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Sci Rep. 2018 Apr 16;8(1):6048. doi: 10.1038/s41598-018-23906-1.

Optical interrogation of neuronal circuitry in zebrafish using genetically encoded voltage indicators.

Author information

1
Division of Life Science, Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan.
2
Riken Brain Science Institute, Hirosawa, Wako City, Saitama, 351-0198, Japan.
3
Department of Molecular and Cellular Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA, 02138, USA.
4
Saitama University Brain Science Institute, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan.
5
Division of Life Science, Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan. stsuda@mail.saitama-u.ac.jp.
6
Saitama University Brain Science Institute, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan. stsuda@mail.saitama-u.ac.jp.
7
Research and Development Bureau, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama City, Saitama, 338-8570, Japan. stsuda@mail.saitama-u.ac.jp.

Abstract

Optical measurement of membrane potentials enables fast, direct and simultaneous detection of membrane potentials from a population of neurons, providing a desirable approach for functional analysis of neuronal circuits. Here, we applied recently developed genetically encoded voltage indicators, ASAP1 (Accelerated Sensor of Action Potentials 1) and QuasAr2 (Quality superior to Arch 2), to zebrafish, an ideal model system for studying neurogenesis. To achieve this, we established transgenic lines which express the voltage sensors, and showed that ASAP1 is expressed in zebrafish neurons. To examine whether neuronal activity could be detected by ASAP1, we performed whole-cerebellum imaging, showing that depolarization was detected widely in the cerebellum and optic tectum upon electrical stimulation. Spontaneous activity in the spinal cord was also detected by ASAP1 imaging at single-cell resolution as well as at the neuronal population level. These responses mostly disappeared following treatment with tetrodotoxin, indicating that ASAP1 enabled optical measurement of neuronal activity in the zebrafish brain. Combining this method with other approaches, such as optogenetics and behavioural analysis may facilitate a deeper understanding of the functional organization of brain circuitry and its development.

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