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Review
. 2023 Aug;18(8):2415-2440.
doi: 10.1038/s41596-023-00842-7. Epub 2023 Jul 7.

Opto-juxtacellular interrogation of neural circuits in freely moving mice

Lingjun Ding #  1   2   3 , Giuseppe Balsamo #  1   2   3 , Maria Diamantaki  1   2   3   4 , Patricia Preston-Ferrer  5   6 , Andrea Burgalossi  7   8
Affiliations
Review

Opto-juxtacellular interrogation of neural circuits in freely moving mice

Lingjun Ding et al. Nat Protoc. 2023 Aug.

Abstract

Neural circuits are assembled from an enormous variety of neuronal cell types. Although significant advances have been made in classifying neurons on the basis of morphological, molecular and electrophysiological properties, understanding how this diversity contributes to brain function during behavior has remained a major experimental challenge. Here, we present an extension to our previous protocol, in which we describe the technical procedures for performing juxtacellular opto-tagging of single neurons in freely moving mice by using Channelrhodopsin-2-expressing viral vectors. This method allows one to selectively target molecularly defined cell classes for in vivo single-cell recordings. The targeted cells can be labeled via juxtacellular procedures and further characterized via post-hoc morphological and molecular analysis. In its current form, the protocol allows multiple recording and labeling attempts to be performed within individual animals, by means of a mechanical pipette micropositioning system. We provide proof-of-principle validation of this technique by recording from Calbindin-positive pyramidal neurons in the mouse hippocampus during spatial exploration; however, this approach can easily be extended to other behaviors and cortical or subcortical areas. The procedures described here, from the viral injection to the histological processing of brain sections, can be completed in ~4-5 weeks.This protocol is an extension to: Nat. Protoc. 9, 2369-2381 (2014): https://doi.org/10.1038/nprot.2014.161.

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