Abstract

The zebrafish is an established model to study the development and function of visual neuronal circuits in vivo, largely due to their optical accessibility at embryonic and larval stages. In the past decade multiple experimental paradigms have been developed to study visually-driven behaviours, particularly those regulated by the optic tectum, the main visual centre in lower vertebrates. With few exceptions these techniques are limited to young larvae (7–9 days post-fertilisation, dpf). However, many forms of visually-driven behaviour, such as shoaling, emerge at later developmental stages. Consequently, there is a need for an experimental paradigm to image the visual system in zebrafish larvae beyond 9 dpf. Here, we show that using NBT:GCaMP3 line allows for imaging neuronal activity in the optic tectum in late stage larvae until at least 21 dpf. Utilising this line, we have characterised the receptive field properties of tectal neurons of the 2–3 weeks old fish in the cell bodies and the neuropil. The NBT:GCaMP3 line provides a complementary approach and additional opportunities to study neuronal activity in late stage zebrafish larvae.

Highlights

  • The zebrafish has become an important model organism to study the development and organisation of neuronal circuits involved in various forms of behaviour

  • The commonly used huc:GCaMP3 reporter line decreases in brightness after 7–9 dpf [12]

  • GCaMP3 [27] under the control of the Xenopus neuronal beta-tubulin (NBT) promoter (Materials and Methods, [26]). This promoter has been sub-cloned from Xenopus laevis, where it normally drives the

Read more

Summary

Introduction

The zebrafish has become an important model organism to study the development and organisation of neuronal circuits involved in various forms of behaviour. Many experimental paradigms combining behavioural assays with imaging of neuronal activity have been developed and applied to study the tuning properties of visual neurons in the retina, optic tectum, pretectum, and habenula [1,2,3,4,5,6,7]. A disadvantage of the commonly used pan-neuronal huc promoter is that the expression of neuronal activity reporters (e.g., GCaMPs) decreases after 6–7 dpf [12]. To overcome this issue, one can either use GCaMP3-H2B fusion to target the calcium reporter expression to the nucleus [12]

Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call