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Neuroscience
成年斑马鱼前脑活动的双光子钙成像
成年斑马鱼前脑活动的双光子钙成像
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Two-Photon Calcium Imaging of Forebrain Activity in Behaving Adult Zebrafish

成年斑马鱼前脑活动的双光子钙成像

Full Text
3,750 Views
07:21 min
July 28, 2023

DOI: 10.3791/65526-v

Jose Sandino Bandonil*1,2, Yi-Hsuan Liao*1, Amir Fathi1, Kuo-Hua Huang1,2

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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Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

在这里,我们提出了一种在成年斑马鱼的背前脑中进行双光子钙成像的方案。

鱼类前脑所在的区域被怀疑与参与复杂行为的哺乳动物大脑区域同源。这些包括海马体、皮层和杏仁核。我们目前正在试图了解感觉信息对这些大脑区域活动的影响。

目前斑马鱼神经科学的研究使用固定幼虫的双光子钙成像。结合基于虚拟现实的行为论文,研究大脑中的感觉运动处理。大多数关于斑马鱼大脑的研究都是在幼虫上进行的,但是鱼类的一些复杂行为直到成年才完全发育。

我们的协议允许研究成人在行为过程中的大脑活动。与以前的方法相比,该方案将手术时间缩短了 50%。这是一种更有效的方法,可以限制成年斑马鱼的头部以获得更好的图像。

这种头部约束方案为在成年斑马鱼大脑上进行体内实验提供了机会。除了双光子钙图像外,头部约束还可能允许对活鱼进行光遗传学操作、电生理学甚至超声成像。

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