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Methodology for Whole Mount and Fluorescent RNA In Situ Hybridization in Echinoderms: Single, Double, and Beyond

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Developmental Biology of the Sea Urchin and Other Marine Invertebrates

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2219))

Abstract

Identifying the location of a specific RNA in a cell, tissue, or embryo is essential to understand its function. Here we use echinoderm embryos to demonstrate the power of fluorescence in situ RNA hybridizations to localize sites of specific RNA accumulation in whole mount embryo applications. We add to this technology the use of various probe-labeling technologies to colabel multiple RNAs in one application and we describe protocols for incorporating immunofluorescence approaches to maximize the information obtained in situ. We offer alternatives for these protocols and troubleshooting advice to identify steps in which the procedure may have failed. Overall, echinoderms are wonderfully suited for these technologies, and these protocols are applicable to a wide range of cells, tissues, and embryos.

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Author information

Authors and Affiliations

  1. Department of Molecular and Cellular Biology and Biochemistry, Brown University, Providence, RI, USA

    Margherita Perillo, Maxwell Spurrell & Gary M. Wessel

  2. Department of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Naples, Italy

    Periklis Paganos & Maria I. Arnone

Authors
  1. Margherita Perillo
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  2. Periklis Paganos
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  3. Maxwell Spurrell
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  4. Maria I. Arnone
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  5. Gary M. Wessel
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Corresponding authors

Correspondence to Maria I. Arnone or Gary M. Wessel .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, Midwestern University, Glendale, AZ, USA

    David J. Carroll

  2. Department of Biology, University of New Mexico, Albuquerque, NM, USA

    Stephen A. Stricker

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Perillo, M., Paganos, P., Spurrell, M., Arnone, M.I., Wessel, G.M. (2021). Methodology for Whole Mount and Fluorescent RNA In Situ Hybridization in Echinoderms: Single, Double, and Beyond. In: Carroll, D.J., Stricker, S.A. (eds) Developmental Biology of the Sea Urchin and Other Marine Invertebrates. Methods in Molecular Biology, vol 2219. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0974-3_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0974-3_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0973-6

  • Online ISBN: 978-1-0716-0974-3

  • eBook Packages: Springer Protocols

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