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J Neurosci Methods. 2014 Sep 30;235:83-91. doi: 10.1016/j.jneumeth.2014.06.031. Epub 2014 Jul 6.

Investigating local and long-range neuronal network dynamics by simultaneous optogenetics, reverse microdialysis and silicon probe recordings in vivo.

Author information

1
Neuroscience Division, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK.
2
Neuroscience Division, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK; Department of Pharmacology, Faculty of Pharmacy, Instanbul University, Istanbul, Turkey.
3
Department of Pharmacology and Clinical Pharmacology, Faculty of Medicine, Marmara University, Istanbul, Turkey.
4
Neuroscience Division, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK; Department of Biochemistry and Physiology, University of Malta, Malta.
5
Sorbonne Universités, UPMC Univ Paris 06, UM CR18, Neuroscience Paris Seine (NPS), Paris F-75005, France; CNRS, UMR 8246, NPS, Paris F-75005, France.
6
Neuroscience Division, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK. Electronic address: crunelli@cardiff.ac.uk.
7
Neuroscience Division, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, UK. Electronic address: davidfo@cardiff.ac.uk.

Abstract

BACKGROUND:

The advent of optogenetics has given neuroscientists the opportunity to excite or inhibit neuronal population activity with high temporal resolution and cellular selectivity. Thus, when combined with recordings of neuronal ensemble activity in freely moving animals optogenetics can provide an unprecedented snapshot of the contribution of neuronal assemblies to (patho)physiological conditions in vivo. Still, the combination of optogenetic and silicone probe (or tetrode) recordings does not allow investigation of the role played by voltage- and transmitter-gated channels of the opsin-transfected neurons and/or other adjacent neurons in controlling neuronal activity.

NEW METHOD AND RESULTS:

We demonstrate that optogenetics and silicone probe recordings can be combined with intracerebral reverse microdialysis for the long-term delivery of neuroactive drugs around the optic fiber and silicone probe. In particular, we show the effect of antagonists of T-type Ca(2+) channels, hyperpolarization-activated cyclic nucleotide-gated channels and metabotropic glutamate receptors on silicone probe-recorded activity of the local opsin-transfected neurons in the ventrobasal thalamus, and demonstrate the changes that the block of these thalamic channels/receptors brings about in the network dynamics of distant somatotopic cortical neuronal ensembles.

COMPARISON WITH EXISTING METHODS:

This is the first demonstration of successfully combining optogenetics and neuronal ensemble recordings with reverse microdialysis. This combination of techniques overcomes some of the disadvantages that are associated with the use of intracerebral injection of a drug-containing solution at the site of laser activation.

CONCLUSIONS:

The combination of reverse microdialysis, silicone probe recordings and optogenetics can unravel the short and long-term effects of specific transmitter- and voltage-gated channels on laser-modulated firing at the site of optogenetic stimulation and the actions that these manipulations exert on distant neuronal populations.

KEYWORDS:

Cortex; Delta waves; EEG; HCN channels; Metabotropic glutamate receptors; Sleep spindles; Slow waves; T-type Ca(2+) channels; Thalamus

PMID:
25004203
PMCID:
PMC4164909
DOI:
10.1016/j.jneumeth.2014.06.031
[Indexed for MEDLINE]
Free PMC Article

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