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Expression of Channelrhodopsin-2 Using in Suspension Electroporation for Studying the Monosynaptic Transmission in Neuronal Culture

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Abstract

Understanding the mechanism of synaptic transmission and its plasticity is one of the central goals of modern neuroscience. Neuronal culture is a very convenient model for studying mechanisms of synaptic transmission. However, investigation of two or more synaptically connected neurons in culture by conventional methods is a difficult task. In this study, we describe new protocol for studying the synaptic transmission between cultured neurons using in suspension electroporation for the expression of channelrhodopsin-2 (ChR2) which was applied to only certain subpopulation of cultured cells, leaving the majority of neurons completely untreated. We show that this technique allows reliable long-lasting (hours) recording of monosynaptic excitatory postsynaptic potentials (EPSPs) in cultured hippocampal neurons using a repeated light stimulation of neighboring ChR2-expressing neurons.

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Acknowledgments

This research was funded by the Russian Science Foundation grant 14-25-00072 (neuronal cell culture and development of the electroporation protocol) and by RFBR grant no. 15-04-06286 (whole cell recording with optical stimulation).

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Authors and Affiliations

  1. Institute of Higher Nervous Activity and Neurophysiology of RAS, 5A Butlerova St., Moscow, 117485, Russia

    Natalia Bal, Aleksey Malyshev, Ivan Smirnov & Pavel Balaban

Authors
  1. Natalia Bal
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  2. Aleksey Malyshev
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  3. Ivan Smirnov
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  4. Pavel Balaban
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Corresponding author

Correspondence to Natalia Bal.

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All experimental procedures were conducted in accordance with the European Communities Council Directive of 24 November 1986 (86/609/ EEC) on the protection of animals used for scientific purposes. The study protocol was approved by the Ethics Committee of the Institute of Higher Nervous Activity and Neurophysiology of RAS.

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Bal, N., Malyshev, A., Smirnov, I. et al. Expression of Channelrhodopsin-2 Using in Suspension Electroporation for Studying the Monosynaptic Transmission in Neuronal Culture. BioNanoSci. 6, 329–331 (2016). https://doi.org/10.1007/s12668-016-0228-7

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  • DOI: https://doi.org/10.1007/s12668-016-0228-7

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