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Pain Research pp 73–97Cite as

Whole-Cell Recording in Isolated Primary Sensory Neurons

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 851))

Abstract

The isolated sensory neuron in vitro is a powerful model with which to address a number of important neurobiological questions. Isolated neurons are relatively easy to prepare from both neonatal and adult animals and can be studied both acutely and after considerable time on culture. Intracellular recording is one of the most powerful ways to study these neurons. Methods are described for both the preparation of isolated sensory neurons in vitro as well as for recording major classes of ionic currents (Na+, K+, and Ca2+) from these neurons with whole cell voltage-clamp techniques. Methods are also provided for an initial characterization of active and passive electrophysiological properties of these neurons in current clamp as well as the use of perforated patch recording as a means to mitigate some of the limitations associated with conventional whole cell patch recording. The reader should be aware that the regulation of ion channels in sensory neurons may very subtle, requiring considerably more sophisticated protocols than have been provided here. The reader should also be aware that there is a tremendous heterogeneity among sensory neurons, which is both a curse and a blessing for those who wish to study them. Thus, the methods provided here should only be considered the starting point for a more detailed analysis of sensory neurons.

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References

  1. McCarter, G. C., Reichling, D. B., and Levine, J. D. (1999) Mechanical transduction by rat dorsal root ganglion neurons in vitro, Neurosci Lett 273, 179–182.

    Article  PubMed  CAS  Google Scholar 

  2. Caterina, M. J., Schumacher, M. A., Tominaga, M., Rosen, T. A., Levine, J. D., and Julius, D. (1997) The capsaicin receptor: a heat-activated ion channel in the pain pathway, Nature 389, 816–824.

    Article  PubMed  CAS  Google Scholar 

  3. Thut, P. D., Wrigley, D., and Gold, M. S. (2003) Cold transduction in rat trigeminal ganglia neurons in vitro, Neuroscience 119, 1071–1083.

    Article  PubMed  CAS  Google Scholar 

  4. Gold, M. S., and Caterina, M. J. (2008) Molecular Biology of Nociceptor Transduction, in The Senses: A Comprehensive Reference (Basbaum, A. I., and Bushnell, M. C., Eds.), pp 43–74, Academic Press, San Diego.

    Chapter  Google Scholar 

  5. Flake, N. M., Lancaster, E., Weinreich, D., and Gold, M. S. (2004) Absence of an association between axotomy-induced changes in sodium currents and excitability in DRG neurons from the adult rat, Pain 109, 471–480.

    Article  PubMed  CAS  Google Scholar 

  6. Flake, N. M., Bonebreak, D. B., and Gold, M. S. (2005) Estrogen and inflammation increase the excitability of rat temporomandibular joint afferent neurons, J Neurophysiol 93, 1585–1597.

    Article  PubMed  CAS  Google Scholar 

  7. Harriott, A. M., Dessem, D., and Gold, M. S. (2006) Inflammation increases the excitability of masseter muscle afferents, Neuroscience 141, 433–442.

    Article  PubMed  CAS  Google Scholar 

  8. Huang, L. Y., and Neher, E. (1996) Ca(2+)-dependent exocytosis in the somata of dorsal root ganglion neurons, Neuron 17, 135–145.

    Article  PubMed  CAS  Google Scholar 

  9. Tomkins, C. E., Edwards, S. N., and Tolkovsky, A. M. (1994) Apoptosis is induced in post-mitotic rat sympathetic neurons by arabinosides and topoisomerase II inhibitors in the presence of NGF, J Cell Sci 107, 1499–1507.

    PubMed  CAS  Google Scholar 

  10. Hamill, O. P., Marty, A., Neher, E., Sakmann, B., and Sigworth, F. J. (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 

  11. Sakmann, B., and Neher, E., (Eds.) (1995) Single-Channel Recording, Second ed., Plenum, New York.

    Google Scholar 

  12. Ho, C., and O’Leary, M. E. (2011) Single-cell analysis of sodium channel expression in dorsal root ganglion neurons, Mol Cell Neurosci. 46(1), 159–166.

    Google Scholar 

  13. Waxman, S. G., Kocsis, J. D., and Black, J. A. (1994) Type III sodium channel mRNA is expressed in embryonic but not adult spinal sensory neurons, and is reexpressed following axotomy, J Neurophysiol 72, 466–470.

    PubMed  CAS  Google Scholar 

  14. Renganathan, M., Dib-Hajj, S., and Waxman, S. G. (2002) Na(v)1.5 underlies the ‘third TTX-R sodium current’ in rat small DRG neurons, Brain Res Mol Brain Res 106, 70–82.

    Article  PubMed  CAS  Google Scholar 

  15. Xiao, Y., Blumenthal, K., Jackson, J. O., 2nd, Liang, S., and Cummins, T. R. The tarantula toxins ProTx-II and huwentoxin-IV differentially interact with human Nav1.7 voltage sensors to inhibit channel activation and inactivation, Mol Pharmacol 78, 1124–1134.

    Google Scholar 

  16. Coste, B., Osorio, N., Padilla, F., Crest, M., and Delmas, P. (2004) Gating and modulation of presumptive NaV1.9 channels in enteric and spinal sensory neurons, Mol Cell Neurosci 26, 123–134.

    Article  PubMed  CAS  Google Scholar 

  17. Cummins, T. R., Dib-Hajj, S. D., Black, J. A., Akopian, A. N., Wood, J. N., and Waxman, S. G. (1999) A novel persistent tetrodotoxin-resistant sodium current in SNS-null and wild-type small primary sensory neurons, J Neurosci 19, RC43.

    Google Scholar 

  18. Gold, M. S., Shuster, M. J., and Levine, J. D. (1996) Characterization of six voltage-gated K+ currents in adult rat sensory neurons, J. Neurophysiol. 75, 2629–2646.

    PubMed  CAS  Google Scholar 

  19. Zhang, X. L., Mok, L. P., Katz, E. J., and Gold, M. S. BKCa currents are enriched in a subpopulation of adult rat cutaneous nociceptive dorsal root ganglion neurons, Eur J Neurosci 31, 450–462.

    Google Scholar 

  20. Lu, S. G., Zhang, X. L., Luo, Z. D., and Gold, M. S. Persistent inflammation alters the density and distribution of voltage-activated calcium channels in subpopulations of rat cutaneous DRG neurons, Pain 151, 633–643.

    Google Scholar 

  21. Harriott, A. M., and Gold, M. S. (2009) Electrophysiological Properties of Dural Afferents in the Absence and Presence of Inflammatory Mediators, J Neurophysiol. 101(6), 3126–3134.

    Google Scholar 

  22. Magistretti, J., Mantegazza, M., Guatteo, E., and Wanke, E. (1996) Action potentials recorded with patch-clamp amplifiers: are they genuine? Trends Neurosci 19, 530–534.

    Article  PubMed  CAS  Google Scholar 

  23. Nicol, G. D., Vasko, M. R., and Evans, A. R. (1997) Prostaglandins suppress an outward potassium current in embryonic rat sensory neurons, J Neurophysiol 77, 167–176.

    PubMed  CAS  Google Scholar 

  24. Gold, M. S., Shuster, M. J., and Levine, J. D. (1996) Role of a Ca(2+)-dependent slow afterhyperpolarization in prostaglandin E2-induced sensitization of cultured rat sensory neurons, Neurosci Lett 205, 161–164.

    Article  PubMed  CAS  Google Scholar 

  25. Scroggs, R. S., and Fox, A. P. (1992) Calcium current variation between acutely isolated adult rat dorsal root ganglion neurons of different size, Journal of Physiology 445, 639–658.

    PubMed  CAS  Google Scholar 

  26. Scroggs, R. S., Todorovic, S. M., Anderson, E. G., and Fox, A. P. (1994) Variation in IH, IIR, and ILEAK between acutely isolated adult rat dorsal root ganglion neurons of different size, J Neurophysiol 71, 271–279.

    CAS  Google Scholar 

  27. Cardenas, C. G., Del Mar, L. P., and Scroggs, R. S. (1995) Variation in serotonergic inhibition of calcium channel currents in four types of rat sensory neurons differentiated by membrane properties, J Neurophysiol 74, 1870–1879.

    PubMed  CAS  Google Scholar 

  28. Eckert, S. P., Taddese, A., and McCleskey, E. W. (1998) Isolation and culture of rat sensory neurons having distinct sensory modalities, J Neurosci Meth 77, 183–190.

    Article  Google Scholar 

  29. White, G., Lovinger, D. M., and Weight, F. F. (1989) Transient low-threshold Ca2+ current triggers burst firing through an afterdepolarizing potential in an adult mammalian neuron, Proceedings of the National Academy of Sciences 86, 6802–6806.

    Article  CAS  Google Scholar 

  30. Delree, P., Ribbens, C., Martin, D., Rogister, B., Lefebvre, P. P., Rigo, J. M., Leprince, P., Schoenen, J., and Moonen, G. (1993) Plasticity of developing and adult dorsal root ganglion neurons as revealed in vitro, Brain Research Bulletin 30, 231–237.

    Article  PubMed  CAS  Google Scholar 

  31. Winter, J. (1998) Brain derived neurotrophic factor, but not nerve growth factor, regulates capsaicin sensitivity of rat vagal ganglion neurones, Neurosci Lett 241, 21–24.

    Article  PubMed  CAS  Google Scholar 

  32. Akaike, N. (1996) Gramicidin perforated patch recording and intracellular chloride activity in excitable cells, Prog Biophys Mol Biol 65, 251–264.

    Article  PubMed  CAS  Google Scholar 

  33. Vaughn, A. H., and Gold, M. S. Ionic mechanisms underlying inflammatory mediator-induced sensitization of dural afferents, J Neurosci 30, 7878–7888.

    Google Scholar 

  34. Price, T. J., Cervero, F., Gold, M. S., Hammond, D. L., and Prescott, S. A. (2009) Chloride regulation in the pain pathway, Brain Res Rev 60, 149–170.

    Article  PubMed  CAS  Google Scholar 

  35. Isom, L. L. (2001) Sodium channel beta subunits: anything but auxiliary, Neuroscientist 7, 42–54.

    Article  PubMed  CAS  Google Scholar 

  36. Fitzgerald, E. M., Okuse, K., Wood, J. N., Dolphin, A. C., and Moss, S. J. (1999) Cyclic AMP-dependent phosphorylation of the tetrodotoxin-resistant voltage-dependent sodium channel, SNS, J Physiol. 516(Pt 2), 433–446.

    Google Scholar 

  37. Reimann, F., Cox, J. J., Belfer, I., Diatchenko, L., Zaykin, D. V., McHale, D. P., Drenth, J. P., Dai, F., Wheeler, J., Sanders, F., Wood, L., Wu, T. X., Karppinen, J., Nikolajsen, L., Mannikko, M., Max, M. B., Kiselycznyk, C., Poddar, M., Te Morsche, R. H., Smith, S., Gibson, D., Kelempisioti, A., Maixner, W., Gribble, F. M., and Woods, C. G. Pain perception is altered by a nucleotide polymorphism in SCN9A, Proc Natl Acad Sci U S A 107, 5148–5153.

    Google Scholar 

  38. Chien, L. Y., Cheng, J. K., Chu, D., Cheng, C. F., and Tsaur, M. L. (2007) Reduced expression of A-type potassium channels in primary sensory neurons induces mechanical hypersensitivity, J Neurosci 27, 9855–9865.

    Article  PubMed  CAS  Google Scholar 

  39. Gold, M. S., Dastmalchi, S., and Levine, J. D. (1996) Co-expression of nociceptor properties in dorsal root ganglion neurons from the adult rat in vitro, Neuroscience 71, 265–275.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by NIH grants DE018252 and NS063010. I would like to thank Nicole Scheff for valuable input on the preparation of this manuscript.

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Authors and Affiliations

  1. Department of Anesthesiology, University of Pittsburgh, Pittsburgh, PA, USA

    Michael S. Gold

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  1. Michael S. Gold
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Correspondence to Michael S. Gold .

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Editors and Affiliations

  1. Irvine Medical Center, Dept. Anesthesiology & Pharmacology, University of California, The City Drive S. 101, Orange, 92868, California, USA

    Z. David Luo

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Gold, M.S. (2012). Whole-Cell Recording in Isolated Primary Sensory Neurons. In: Luo, Z. (eds) Pain Research. Methods in Molecular Biology, vol 851. Humana Press. https://doi.org/10.1007/978-1-61779-561-9_5

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  • DOI: https://doi.org/10.1007/978-1-61779-561-9_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-560-2

  • Online ISBN: 978-1-61779-561-9

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