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Front. Neural Circuits | doi: 10.3389/fncir.2018.00033

Two-photon functional imaging of the auditory cortex in behaving mice: From neural networks to single spines

  • 1Army Medical University, China
  • 2Institute of Urinary Surgery, Southwest Hospital, Third Military Medical University, China
  • 3Brain Research Instrument Innovation Center (BRIIC), Suzhou Institute of Biomedical Engineering and Technology (CAS), China
  • 4CAS Center for Excellence in Brain Science and Intelligence Technology (CEBSIT), Chinese Academy of Sciences, China

In vivo two-photon Ca2+ imaging is a powerful tool for recording neuronal activities during perceptual tasks and has been increasingly applied to behaving animals for acute or chronic experiments. However, the auditory cortex is not easily accessible to imaging because of the abundant temporal muscles, arteries around the ears and their lateral locations. Here, we report a protocol for two-photon Ca2+ imaging in the auditory cortex of head-fixed behaving mice. By using a custom-made head fixation apparatus and a head-rotated fixation procedure, we achieved two-photon imaging and in combination with targeted cell-attached recordings of auditory cortical neurons in behaving mice. Using synthetic Ca2+ indicators, we recorded the Ca2+ transients at multiple scales, including neuronal populations, single neurons, dendrites and single spines, in auditory cortex during behavior. Furthermore, using genetically encoded Ca2+ indicators (GECIs), we monitored the neuronal dynamics over days throughout the process of associative learning. Therefore, we achieved two-photon functional imaging at multiple scales in auditory cortex of behaving mice, which extends the tool box for investigating the neural basis of audition-related behaviors.

Keywords: two-photon Ca2+ imaging, Auditory Cortex, behaving mouse, Dendritic Spines, cell-attached recordings

Received: 23 Dec 2017; Accepted: 10 Apr 2018.

Edited by:

Daniel B. Polley, Massachusetts Eye & Ear Infirmary, Harvard Medical School, United States

Reviewed by:

Gunsoo Kim, CNIR-IBS
John Issa, Northwestern University, United States  

Copyright: © 2018 Li, Wang, Yao, Liang, Liao, Yang, Zhang, Yan, Jia, Chen and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Prof. Xiaowei Chen, Army Medical University, Chongqing, China,
PhD. Xingyi Li, Army Medical University, Chongqing, China,