Monitoring neural activity with bioluminescence during natural behavior
Abstract
Existing techniques for monitoring neural activity in awake, freely behaving vertebrates are invasive and difficult to target to genetically identified neurons. We used bioluminescence to non-invasively monitor the activity of genetically specified neurons in freely behaving zebrafish. Transgenic fish with the Ca^(2+)-sensitive photoprotein green fluorescent protein (GFP)-Aequorin in most neurons generated large and fast bioluminescent signals that were related to neural activity, neuroluminescence, which could be recorded continuously for many days. To test the limits of this technique, we specifically targeted GFP-Aequorin to the hypocretin-positive neurons of the hypothalamus. We found that neuroluminescence generated by this group of ~20 neurons was associated with periods of increased locomotor activity and identified two classes of neural activity corresponding to distinct swim latencies. Our neuroluminescence assay can report, with high temporal resolution and sensitivity, the activity of small subsets of neurons during unrestrained behavior.
Additional Information
© 2010 Nature Publishing Group. Received 9 November 2009; accepted 22 February 2010; published online 21 March 2010. We thank W. Hastings and T. Wilson for bountiful advice and discussion and generously providing an intensified CCD camera. We also thank L. Tricoire for the kind gift of the GFP-apoAequorin construct, M. Orger, A. Douglass, P. Ramdya, and members of the Engert and Schier laboratories for comments and advice, A. Douglass for Nβt-gal4 vectors, P. Ramdya for providing the nacre strains and B. Obama for his stimulation package. We thank S. Zimmerman, K. Hurley, and J. Miller for excellent zebrafish care. This work was funded by the McKnight Foundation (F.E.), the Harvard Mind, Brain and Behavior post-doctoral fellows program (A.R.K.), and the US National Institutes of Health (A.F.S. and D.A.P.). Author Contributions: E.A.N. and A.R.K. designed the assay and performed the experiments. E.A.N., A.R.K. and F.E. analyzed the data. D.A.P. and A.F.S. generated the HCRT–GFP-apoAequorin transgenic line and assisted with behavioral analysis. E.A.N., A.R.K., D.A.P., A.F.S. and F.E. prepared the manuscript. E.A.N. suffered the most.Attached Files
Published - NatNeuro.pdf
Accepted Version - nihms181675.pdf
Supplemental Material - nn.2518-S1.pdf
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Additional details
- PMCID
- PMC2846983
- Eprint ID
- 17944
- Resolver ID
- CaltechAUTHORS:20100412-134749684
- McKnight Foundation
- Harvard Mind, Brain and Behavior post-doctoral fellows
- NIH
- Created
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2010-04-30Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field