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Monitoring local synaptic activity with astrocytic patch pipettes

Nature Protocols volume 7, pages 2171–2179 (2012)Cite this article

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Abstract

Rapid signal exchange between astroglia and neurons has emerged as a key player in neural communication in the brain. To understand the mechanisms involved, it is often important to have access to individual astrocytes while monitoring the activity of nearby synapses. Achieving this with standard electrophysiological tools is not always feasible. The protocol presented here enables the monitoring of synaptic activity using whole-cell current-clamp recordings from a local astrocyte. This approach takes advantage of the fact that the low input resistance of electrically passive astroglia allows extracellular currents to pass through the astrocytic membrane with relatively little attenuation. Once the slice preparation is ready, it takes ∼30 min to several hours to implement this protocol, depending on the experimental design, which is similar to other patch-clamp techniques. The technique presented here can be used to directly access the intracellular medium of individual astrocytes while examining synapses functioning in their immediate proximity.

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Figure 1: Whole-cell recordings using an astrocyte patch pipette are sensitive to extracellular field potentials.
Figure 2: The amplitude of field responses recorded through an astrocytic whole-cell pipette (a-fEPSPs) faithfully represents the initial slope of fEPSPs recorded with a standard extracellular electrode.
Figure 3: The electrophysiological makeup of a-fEPSPs makes them suitable for representing extracellular fEPSPs.
Figure 4: The a-fEPSP amplitude measured using an astrocyte pipette robustly represents extracellular fEPSPs during long-term potentiation.
Figure 5: Testing the suitability of other a-fEPSP components to represent the classical fEPSP initial slope measure.
Figure 6: Individual astrocytes influence LTP induction mainly at nearby synapses: an experiment enabled by the recording of local synaptic activity through an astrocytic patch pipette.

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Acknowledgements

This work was supported by the Human Frontier Science Programme, the Wellcome Trust (D.A.R.), the Medical Research Council (D.A.R.), a UCL Excellence Fellowship (C.H.) and the NRW-Rückkehrerprogramm (C.H.).

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Authors and Affiliations

  1. UCL Institute of Neurology, University College London, London, UK

    Christian Henneberger & Dmitri A Rusakov

  2. Institute of Cellular Neurosciences, University of Bonn Medical School, Bonn, Germany

    Christian Henneberger

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  1. Christian Henneberger
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  2. Dmitri A Rusakov
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C.H. carried out experimental studies; D.A.R. and C.H. designed the study, analyzed the data and wrote the paper.

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Correspondence to Christian Henneberger or Dmitri A Rusakov.

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Henneberger, C., Rusakov, D. Monitoring local synaptic activity with astrocytic patch pipettes. Nat Protoc 7, 2171–2179 (2012). https://doi.org/10.1038/nprot.2012.140

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