Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5128

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Injection of Parhyale hawaiensis Blastomeres with Fluorescently Labeled Tracers

E. Jay Rehm, Roberta L. Hannibal, R. Crystal Chaw, Mario A. Vargas-Vila, and Nipam H. Patel1

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)

This article is also available in Emerging Model Organisms: A Laboratory Manual, Vol. 1. CSHL Press, Cold Spring Harbor, NY, USA, 2009.


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.


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The Crustacean Parhyale hawaiensis: A New Model for Arthropod Development
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E. J. Rehm, R. L. Hannibal, R. C. Chaw, M. A. Vargas-Vila, and N. H. Patel
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