Skip to main content
SpringerLink
Log in

Whole-Cell Patch Clamp Recordings from Oligodendrocyte Lineage Cells in Brain Slices

  • Protocol
  • First Online:
Oligodendrocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1936))

Abstract

The whole-cell configuration of the patch-clamp technique is widely used to study electrically active cells and passive membrane properties, as well as the properties and pharmacology of ion channels, neurotransmitter receptors, and electrogenic transporters, in almost any cell type. In the brain, in addition to neurons, oligodendrocyte precursor cells (OPCs) that give rise to myelinating oligodendrocytes (OLs) are also excitable. Electrophysiological techniques provide the main tool for the thorough investigation of the electrogenic capacity of such cell types. Although there are many published protocols for whole-cell recordings, there are very few that touch upon the electrophysiological characteristics of oligodendrocyte lineage cells. Here we provide a detailed methodology for how to acquire and analyze whole-cell recordings from excitable cells, with a focus on oligodendrocyte lineage cells. We provide a protocol on how to successfully identify OPCs and OLs in brain slices, either with the use of transgenic animal models or through morphological and electrophysiological profiling. The method described can also be easily adopted for whole-cell recordings from oligodendrocyte lineage cells in vitro

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 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
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. Neher E, Sakmann B (1976) Single-channel currents recorded from membrane of denervated frog muscle fibres. Nature 260(5554):799–802

    Article  CAS  Google Scholar 

  2. 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(2):85–100

    Article  CAS  Google Scholar 

  3. Sakmann B, Neher E (1984) Patch clamp techniques for studying ionic channels in excitable membranes. Annu Rev Physiol 46(1):455–472

    Article  CAS  Google Scholar 

  4. Hodgkin AL, Huxley AF (1952) Currents carried by sodium and potassium ions through the membrane of the giant axon of Loligo. J Physiol 116(4):449–472

    Article  CAS  Google Scholar 

  5. Bergles DE, Roberts JD, Somogyi P, Jahr CE (2000) Glutamatergic synapses on oligodendrocyte precursor cells in the hippocampus. Nature 405(6783):187–191

    Article  CAS  Google Scholar 

  6. Chittajallu R, Aguirre A, Gallo V (2004) NG2-positive cells in the mouse white and grey matter display distinct physiological properties. J Physiol 561(Pt 1):109–122

    Article  CAS  Google Scholar 

  7. Káradóttir R, Cavelier P, Bergersen LH, Attwell D (2005) NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. Nature 438(7071):1162–1166

    Article  Google Scholar 

  8. Bakiri Y, Burzomato V, Frugier G, Hamilton NB, Káradóttir R, Attwell D (2009) Glutamatergic signaling in the brain’s white matter. Neuroscience 158(1):266–274

    Article  CAS  Google Scholar 

  9. Káradóttir R, Hamilton NB, Bakiri Y, Attwell D (2008) Spiking and nonspiking classes of oligodendrocyte precursor glia in CNS white matter. Nat Neurosci 11(4):450–456

    Article  Google Scholar 

  10. Spitzer S, Volbracht K, Lundgaard I, Káradóttir RT (2016) Glutamate signalling: a multifaceted modulator of oligodendrocyte lineage cells in health and disease. Neuropharmacology 110:574–585

    Article  CAS  Google Scholar 

  11. Káradóttir R, Attwell D (2007) Neurotransmitter receptors in the life and death of oligodendrocytes. Neuroscience 145(4):1426–1438

    Article  Google Scholar 

  12. Marinelli C, Bertalot T, Zusso M, Skaper SD, Giusti P (2016) Systematic review of pharmacological properties of the oligodendrocyte lineage. Front Cell Neurosci 10:27

    Article  Google Scholar 

  13. Kukley M, Capetillo-Zarate E, Dietrich D (2007) Vesicular glutamate release from axons in white matter. Nat Neurosci 10(3):311–320

    Article  CAS  Google Scholar 

  14. Ziskin JL, Nishiyama A, Rubio M, Fukaya M, Bergles DE (2007) Vesicular release of glutamate from unmyelinated axons in white matter. Nat Neurosci 10(3):321–330

    Article  CAS  Google Scholar 

  15. Gautier HOB, Evans KA, Volbracht K, James R, Sitnikov S, Lundgaard I et al (2015) Neuronal activity regulates remyelination via glutamate signalling to oligodendrocyte progenitors. Nat Commun 6:8518

    Article  CAS  Google Scholar 

  16. Young KM, Psachoulia K, Tripathi RB, Dunn S-J, Cossell L, Attwell D et al (2013) Oligodendrocyte dynamics in the healthy adult CNS: evidence for myelin remodeling. Neuron 77(5):873–885

