Skip to main content

Outside-Out “Sniffer-Patch” Clamp Technique for In Situ Measures of Neurotransmitter Release

  • Protocol
  • First Online:
Patch-Clamp Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1183))

Abstract

The mechanism underlying neurotransmitter release is a critical research domain for the understanding of neuronal network function; however, few techniques are available for the direct detection and measurement of neurotransmitter release. To date, the sniffer-patch clamp technique is mainly used to investigate these mechanisms from individual cultured cells. In this study, we propose to adapt the sniffer-patch clamp technique to in situ detection of neurosecretion. Using outside-out patches from donor cells as specific biosensors plunged in acute cerebral slices, this technique allows for proper detection and quantification of neurotransmitter release at the level of the neuronal network.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Horn R, Patlak J (1980) Single channel currents from excised patches of muscle membrane. Proc Natl Acad Sci U S A 77:6930–6934

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Young SH, Poo MM (1992) Measurement of transmitter release: moving patch method. In: Kettenmann H, Grantyn R (eds) Practical electrophysiological methods. Wisley-Liss, New York, pp 354–357

    Google Scholar 

  3. Hume RI, Role LW, Fischbach GD (1983) Acetylcholine release from growth cones detected with patches of acetylcholine receptor-rich membranes. Nature 305:632–634

    Article  CAS  PubMed  Google Scholar 

  4. Young SH, Poo MM (1983) Spontaneous release of transmitter from growth cones of embryonic neurones. Nature 305:634–637

    Article  CAS  PubMed  Google Scholar 

  5. Grinnell AD, Gundersen CB, Meriney SD et al (1989) Direct measurement of Ach release from exposed frog nerve terminals: constraints on interpretation of non-quantal release. J Physiol 419:225–251

    CAS  PubMed Central  PubMed  Google Scholar 

  6. Silinsky EM, Redman RS (1996) Synchronous release of ATP and neurotransmitter within milliseconds of a motor nerve impulse in the frog. J Physiol 492:815–822

    CAS  PubMed Central  PubMed  Google Scholar 

  7. Maeda T, Shimoshige Y, Mizukami K et al (1995) Patch sensor detection of glutamate release evoked by a single electrical shock. Neuron 15:253–257

    Article  CAS  PubMed  Google Scholar 

  8. Billups B, Attwell D (1996) Modulation of non-vesicular glutamate release by pH. Nature 379:171–174

    Article  CAS  PubMed  Google Scholar 

  9. Shimane M, Miyagawa K, Sugawara M (2006) Detection of gamma-aminobutyric acid-induced glutamate release in acute mouse hippocampal slices with a patch sensor. Anal Biochem 353:83–92

    Article  CAS  PubMed  Google Scholar 

  10. Scain AL, Le Corronc H, Allain AE et al (2010) Glycine release from radial cells modulates the spontaneous activity and its propagation during early spinal cord development. J Neurosci 30:390–403

    Article  CAS  PubMed  Google Scholar 

  11. Mangin JM, Baloul M, Prado De Carvalho L et al (2003) Kinetic properties of the alpha2 homo-oligomeric glycine receptor impairs a proper synaptic functioning. J Physiol 553:369–586

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  13. Van-Hooft JA, Vijverberg HP (1995) Phosphorylation controls conductance of 5-HT3 receptor ligand-gated ion channels. Receptors Channels 3:7–12

    CAS  PubMed  Google Scholar 

  14. Martina M, Krasteniakov NV, Bergeron R (2003) D-Serine differently modulates NMDA receptor function in rat CA1 hippocampal pyramidal cells and interneurons. J Physiol 548:411–423

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Martina M, Gorfinkel Y, Halman S et al (2004) Glycine transporter type 1 blockade changes NMDA receptor-mediated responses and LTP in hippocampal CA1 pyramidal cells by altering extracellular glycine levels. J Physiol 557: 489–500

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Martina M, Turcotte ME, Halman S et al (2007) The sigma-1 receptor modulates NMDA receptor synaptic transmission and plasticity via SK channels in rat hippocampus. J Physiol 578:143–157

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Muller E, Bakkar W, Martina M et al (2013) Vesicular storage of glycine in glutamatergic terminals in mouse hippocampus. Neuroscience 242:110–127

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Émilie Muller-Chrétien .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Muller-Chrétien, É. (2014). Outside-Out “Sniffer-Patch” Clamp Technique for In Situ Measures of Neurotransmitter Release. In: Martina, M., Taverna, S. (eds) Patch-Clamp Methods and Protocols. Methods in Molecular Biology, vol 1183. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1096-0_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1096-0_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1095-3

  • Online ISBN: 978-1-4939-1096-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics