanosensory transduction channel. Science 287:2229–2234

    Article  PubMed  CAS  Google Scholar 

  40. Wang GX, Poo M (2005) Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones. Nature 434:898–904

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

The authors wish to thank Paolo Orsini who developed part of the calcium imaging software and Francesco Montanari for making the filter wheel device.

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Authors and Affiliations

  1. Dipartimento di Fisiologia e Biochimica “G. Moruzzi”, Università di Pisa, Via S. Zeno 31, 56127,, Pisa, Italy

    C. Barsanti, D. Ricci & M. Pellegrino

  2. Scuola Normale Superiore, piazza Cavalieri 7, 56126,, Pisa, Italy

    M. Pellegrini

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  1. C. Barsanti
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  2. M. Pellegrini
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  3. D. Ricci
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  4. M. Pellegrino
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Correspondence to M. Pellegrino.

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Barsanti, C., Pellegrini, M., Ricci, D. et al. Effects of intracellular pH and Ca2+ on the activity of stretch-sensitive cation channels in leech neurons. Pflugers Arch - Eur J Physiol 452, 435–443 (2006). https://doi.org/10.1007/s00424-006-0056-7

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  • DOI: https://doi.org/10.1007/s00424-006-0056-7

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