Skip to main content
SpringerLink
Log in

Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids

  • Protocol
  • First Online:
The Hippo Pathway

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1893))

Abstract

The transcription coactivator, Yes-associated protein (YAP), which is a nuclear effector of the Hippo signaling pathway, has been shown to be a mechano-transducer. By using mutant fish and human 3D spheroids, we have recently demonstrated that YAP is also a mechano-effector. YAP functions in three-dimensional (3D) morphogenesis of organ and global body shape by controlling actomyosin-mediated tissue tension. In this chapter, we present a platform that links the findings in fish embryos with human cells. The protocols for analyzing tissue tension-mediated global body shape/organ morphogenesis in vivo and ex vivo using medaka fish embryos and in vitro using human cell spheroids represent useful tools for unraveling the molecular mechanisms by which YAP functions in regulating global body/organ morphogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Porazinski S, Wang H, Asaoka Y, Behrndt M, Miyamoto T, Morita H et al (2015) YAP is essential for tissue tension to ensure vertebrate 3D body shape. Nature 521:217–221

    Article  CAS  Google Scholar 

  2. Furutani-Seiki M, Sasado T, Morinaga C, Suwa H, Niwa K, Yoda H et al (2004) A systematic genome-wide screen for mutations affecting organogenesis in Medaka, Oryzias latipes. Mech Dev 121:647–658

    Google Scholar 

  3. Furutani-Seiki M, Wittbrodt J (2004) Medaka and zebrafish, an evolutionary twin study. Mech Dev 121:629–637

    Article  CAS  Google Scholar 

  4. Watanabe T, Asaka S, Kitagawa D, Saito K, Kurashige R, Sasado T et al (2004) Mutations affecting liver development and function in Medaka, Oryzias latipes, screened by multiple criteria. Mech Dev 121:791–802

    Article  CAS  Google Scholar 

  5. Nelson CM, Bissell MJ (2006) Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Annu Rev Cell Dev Biol 22:287–309

    Article  CAS  Google Scholar 

  6. Humphrey JD, Dufresne ER, Schwartz MA (2014) Mechanotransduction and extracellular matrix homeostasis. Nat Rev Mol Cell Biol 15:802–812

    Article  CAS  Google Scholar 

  7. Halder G, Dupont S, Piccolo S (2012) Transduction of mechanical and cytoskeletal cues by YAP and TAZ. Nat Rev Mol Cell Biol 13:591–600

    Article  CAS  Google Scholar 

  8. Calvo F, Ege N, Grande-Garcia A, Hooper S, Jenkins RP, Chaudhry SI et al (2013) Mechanotransduction and YAP-dependent matrix remodelling is required for the generation and maintenance of cancer-associated fibroblasts. Nat Cell Biol 15:637–646

    Article  CAS  Google Scholar 

  9. Butcher DT, Alliston T, Weaver VM (2009) A tense situation: forcing tumour progression. Nat Rev Cancer 9:108–122

    Article  CAS  Google Scholar 

  10. Antoni D, Burckel H, Josset E, Noel G (2015) Three-dimensional cell culture: a breakthrough in vivo. Int J Mol Sci 16:5517–5527

    Article  CAS  Google Scholar 

  11. Heisenberg CP, Bellaïche Y (2013) Forces in tissue morphogenesis and patterning. Cell 153:948–962

    Article  CAS  Google Scholar 

  12. Porazinski SR, Wang H, Furutani-Seiki M (2011) Essential techniques for introducing medaka to a zebrafish laboratory—towards the combined use of medaka and zebrafish for further genetic dissection of the function of the vertebrate genome. Methods Mol Biol 770:211–241

    Article  CAS  Google Scholar 

  13. Maître JL, Berthoumieux H, Krens SF, Salbreux G, Jülicher F, Paluch E et al (2012) Adhesion functions in cell sorting by mechanically coupling the cortices of adhering cells. Science 338:253–256

    Article  Google Scholar 

Download references

Acknowledgments

We would like to thank Hiroshi Nishina (Tokyo Medical and Dental University, Japan), Hisato Kondoh (Kyoto Sangyo University, Japan), and other collaborators for the Kyoto medaka mutant screening. The research of gravity and YAP was partially supported by JSPS KAKENHI Grant Numbers 16H01643, 16H01644, and 17H05769. Yoichi Asaoka and Hitoshi Morita contributed equally to this chapter.

Author information

Author notes

  1. Yoichi Asaoka and Hitoshi Morita contributed equally to this work.

Authors and Affiliations

  1. Department of Systems Biochemistry in Pathology and Regeneration, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan

    Yoichi Asaoka

  2. Laboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Sciences, University of Yamanashi, Yamanashi, Japan

    Hitoshi Morita

  3. IST Austria, Klosterneuburg, Austria

    Carl-Philipp Heisenberg

  4. Department of Systems Biochemistry in Pathology and Regeneration, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan

    Hiroko Furumoto & Makoto Furutani-Seiki

Authors
  1. Yoichi Asaoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hitoshi Morita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroko Furumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Carl-Philipp Heisenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Makoto Furutani-Seiki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carl-Philipp Heisenberg .

Editor information

Editors and Affiliations

  1. Cancer Institute, University College London, London, UK

    Alexander Hergovich

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Asaoka, Y., Morita, H., Furumoto, H., Heisenberg, CP., Furutani-Seiki, M. (2019). Studying YAP-Mediated 3D Morphogenesis Using Fish Embryos and Human Spheroids. In: Hergovich, A. (eds) The Hippo Pathway. Methods in Molecular Biology, vol 1893. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8910-2_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8910-2_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8909-6

  • Online ISBN: 978-1-4939-8910-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation