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Imaging and Quantification of P-Smad1/5 in Zebrafish Blastula and Gastrula Embryos

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Bone Morphogenetic Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1891))

Abstract

Spatiotemporal patterns of morphogen activity drive differential gene expression with a high degree of precision within a developing embryo and reproducibly between embryos. Understanding the formation and function of a morphogen gradient during development requires quantitative measurement of morphogen activity throughout an individual embryo and also between embryos within a population. Quantification of morphogen gradients in to presents unique challenges in imaging and image processing to minimize error and maximize the quality of the data so it may be used in computational models of development and in statistically testing hypotheses. Here we present methods for the preparation, immunostaining, imaging, and quantification of a bone morphogenetic protein (BMP) activity gradient in individual zebrafish embryos as well as methods for acquiring population-level statistics after embryo grouping and alignment. This quantitative approach can be extended to other morphogen systems, and the computational codes can be adapted to other imaging contexts in zebrafish and other organisms.

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Acknowledgment

This work was supported by grant NIH R01GM056326 and NIH R01HD073156.

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Joseph Zinski, Francesca Tuazon & Mary Mullins

  2. Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN, USA

    Yan Huang & David Umulis

  3. Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA

    David Umulis

Authors
  1. Joseph Zinski
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  2. Francesca Tuazon
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  3. Yan Huang
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  4. Mary Mullins
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  5. David Umulis
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Corresponding authors

Correspondence to Mary Mullins or David Umulis .

Editor information

Editors and Affiliations

  1. New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA

    Melissa B. Rogers

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Zinski, J., Tuazon, F., Huang, Y., Mullins, M., Umulis, D. (2019). Imaging and Quantification of P-Smad1/5 in Zebrafish Blastula and Gastrula Embryos. In: Rogers, M. (eds) Bone Morphogenetic Proteins. Methods in Molecular Biology, vol 1891. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8904-1_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8904-1_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8903-4

  • Online ISBN: 978-1-4939-8904-1

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