Injection of Parhyale hawaiensis Blastomeres with Fluorescently Labeled Tracers
- Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3140, USA
- Department of Integrative Biology, University of California, Berkeley, CA 94720-3140, USA
- Howard Hughes Medical Institute, University of California, Berkeley, CA 94720-3140, USA
- ↵1Corresponding author (nipam{at}uclink.berkeley.edu)
INTRODUCTION
The great diversity of arthropod body plans, together with our detailed understanding of fruit fly development, makes arthropods a premier taxon for examining the evolutionary diversification of developmental patterns and hence the diversity of extant life. Crustaceans, in particular, show a remarkable range of morphologies and provide a useful outgroup to the insects. The amphipod crustacean Parhyale hawaiensis is becoming established as a model organism for developmental studies within the arthropods. This protocol describes the injection of P. hawaiensis blastomeres with fluorescently labeled tracers for the purpose of cell-lineage analysis. The total (holoblastic) cleavages that characterize early embryogenesis in P. hawaiensis generate an eight-cell embryo with a stereotypical arrangement of blastomeres, each of which already possesses an invariant cell fate. Fluorochrome-conjugated dextran solutions, mRNAs encoding fluorescent proteins, and biotin-dextran have all proven to be useful lineage markers. The relative merits of various tracers are considered.
