Injection of Parhyale hawaiensis blastomeres with fluorescently labeled tracers.
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.
Rehm
E Jay
EJ
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3140, USA.
Hannibal
Roberta L
RL
Chaw
R Crystal
RC
Vargas-Vila
Mario A
MA
Patel
Nipam H
NH
D033304:Amphipoda / Q000196:embryology*; D000818:Animals; D001710:Biotin / Q000737:chemistry; D001757:Blastomeres / Q000166:cytology*; D019070:Cell Lineage; D015509:Developmental Biology / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism
D003911:Dextrans; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D001710:Biotin