Skip to main content
SpringerLink
Log in

Patch-Cram Detection of Cyclic GMP in Intact Cells

  • Protocol
Patch-Clamp Analysis

Part of the book series: Neuromethods ((NM,volume 35))

  • 1100 Accesses

Abstract

Information is transmitted throughout the nervous system by chemical and electrical signals. The cacophony of neuronal signals does not end at the cell surface, but continues deep into cells via diffusible intracellular messenger molecules. The list of molecules that serve as intracellular messengers continues to expand, but several stand out as very common examples, including Ca2+, cyclic nucleotides (cAMP and cGMP), and inositol phospholipids and their metabolites (IP3, diacylglycerol, etc.). Cellular responses to extracellular neurotransmitters are often mediated by one or more of these intracellular messengers, affording several functional advantages. Intracellular messengers allow spatial spread of signals within cells, contribute to amplification of signals, and allow responses to persist even after the neurotransmitter is gone from outside the cell. Intracellular messenger systems are also advantageous because they participate in a great degree of “crosstalk,” increasing the repertoire of responses to a given external stimulus and allowing for biochemical computation at the level of an individual cell.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  • Adams, S. R., Harootunian, A. T., Buechler, Y. J., et al. (1991) Fluorescence ratio imaging of cyclic AMP in single cells. Nature 349, 694–697.

    Article  PubMed  CAS  Google Scholar 

  • Ando, M., Tatematsu, T., Kunii, S., and Nagata, Y. (1994) The intercellular communication via nitric oxide and its regulation in coupling of cyclic GMP synthesis upon stimulation of muscarinic cholinergic receptors in rat superior cervical sympathetic ganglia. Brain Res. 650, 283–288.

    Article  PubMed  CAS  Google Scholar 

  • Backsaw, B. J., Hochner, B., Mahaut-Smith, M., Adams, S. R., Kaang, B. K., Kandel, E. R., and Tsien, R. Y. (1993) Spatially resolved dynamics of cAMP and protein kinase A subunits in Aplysia sensory neurons. Science 260, 222–226.

    Article  Google Scholar 

  • Barria, A., Muller, D., Derkach, V., Griffith, L. C., and Soderling, T. R. (1997) Regulatory phophorylation of AMPA-type glutamate receptors by CaM KII during long-term potentiation. Science 276, 2042–2045.

    Article  PubMed  CAS  Google Scholar 

  • Briggs, C. A., Horwitz, J., McAfee, D. A., Tsymbalov, S., and Perlman, R. L. (1985) Effects of neuronal activity on inositol phospholipid metabolism in the rat autonomic nervous system. J. Neurochem. 44, 731–739.

    Article  PubMed  CAS  Google Scholar 

  • Broillet, M. C. and Firestein, S. (1996) Direct activation of the olfactory cyclic nucleotide-gated channel through modification of sulfhydryl groups by NO compounds. Neuron 16, 377–385.

    Article  PubMed  CAS  Google Scholar 

  • Catarsi, S. and Drapeau, P. (1996) Modulation and selection of neurotransmitter responses during synapse formation between identified leech neurons. Cell. Mol. Neurobiol. 16, 699–713.

    Article  PubMed  CAS  Google Scholar 

  • Civitelli, R., Bacskai, B. J., Mahaut-Smith, M. P., Adams, S. R., Avioli, L. V., and Tsien, R. Y. (1994) Single-cell analysis of cyclic AMP response to parathyroid hormone in osteoblastic cells. J. Bone Miner. Res. 9, 1407–1417.

    Article  PubMed  CAS  Google Scholar 

  • Costantin, J. L., Qin, N., Waxham, M. N., Birnbaumer, L., and Stefani, E. (1999) Complete reversal of run-down in rabbit cardiac Ca2+channels by patchcramming in Xenopus oocytes; partial reversal by protein kinase A. Pflugers Arch. 437, 888–894.

    Article  PubMed  CAS  Google Scholar 

  • Dhallan, R. S., Yau, K. W., Schrader, K. A., and Reed, R. R. (1990) Primary structure and functional expression of a cyclic nucleotide-activated channel from olfactory neurons. Nature 347, 184–187.

    Article  PubMed  CAS  Google Scholar 

  • Horn, R. and Marty, A. (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J. Gen. Physiol. 92, 145–159.

    Article  PubMed  CAS  Google Scholar 

  • Forstermann, U., Gorsky, L. D., Pollock, J. S., et al. (1990) Hormone-induced biosynthesis of endothelium-derived relaxing factor/nitric oxide-like material in N1E-115 neuroblastoma cells requires calcium and calmodulin. Mol. Pharmacol. 38, 7–13.

    PubMed  CAS  Google Scholar 

  • Garbers, D. L. (1981) Guanylyl cyclase-linked receptors. Pharmacol. Ther. 50, 337–345.

    Article  Google Scholar 

  • Garthwaite, J. (1991) Glutamate, nitric oxide and cell-cell signalling in the nervous system. Trends Neurosci. 14, 60–67.

    Article  PubMed  CAS  Google Scholar 

  • Garthwaite, J., Southam, E., Boulton, C. L., Nielsen, E. B., Schmidt, K., and Mayer, B. (1995) Potent and selective inhibition of nitric oxide-sensitive guanylyl cyclaseby 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. Mol. Pharmacol. 48, 184–188.

    PubMed  CAS  Google Scholar 

  • Goulding, E. H., Tibbs, G. R., and Siegelbaum, S. A. (1994) Molecular mechanism of cyclic-nucleotide-gated channel activation. Nature 372, 369–374.

    Article  PubMed  CAS  Google Scholar 

  • Hempel, C. M., Vincent, P., Adams, S. R., Tsien, R. Y., and Selverston, A. I. (1996) Spatio-temporal dynamics of cyclic AMP signals in an intact neural circuitm. Nature 384, 166–169.

    Article  PubMed  CAS  Google Scholar 

  • Hobbs, A. J. (1997) Soluble guanylate cyclase: the forgotten sibling. Trends Pharmacol. Sci. 18, 484–491.

    PubMed  CAS  Google Scholar 

  • Horn, R. and Marty, A. (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J. Gen. Physiol. 92, 145–159.

    Article  PubMed  CAS  Google Scholar 

  • Honda, A., Adams, S. R., Sawyer, C. L., Lev-Ram, V. V., Tsien, R. Y., and Dostmann, W. R. (2001) Spatiotemporal dynamics of guanosine 3′,5′-cyclic monophosphate revealed by a genetically encoded, fluorescent indicator. Proc. Natl. Acad. Sci. USA 98, 2437–2442.

    Article  PubMed  CAS  Google Scholar 

  • Horn, R. and Korn, S. J. (1992) Prevention of rundown in electrophysiological recording. Methods Enzymol. 207, 149–155.

    Article  PubMed  CAS  Google Scholar 

  • Hu, J. and el-Fakahany, E. E. (1993) Role of intercellular and intracellular communication by nitric oxide in coupling of muscarinic receptors to activation of guanylate cyclase in neuronal cells. J. Neurochem. 61, 578–585.

    Article  PubMed  CAS  Google Scholar 

  • Kimhi, Y., Palfrey, C., Spector, I., Barak, Y., and Littauer, U. Z. (1976) Maturation of neuroblastoma cells in the presence of dimethylsulfoxide. Proc. Natl. Acad. Sci. USA 73, 462–466.

    Article  PubMed  CAS  Google Scholar 

  • Kramer, R. H. (1990) Patch cramming: monitoring intracellular messengers in intact cells with membrane patches containing detector ion channels. Neuron 4, 335–341.

