Abstract
We present a protocol to combine behavioral recording and imaging using 2-photon laser-scanning microscopy in head-fixed larval zebrafish that express a genetically encoded calcium indicator. The steps involve restraining the larva in agarose, setting up optics that allow projection of a visual stimulus and infrared illumination to monitor behavior, and analysis of the neuronal and behavioral data.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fetcho JR, O’Malley DM (1997) Imaging neuronal networks in behaving animals. Curr Opin Neurobiol 7:832–838
Kettunen P (2012) Calcium imaging in the zebrafish. Adv Exp Med Biol 740:1039–1071
Ahrens MB, Li JM, Orger MB et al (2012) Brain-wide neuronal dynamics during motor adaptation in zebrafish. Nature 485:471–477
Ahrens MB, Orger MB, Robson DN et al (2013) Whole-brain functional imaging at cellular resolution using light-sheet microscopy. Nat Methods 10:413–420
Wolf S, Supatto W, Debrégeas G et al (2015) Whole-brain functional imaging with two-photon light-sheet microscopy. Nat Methods 12:379–380
Portugues R, Feierstein CE, Engert F et al (2014) Whole-brain activity maps reveal stereotyped, distributed networks for visuomotor behavior. Neuron 81:1328–1343
Renninger SL, Orger MB (2013) Two-photon imaging of neural population activity in zebrafish. Methods 62:255–267
Niell CM, Smith SJ (2005) Functional imaging reveals rapid development of visual response properties in the zebrafish tectum. Neuron 45:941–951
O'Malley DM, Sankrithi NS, Borla MA et al (2004) Optical physiology and locomotor behaviors of wild-type and nacre zebrafish. Methods Cell Biol 76:261–284
Portugues R, Engert F (2011) Adaptive locomotor behavior in larval zebrafish. Front Syst Neurosci 5:72
Lister JA, Robertson CP, Lepage T et al (1999) Nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate. Development 126:3757–3767
Severi K, Portugues R, Marques J et al (2014) Neural control and modulation of swimming speed in the larval zebrafish. Neuron 83(3):692–707
Orger MB, Kampff AR, Severi KE et al (2008) Control of visually guided behavior by distinct populations of spinal projection neurons. Nat Neurosci 11:327–333
Miri A, Daie K, Burdine RD et al (2011) Regression-based identification of behavior-encoding neurons during large-scale optical imaging of neural activity at cellular resolution. J Neurophysiol 105:964–980
Nestares O, Heeger DJ (2000) Robust multiresolution alignment of MRI brain volumes. Magn Reson Med 43:705–715
Mukamel EA, Nimmerjahn A, Schnitzer MJ (2009) Automated analysis of cellular signals from large-scale calcium imaging data. Neuron 63:747–760
Romano SA, Pietri T, Pérez-Schuster V et al (2015) Spontaneous neuronal network dynamics reveal circuit’s functional adaptations for behavior. Neuron 85:1070–1085
Kim CK, Miri A, Leung LC et al (2014) Prolonged, brain-wide expression of nuclear-localized GCaMP3 for functional circuit mapping. Front Neural Circ 8:138
Akerboom J, Chen TW, Wardill TJ et al (2012) Optimization of a GCaMP calcium indicator for neural activity imaging. J Neurosci 32:13819–13840
Straw AD (2008) Vision egg: an open-source library for realtime visual stimulus generation. Front Neuroinformatics 2
Brainard DH (1997) The psychophysics toolbox. Spat Vis 10:433–436
Page-Mccaw PS, Chung SC, Muto A et al (2004) Retinal network adaptation to bright light requires tyrosinase. Nat Neurosci 7:1329–1336
Kubo F, Hablitzel B, Maschio MD et al (2014) Functional architecture of an optic flow-responsive area that drives horizontal eye movements in zebrafish. Neuron 81:1344–1359
Bianco IH, Engert F (2015) Visuomotor transformations underlying hunting behavior in zebrafish. Curr Biol 25:831–846
Huber D, Gutnisky DA, Peron S et al (2012) Multiple dynamic representations in the motor cortex during sensorimotor learning. Nature 484:473–478
Acknowledgments
The authors would like to acknowledge the contribution of colleagues Claudia Feierstein and Florian Engert to the development of the methods described in this protocol. MBO was supported by a Marie Curie Career Integration Grant, PCIG09-GA-2011-294049. RP was supported by the Max Planck Society.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Orger, M.B., Portugues, R. (2016). Correlating Whole Brain Neural Activity with Behavior in Head-Fixed Larval Zebrafish. In: Kawakami, K., Patton, E., Orger, M. (eds) Zebrafish. Methods in Molecular Biology, vol 1451. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3771-4_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3771-4_21
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3769-1
Online ISBN: 978-1-4939-3771-4
eBook Packages: Springer Protocols