HuC-eGFP mosaic labelling of neurons in zebrafish enables in vivo live cell imaging of growth cones

St John, James A. and Key, Brian (2012) HuC-eGFP mosaic labelling of neurons in zebrafish enables in vivo live cell imaging of growth cones. Journal of Molecular Histology, 43 6: 615-623. doi:10.1007/s10735-012-9462-7

Author St John, James A.
Key, Brian
Title HuC-eGFP mosaic labelling of neurons in zebrafish enables in vivo live cell imaging of growth cones
Journal name Journal of Molecular Histology   Check publisher's open access policy
ISSN 1567-2379
Publication date 2012-12
Sub-type Article (original research)
DOI 10.1007/s10735-012-9462-7
Volume 43
Issue 6
Start page 615
End page 623
Total pages 9
Place of publication Dordrecht, The Netherlands
Publisher Springer Netherlands
Collection year 2013
Language eng
Formatted abstract
The field of axon guidance is taking advantage of the powerful genetic and imaging tools that are now available to visualise growth behaviour in living cells, both in vivo and in real time. We have developed a method to visualise individual neurons within the living zebrafish embryo which provides exceptional cellular resolution of growth cones and their filopodia. We generated a DNA construct in which the HuC promoter drives expression of eGFP. Injection of the plasmid into single cell fertilised zebrafish egg resulted in mosaic expression of eGFP in neurons throughout the developing embryo. By manipulating the concentration of injected plasmid, it was possible to optimise the numbers of neurons that expressed the construct so that individual growth cones could be easily visualised. We then used time-lapse high magnification
widefield epifluorescence microscopy to visualise the growth cones as they were exploring their environment.  Growth cones both near the surface of the embryo as well as deep within the developing brain of embryos at 20 h post fertilisation were clearly imaged. With time-lapse sequence imaging with intervals between frames as frequent as 1 s there was minimal loss of fluorescence intensity and the dynamic nature of the growth cones became evident. This method therefore provides high
magnification, high resolution time-lapse imaging of living neurons in vivo and by use of widefield epifluorescence rather than confocal it is a relatively inexpensive microscopy method.
Keyword Axon
Time Lapse
Growth cone
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Biomedical Sciences Publications
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