    Article  PubMed  CAS  Google Scholar 

  • Lev-Ram, V., Jiang, T., Wood, J., Lawrence, D. S., and Tsein, R. Y. (1997) Synergies and coincidence requirements between NO, cGMP, and Ca2+in the induction of cerebellar long-term depression. Neuron 18, 1025–1038.

    Article  PubMed  CAS  Google Scholar 

  • Malinow, R., Madison, D. V., and Tsein, R. W. (1988) Persistent protein kinase activity underlying long-term potentiation. Nature 335, 820–824.

    Article  PubMed  CAS  Google Scholar 

  • Mathes C. and Thompson, S. H. (1996) The nitric oxide/cGMP pathway couples muscarinic receptors to the activation of Ca2+influx. J. Neurosci. 16, 1702–1709.

    PubMed  CAS  Google Scholar 

  • Molokanova, E., Maddox, F., Luetje, C. W., and Kramer, R. H. (1999) Activitydependent modulation of rod photoreceptor cyclic nucleotide-gated channels mediated by phosphorylation of a specific tyrosine residue. J. Neurosci. 19, 4786–4795.

    PubMed  CAS  Google Scholar 

  • Molokanova, E., Trivedi, B., Savchenko, A., and Kramer, R. H. (1997) Modulation of rod photoreceptor cyclic nucleotide-gated channels by tyrosine phosphorylation. J. Neurosci. 17, 9068–9076.

    PubMed  CAS  Google Scholar 

  • Moncada, S. and Higgs, E. A. (1995) Molecular mechanisms and therapeutic strategies related to nitric oxide. FASEB J. 9, 1319–1330.

    PubMed  CAS  Google Scholar 

  • Tang, X. D. and Hoshi, T. (1999) Rundown of the hyperpolarization-activated KAT1 channel involves slowing of the opening transitions regulated by phosphorylation. Biophys. J. 76, 3089–3098.

    Article  PubMed  CAS  Google Scholar 

  • Thompson, S. H., Mathes, C., and Alousi, A. A. (1995) Calcium requirement for cGMP production during muscarinic activation of N1E-115 neuroblastoma cells. Am. J. Physiol. 269, C979–C985.

    PubMed  CAS  Google Scholar 

  • Wotta, D. R., Parsons, A. M., Hu, J., Grande, A. W., and El-Fakahany, E. E. (1998) M1 muscarninic receptors stimulate rapid and prolonged phases of neuronal nitric oxide synthase activity: involvement of different calcium pools. J. Neurochem. 71, 487–497.

    Article  PubMed  CAS  Google Scholar 

  • Zaccolo, M., De Giorgi, F., Cho, C. Y., Feng, L., Knapp, T., Negulescu, P. A., et al. (2000) A genetically encoded, fluorescent indicator for cyclic AMP in living cells. Nat. Cell Biol. 2, 25–29.

    Article  PubMed  CAS  Google Scholar 

  • Zagotta, W. N. and Siegelbaum, S. A. (1996) Structure and function of cyclic nucleotide-gated channels. Annu. Rev. Neurosci. 19, 235–263.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California, Berkeley, CA

    Richard H. Kramer

Authors
  1. Richard H. Kramer
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Saskatchewan, Saskatoon, Canada

    Alan A. Boulton , Glen B. Baker  & Wolfgang Walz ,  & 

  2. Alviva Biopharmaceuticals Inc., Saskatoon, Canada

    Alan A. Boulton

  3. University of Alberta, Edmonton, Canada

    Glen B. Baker

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Humana Press Inc.

About this protocol

Cite this protocol

Kramer, R.H. (2002). Patch-Cram Detection of Cyclic GMP in Intact Cells. In: Walz, W., Boulton, A.A., Baker, G.B. (eds) Patch-Clamp Analysis. Neuromethods, vol 35. Humana Press. https://doi.org/10.1385/1-59259-276-7:245

Download citation

  • DOI: https://doi.org/10.1385/1-59259-276-7:245

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-013-7

  • Online ISBN: 978-1-59259-276-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation