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Real Time and Repeated Measurement of Skeletal Muscle Growth in Individual Live Zebrafish Subjected to Altered Electrical Activity

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Original languageEnglish
Article numbere64063
Pages (from-to)1-23
Number of pages23
JournalJournal of visualized experiments : JoVE
Issue number184
Accepted/In press26 May 2022
Published16 Jun 2022

Bibliographical note

Funding Information: The authors are deeply indebted to the efforts of Hughes lab members Drs Seetharamaiah Attili, Jana Koth, Fernanda Bajanca, Victoria C. Williams, Yaniv Hinits, Giorgia Bergamin, and Vladimir Snetkov for development of the described protocols, and to Henry Roehl, Christina Hammond, David Langenau and Peter Currie for sharing plasmids or zebrafish lines. SMH is a Medical Research Council (MRC) Scientist with Programme Grant G1001029, MR/N021231/1, and MR/W001381/1support. MA held a MRC Doctoral Training Programme PhD Studentship from King's College London. This work benefited from the trigonometrical input of David M. Robinson, scholar, mentor, and friend. Publisher Copyright: © 2022 JoVE.


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King's Authors


A number of methods can be used to visualize individual cells throughout the body of
live embryonic, larval or juvenile zebrafish. We show that live fish with fluorescentlymarked
plasma membranes can be scanned in a confocal laser scanning microscope
in order to determine the volume of muscle tissue and the number of muscle fibers
present. Efficient approaches for the measurement of cell number and size in live
animals over time are described and validated against more arduous segmentation
methods. Methods are described that permit the control of muscle electrical, and
thus contractile, activity. Loss of skeletal muscle contractile activity greatly reduced
muscle growth. In larvae, a protocol is described that allows reintroduction of
patterned electrical-evoked contractile activity. The described methods minimize the
effect of inter-individual variability and will permit analysis of the effect of electrical,
genetic, drug, or environmental stimuli on a variety of cellular and physiological
growth parameters in the context of the living organism. Long-term follow-up of the
measured effects of a defined early-life intervention on individuals can subsequently
be performed.

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