    Article  CAS  Google Scholar 

  17. McKenzie IA, Ohayon D, Li H, Paes de Faria J, Emery B, Tohyama K et al (2014) Motor skill learning requires active central myelination. Science 346(6207):318–322

    Article  CAS  Google Scholar 

  18. Lundgaard I, Luzhynskaya A, Stockley JH, Wang Z, Evans KA, Swire M et al (2013) Neuregulin and BDNF induce a switch to NMDA receptor-dependent myelination by oligodendrocytes. PLoS Biol 11(12):e1001743

    Article  Google Scholar 

  19. Gibson EM, Purger D, Mount CW, Goldstein AK, Lin GL, Wood LS et al (2014) Neuronal activity promotes oligodendrogenesis and adaptive myelination in the mammalian brain. Science 344(6183):1252304

    Article  Google Scholar 

  20. Verkhratsky A, Parpura V et al (2014) Methods Mol Biol 1183:1–19

    Article  Google Scholar 

  21. Sontheimer H (1995) Whole-cell patch-clamp recordings, in: Patch-clamp applications and protocols. Humana Press, New Jersey, pp. 37–74. https://doi.org/10.1385/0-89603-311-2:37

  22. Gibb JA, Edwards FA (1994). Patch clamp recording from cells in sliced tissues. In: Ogden D (Second Eds.) Plymouth workshop. Microelectrode techniques: the plymouth workshop handbook. Company of Biologists, pp. 255–274

    Google Scholar 

  23. Moyer JR, Brown TH (1966) Patch clamping chapter 5. Dent Tech 35:135–193

    Google Scholar 

  24. Molleman A (2003) Patch clamping: an introductory guide to patch clamp electrophysiology, vol 186. Wiley, Chichester

    Google Scholar 

  25. Kornreich BG (2007) The patch clamp technique: principles and technical considerations. J Vet Cardiol 9(1):25–37

    Article  Google Scholar 

  26. Sakmann B, Neher E (1995) Single-channel recording. Springer, New York, p 700

    Google Scholar 

  27. Karram K, Goebbels S, Schwab M, Jennisen K, Seifert G, Steinhäuser C et al (2008) NG2-expressing cells in the nervous system revealed by the NG2-EYFP-knock in mouse. Genesis 46(12):743–757

    Article  CAS  Google Scholar 

  28. Chen Y, Spitzer S, Agathou S, Karadottir RT, Smith A (2017) Gene editing in rat embryonic stem cells to produce in vitro models and in vivo reporters. Stem Cell Reports 9:1262–1274

    Article  CAS  Google Scholar 

  29. Blanco S, Dietmann S, Flores JV, Hussain S, Kutter C, Humphreys P et al (2014) Aberrant methylation of tRNAs links cellular stress to neuro-developmental disorders. EMBO J 33(18):1–20

    Article  Google Scholar 

  30. Spitzer S, Sitnikov S, Kamen Y, Evans KA, Kronenberg-Versteeg D, Dietmann S, de Faria O, Agathou S, Káradóttir R (2018) Oligodendrocyte progenitor cells become regionally diverse and heterogeneous with age, Neuron. (in press)

    Google Scholar 

  31. Káradóttir R, Attwell D (2006) Combining patch-clamping of cells in brain slices with immunocytochemical labeling to define cell type and developmental stage. Nat Protoc 1(4):1977–1986

    Article  Google Scholar 

  32. Tessier-Lavigne M, Attwell D, Mobbs P, Wilson M (1988) Membrane currents in retinal bipolar cells of the axolotl. J Gen Physiol 91(1):49–72

    Article  CAS  Google Scholar 

  33. Major G (1993) Solutions for transients in arbitraruly branching cables: III voltage clamp problems. Biophys J 65(1):469–491

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Veterinary Medicine, Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK

    Sylvia Agathou & Ragnhildur Thóra Káradóttir

Authors
  1. Sylvia Agathou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ragnhildur Thóra Káradóttir
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ragnhildur Thóra Káradóttir .

Editor information

Editors and Affiliations

  1. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    David A. Lyons

  2. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    Linde Kegel

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Agathou, S., Káradóttir, R.T. (2019). Whole-Cell Patch Clamp Recordings from Oligodendrocyte Lineage Cells in Brain Slices. In: Lyons, D., Kegel, L. (eds) Oligodendrocytes. Methods in Molecular Biology, vol 1936. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9072-6_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9072-6_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9070-2

  • Online ISBN: 978-1-4939-9072-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation