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. 2023 Dec 15;4(4):102517.
doi: 10.1016/j.xpro.2023.102517. Epub 2023 Sep 23.

A protocol for whole-brain Ca2+ imaging in Astyanax mexicanus, a model of comparative evolution

Evan Lloyd  1 , Martin Privat  2 , German Sumbre  2 , Erik R Duboué  3 , Alex C Keene  4
Affiliations

A protocol for whole-brain Ca2+ imaging in Astyanax mexicanus, a model of comparative evolution

Evan Lloyd et al. STAR Protoc. .

Abstract

In this protocol, we describe a comparative approach to study the evolution of brain function in the Mexican tetra, Astyanax mexicanus. We developed surface fish and two independent populations of cavefish with pan-neuronal expression of the Ca2+ sensor GCaMP6s. We describe a methodology to prepare samples and image activity across the optic tectum and olfactory bulb.

Keywords: Evolutionary Biology; Neuroscience.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Husbandry and genetics (A) Timeline of breeding and screening for transgenic animals. (B) Examples of healthy live embryos (left), and unviable embryos (right).
Figure 2
Figure 2
Functional imaging of light response in the tectum (A) The visual imaging system. Inset: a larva mounted in agarose, with white light LED system arranged in the field-of-view of the larva. (B) Results of post-processing of visual imaging; a surface fish optic tectum, with individual neurons programmatically segmented (left); representative fluorescence traces of neurons during visual stimulation, with significant Ca2+ transients highlighted in red (right).
Figure 3
Figure 3
Functional imaging of olfactory response in the olfactory bulb (A) The olfactory imaging system. Inset: a close-up view of the larvae, mounted in agarose, with odor inflow on the left, and outflow chamber on the right. (B) Results of post-processing of olfactory imaging; a cavefish olfactory bulb with individual neurons programmatically segmented (left); representative fluorescence traces of neurons during olfactory stimulation, with significant Ca2+ transients highlighted in red (right).
See this image and copyright information in PMC

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