Abstract
Visualization of neuronal activity during animal behavior is a critical step in understanding how the brain generates behavior. In the model vertebrate zebrafish, imaging of the brain has been done mostly by using immobilized fish. Here, we describe a novel method to image neuronal activity of the larval zebrafish brain during prey capture behavior. We expressed a genetically encoded fluorescent calcium indicator, GCaMP, in the optic tectum of the midbrain using the Gal4-UAS system. Tectal activity was then imaged in unrestrained larvae during prey perception. Since larval zebrafish swim only intermittently, detection of the neuronal activity is possible between swimming bouts. Our method makes functional brain imaging under natural behavioral conditions feasible and will greatly benefit the study of neuronal activities that evoke animal behaviors.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Kettunen P (2012) Calcium imaging in the zebrafish. Adv Exp Med Biol 740:1039–1071
Semmelhack JL et al (2015) A dedicated visual pathway for prey detection in larval zebrafish. eLife 2014;3:e04878
Ritter DA, Bhatt DH, Fetcho JR (2001) In vivo imaging of zebrafish reveals differences in the spinal networks for escape and swimming movements. J Neurosci 21(22):8956–8965
Bianco IH, Engert F (2015) Visuomotor transformations underlying hunting behavior in zebrafish. Curr Biol 25(7):831–846
Nakai J, Ohkura M, Imoto K (2001) A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein. Nat Biotechnol 19(2):137–141
Muto A et al (2013) Real-time visualization of neuronal activity during perception. Curr Biol 23(4):307–311
Muto A et al (2011) Genetic visualization with an improved GCaMP calcium indicator reveals spatiotemporal activation of the spinal motor neurons in zebrafish. Proc Natl Acad Sci U S A 108(13):5425–5430
Asakawa K, Kawakami K (2008) Targeted gene expression by the Gal4-UAS system in zebrafish. Dev Growth Differ 50(6):391–399
Asakawa K et al (2008) Genetic dissection of neural circuits by Tol2 transposon-mediated Gal4 gene and enhancer trapping in zebrafish. Proc Natl Acad Sci U S A 105(4):1255–1260
Kawakami K et al (2010) zTrap: zebrafish gene trap and enhancer trap database. BMC Dev Biol 10:105
Lister JA et al (1999) Nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate. Development 126(17):3757–3767
Acknowledgments
This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) in Japan: KAKENHI Grant Number 24120521, 25290009, and 25650120.
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
Muto, A., Kawakami, K. (2016). Calcium Imaging of Neuronal Activity in Free-Swimming 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_23
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3771-4_23
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