A Versatile Method for Viral Transfection of Calcium Indicators in the Neonatal Mouse Brain
Abstract
The first three postnatal weeks in rodents are a time when sensory experience drives the maturation of brain circuits, an important process that is not yet well understood. Alterations in this critical period of experience-dependent circuit assembly and plasticity contribute to several neurodevelopmental disorders, such as autism, epilepsy, and schizophrenia. Therefore, techniques for recording network activity and tracing neuronal connectivity over this time period are necessary for delineating circuit refinement in typical development and how it deviates in disease. Calcium imaging with GCaMP6 and other genetically encoded indicators is rapidly becoming the preferred method for recording network activity at the single-synapse and single-cell level in vivo, especially in genetically identified neuronal populations. We describe a protocol for intracortical injection of recombinant adeno-associated viruses in P1 neonatal mice and demonstrate its use for longitudinal imaging of GCaMP6s in the same neurons over several weeks to characterize the developmental desynchronization of cortical network activity. Our approach is ideally suited for chronic in vivo two-photon calcium imaging of neuronal activity from synapses to entire networks during the early postnatal period.
Additional Information
© 2018 He, Arroyo, Cantu, Goel and Portera-Cailliau. 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(s) 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. Received: 04 April 2018; Accepted: 29 June 2018; Published: 23 July 2018. This research was supported by a Paul & Daisy Soros Fellowship for New Americans and NIH NINDS F30 Fellowship NS093719 to CH, and the UCLA Medical Scientist Training Program (NIH NIGMS training grant T32 GM008042); a UCLA dissertation year fellowship and the UCLA Neurobehavioral Genetics Training Grant (T32 5T32MH073526-08) to EA; Eugene V. Cota-Robles Fellowships to EA and DC; the UCLA Neural Microcircuits Training Grant (T32-NS058280) to DC; W81XWH-14-1-0433 (USAMRMC, DOD), a Developmental Disabilities Translational Research Program grant #20160969 (The John Merck Fund), SFARI grant 295438 (Simons Foundation) and NIH NICHD grant R01 HD054453 to CP-C. We thank the Janelia GENIE project for the AAV-Syn-GCaMP6s, D. Buonomano for his assistance in our electrophysiological experiments, and P. Mineault, D. Ringach, D. Dombeck, T.-W. Chen, and K. Svoboda for MATLAB code used for imaging analysis. We also thank K. Battista for the P1 injection illustration (Figure 1) and N. Kourdougli for assistance with Figure 3. Data Availability Statement: All datasets generated and analyzed for this study are available upon request to the corresponding author. Author Contributions: CH and CP-C conceived the project and designed the experiments. CH and DC developed the P1 injection and imaging protocols and wrote the MATLAB code. CH, EA, and AG conducted the experiments and analyzed the data. CH, EA, and CP-C wrote the paper. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.Attached Files
Published - fncir-12-00056.pdf
Supplemental Material - 4174163.zip
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Additional details
- PMCID
- PMC6064716
- Eprint ID
- 88471
- Resolver ID
- CaltechAUTHORS:20180801-141010696
- Paul and Daisy Soros Fellowship
- F30 NS093719
- NIH Postdoctoral Fellowship
- T32 GM008042
- NIH Predoctoral Fellowship
- W81XWH-14-1-0433
- Army Medical Research and Materiel Command
- 20160969
- John Merck Fund
- 295438
- Simons Foundation
- UCLA
- 5T32MH073526-08
- NIH Predoctoral Fellowship
- T32-NS058280
- NIH Predoctoral Fellowship
- R01 HD054453
- NIH
- Created
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2018-08-01Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field