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Antibody Uptake Assay in the Embryonic Zebrafish Forebrain to Study Notch Signaling Dynamics in Neural Progenitor Cells In Vivo

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Organoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1576))

Abstract

Stem cells can generate cell fate heterogeneity through asymmetric cell division (ACD). ACD derives from the asymmetric segregation of fate-determining molecules and/or organelles in the dividing cell. Radial glia in the embryonic zebrafish forebrain are an excellent model for studying the molecular mechanisms regulating ACD of stem cells in vertebrates, especially for live imaging concerning in vivo molecular and cellular dynamics. Due to the current difficulty in expressing fluorescent reporter-tagged proteins at physiological levels in zebrafish for live imaging, we have developed an antibody uptake assay to label proteins in live embryonic zebrafish forebrain with high specificity. DeltaD is a transmembrane ligand in Notch signaling pathway in the context of ACD of radial glia in zebrafish. By using this assay, we have successfully observed the in vivo dynamics of DeltaD for studying ACD of radial glia in the embryonic zebrafish forebrain.

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Acknowledgments

This work was supported by NIH (R01 NS095734) and Fudan Bio-elite program. We thank Xiang Zhao for discussions on the antibody uptake assay and comments on the manuscript, as well as Maximilian Fürthauer for sharing experience on the antibody uptake assay in the zebrafish spinal cord.

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Authors and Affiliations

  1. State Key Laboratory of Genetic Engineering, Department of Genetics, School of Life Sciences, Fudan University, Shanghai, China

    Kai Tong & Su Guo

  2. Department of Bioengineering and Therapeutic Sciences, Programs in Human Genetics and Biological Sciences, ELi and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, CA, USA

    Kai Tong, Mahendra Wagle & Su Guo

Authors
  1. Kai Tong
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  2. Mahendra Wagle
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  3. Su Guo
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Corresponding author

Correspondence to Su Guo .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, ON, Canada

    Kursad Turksen

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Tong, K., Wagle, M., Guo, S. (2017). Antibody Uptake Assay in the Embryonic Zebrafish Forebrain to Study Notch Signaling Dynamics in Neural Progenitor Cells In Vivo. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2017_22

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  • DOI: https://doi.org/10.1007/7651_2017_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-7616-4

  • Online ISBN: 978-1-4939-7617-1

  • eBook Packages: Springer Protocols

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