Two-Photon Calcium Imaging of Forebrain Activity in Behaving Adult Zebrafish

Jose Sandino Bandonil; Yi-Hsuan Liao; Amir Fathi; Kuo-Hua Huang

doi:10.3791/65526

Imagerie calcique à deux photons de l’activité du cerveau antérieur chez le poisson-zèbre adulte

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Neuroscience

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JoVE Journal Neuroscience

Two-Photon Calcium Imaging of Forebrain Activity in Behaving Adult Zebrafish

Imagerie calcique à deux photons de l’activité du cerveau antérieur chez le poisson-zèbre adulte

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07:21 min

July 28, 2023

DOI: 10.3791/65526-v

Jose Sandino Bandonil*^1,2, Yi-Hsuan Liao*¹, Amir Fathi¹, Kuo-Hua Huang^1,2

¹Institute of Molecular Biology,Academia Sinica, ²Molecular and Cell Biology, Taiwan International Graduate Program, Academia Sinica and Graduate Institute of Life Sciences,National Defense Medical Center

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Overview

This study presents a protocol for two-photon calcium imaging to investigate brain activity in the dorsal forebrain of adult zebrafish. The approach allows researchers to examine the effects of sensory information on neuronal activity associated with complex behaviors, overcoming limitations faced in larval studies.

Key Study Components

Area of Science

Neuroscience
Imaging Techniques
Behavioral Neuroscience

Background

The zebrafish forebrain has regions homologous to mammalian areas involved in behavior.
Previous research primarily focused on larval stages, leaving adult complexities underexplored.
This protocol aims to fill knowledge gaps regarding adult zebrafish brain activity.

Purpose of Study

To develop an efficient method to study brain activity in adult zebrafish.
To facilitate the simultaneous examination of sensory information impact on behavior.
To reduce surgery time and enhance imaging capabilities.

Methods Used

Two-photon calcium imaging using a head-restrained adult zebrafish model.
Head restraint enabled imaging of up to 200 microns deep in the dorsal forebrain.
Steps included anesthesia, head bar attachment, and surgical preparation for imaging.
Behavioral recording was integrated with calcium imaging during experiments.

Main Results

The method allows visualization of individual neuronal activity in complex behavioral contexts.
Key findings include enhanced access to pallial areas of the zebrafish brain.
Simultaneous imaging and recording reveal neuronal correlates of motor outputs.

Conclusions

This study enables more in-depth investigation of adult zebrafish brain activity during behavior.
The protocol contributes to understanding sensory-motor processing and neuronal mechanisms.

Frequently Asked Questions

What is the advantage of using adult zebrafish over larvae?

Adult zebrafish exhibit more complex behaviors that are essential for understanding certain neurobiological processes, which are not fully developed in larvae.

How is the head restraint method implemented?

The method involves surgical attachment of a head bar to immobilize the fish, which allows for precise imaging techniques while minimizing surgery time.

What type of data is obtained from this imaging technique?

The two-photon calcium imaging provides real-time recordings of neuronal activity in response to sensory stimuli and during behavioral assays.

How can this protocol be adapted for other studies?

This imaging technique can be integrated with optogenetic manipulation or other imaging methods to study different aspects of brain function and behavior.

What are the critical steps of the surgical procedure?

Critical steps include anesthesia, head bar attachment, and preparation of the skull for optimal imaging access.

Are there any limitations to this method?

The protocol may require significant technical skill and careful handling of the fish to avoid distress during surgery.

Nous présentons ici un protocole permettant d’effectuer une imagerie calcique à deux photons dans le cerveau antérieur dorsal du poisson-zèbre adulte.

Le cerveau antérieur du poisson héberge des zones soupçonnées d’être homologues aux régions cérébrales des mammifères impliquées dans un comportement complexe. Il s’agit notamment de l’hippocampe, du cortex et de l’amygdale. Nous essayons actuellement de comprendre l’effet de l’information sensorielle sur l’activité de ces régions du cerveau.

Les recherches actuelles en neurosciences du poisson zèbre utilisent l’imagerie calcique à deux photons de larves immobilisées. Combiné avec des essais de comportement basés sur la réalité virtuelle, pour étudier le traitement sensori-moteur dans le cerveau. La plupart des recherches sur le cerveau du poisson zèbre sont effectuées sur des larves, mais certains comportements complexes chez les poissons ne se développent pas complètement avant l’âge adulte.

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Neurosciences Numéro 197 Poisson-zèbre adulte Comportement Fonctions cognitives Méthodes d’imagerie optique Enregistrement de l’activité cérébrale Cerveau antérieur dorsal Chirurgie de l’appuie-tête Enregistrement des neurones Environnement visuel immersif Comportements déclenchés visuellement