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Lymphangiogenesis pp 55–70Cite as

Visualization and Tools for Analysis of Zebrafish Lymphatic Development

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1846))

Abstract

The accessibility and optical transparency of the zebrafish embryo offers a unique platform for live-imaging of developmental lymphangiogenesis. Transgenic lines labelling lymphatic progenitors and vessels enable researchers to visualize cellular processes and ask how they contribute to lymphatic development in genetic models. Furthermore, validated immunofluorescence staining for key signaling and cell fate markers (phosphorylated Erk and Prox1) allow single cell resolution studies of lymphatic differentiation. Here, we describe in detail how zebrafish embryos and larvae can be mounted for high resolution, staged imaging of lymphatic networks, how lymphangiogenesis can be reliably quantified and how immunofluorescence can reveal lymphatic signaling and differentiation. These methods offer researchers the opportunity to experimentally dissect developmental lymphangiogenesis with outstanding resolution.

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Acknowledgments

We would like to thank Ms. Tevin Chau and Dr. Neil Bower for their technical assistance. The IF protocol for phosphorylated Erk and Prox1 is our own modified version of protocols originally carefully optimized and published by Professor Nathan Lawson and Dr. Masahiro Shin from the University of Massachusetts Medical School [32, 34] and by Professor Joachim Wittbrodt and Dr. Daigo Inoue from the Heidelberg University [35].

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Author notes

  1. Sungmin Baek

    Present address: Stowers Institute for Medical Research, Kansas city, MO, USA

Authors and Affiliations

  1. Division of Genomics of Development and Disease, Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia

    Kazuhide S. Okuda, Sungmin Baek & Benjamin M. Hogan

Authors
  1. Kazuhide S. Okuda
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  2. Sungmin Baek
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  3. Benjamin M. Hogan
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Corresponding author

Correspondence to Benjamin M. Hogan .

Editor information

Editors and Affiliations

  1. Center for Vascular & Developmental Biology, Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, IL, USA

    Guillermo Oliver

  2. Department of Medicine and Cardiovascular Institute, University of Pennsylvania, Philadelphia, PA, USA

    Mark L. Kahn

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Okuda, K.S., Baek, S., Hogan, B.M. (2018). Visualization and Tools for Analysis of Zebrafish Lymphatic Development. In: Oliver, G., Kahn, M. (eds) Lymphangiogenesis. Methods in Molecular Biology, vol 1846. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8712-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-8712-2_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8711-5

  • Online ISBN: 978-1-4939-8712-2

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