Cite as: Cold Spring Harb. Protoc.; 2008; doi:10.1101/pdb.prot5097

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Analysis of Cell Movement in Amphimedon Embryos by Injection of Fluorescent Tracers

Maja Adamska1,2 and Bernard M. Degnan1,3

1 School of Integrative Biology, University of Queensland, Brisbane QLD 4072, Australia
2 Sars International Centre for Marine Molecular Biology, N-5008 Bergen, Norway

3Corresponding author (b.degnan{at}uq.edu.au)


INTRODUCTION

Although attempts to culture prepigmentation-stage embryos (i.e., blastulas and early gastrulas) outside of brood chambers have so far been unsuccessful in Amphimedon, it is possible to manipulate embryos within the brood chamber and follow their development under laboratory conditions. This protocol describes microinjection of lipophilic tracers such as 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) into embryos embedded in their native brood chamber. DiI does not appear to perturb embryonic development and is relatively resistant to photobleaching. As long as care is taken not to damage the fragile embryos during observation and photography, the same embryo can be photographed multiple times, permitting its development to be tracked (up to 4 wk) from early cleavage stages to hatching of free-swimming parenchymella larvae. The embryos or larvae also can be fixed without loss of fluorescence. This method also can be used to deliver other types of solutions to embryos or individual cells of early embryos.


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Related Article

The Demosponge Amphimedon queenslandica: Reconstructing the Ancestral Metazoan Genome and Deciphering the Origin of Animal Multicellularity
Bernard M. Degnan, Maja Adamska, Alina Craigie, Sandie M. Degnan, Bryony Fahey, Marie Gauthier, John N.A. Hooper, Claire Larroux, Sally P. Leys, Erica Lovas, and Gemma S. Richards
Cold Spring Harb Protoc 2008: 108. [Abstract] [Full Text]

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Isolation of Amphimedon Developmental Material
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Home page
 Cold Spring Harb ProtocHome page
B. M. Degnan, M. Adamska, A. Craigie, S. M. Degnan, B. Fahey, M. Gauthier, J. N.A. Hooper, C. Larroux, S. P. Leys, E. Lovas, et al.
The Demosponge Amphimedon queenslandica: Reconstructing the Ancestral Metazoan Genome and Deciphering the Origin of Animal Multicellularity
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.emo108 - pdb.emo108.
[Abstract] [Full Text]