The basic helix-loop-helix transcription factor, heart and neural crest derivatives expressed transcript 2, marks hepatic stellate cells in zebrafish: Analysis of stellate cell entry into the developing liver§

Authors

  • Chunyue Yin,

    1. Department of Biochemistry and Biophysics, Programs in Developmental and Stem Cell Biology, Genetics and Human Genetics, Liver Center and Diabetes Center, Institute for Regeneration Medicine, University of California San Francisco, San Francisco, CA
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    • *These authors contribute equally to this work.

  • Kimberley J. Evason,

    1. Department of Biochemistry and Biophysics, Programs in Developmental and Stem Cell Biology, Genetics and Human Genetics, Liver Center and Diabetes Center, Institute for Regeneration Medicine, University of California San Francisco, San Francisco, CA
    2. Department of Pathology, University of California San Francisco, San Francisco, CA
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    • *These authors contribute equally to this work.

  • Jacquelyn J. Maher,

    1. Department of Medicine, and Liver Center, University of California San Francisco, San Francisco, CA
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  • Didier Y.R. Stainier

    Corresponding author
    1. Department of Biochemistry and Biophysics, Programs in Developmental and Stem Cell Biology, Genetics and Human Genetics, Liver Center and Diabetes Center, Institute for Regeneration Medicine, University of California San Francisco, San Francisco, CA
    • Department of Biochemistry and Biophysics, Programs in Developmental and Stem Cell Biology, Genetics and Human Genetics, Liver Center and Diabetes Center, Institute for Regeneration Medicine, University of California, San Francisco, 1550 Fourth Street, Room 384, San Francisco, CA 94158
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    • fax: 415-476-3892


  • See Editorial on Page 1596

  • Potential conflict of interest: Nothing to report.

  • §

    C.Y. is supported by grant no. K99AA020514 from the National Institutes of Health (NIH) and the University of California San Francisco Liver Center Pilot/Feasibility Award (NIH P30DK026743). K.J.E. is a Robert Black Fellow of the Damon Runyon Cancer Research Foundation (DRG-109-10). J.J.M. is supported by grants from the NIH (R01DK068450, R01DK088674, and P30DK026743). This work was supported, in part, by grants from the NIH (R01DK060322) and the Packard Foundation (to D.Y.R.S.).

Abstract

Hepatic stellate cells (HSCs) are liver-specific mesenchymal cells that play vital roles in liver development and injury. Our knowledge of HSC biology is limited by the paucity of in vivo data. HSCs and sinusoidal endothelial cells (SECs) reside in close proximity, and interactions between these two cell types are potentially critical for their development and function. Here, we introduce a transgenic zebrafish line, Tg(hand2:EGFP), that labels HSCs. We find that zebrafish HSCs share many similarities with their mammalian counterparts, including morphology, location, lipid storage, gene-expression profile, and increased proliferation and matrix production, in response to an acute hepatic insult. Using the Tg(hand2:EGFP) line, we conducted time-course analyses during development to reveal that HSCs invade the liver after SECs do. However, HSCs still enter the liver in mutants that lack most endothelial cells, including SECs, indicating that SECs are not required for HSC differentiation or their entry into the liver. In the absence of SECs, HSCs become abnormally associated with hepatic biliary cells, suggesting that SECs influence HSC localization during liver development. We analyzed factors that regulate HSC development and show that inhibition of vascular endothelial growth factor signaling significantly reduces the number of HSCs that enter the liver. We also performed a pilot chemical screen and identified two compounds that affect HSC numbers during development. Conclusion: Our work provides the first comprehensive description of HSC development in zebrafish and reveals the requirement of SECs in HSC localization. The Tg(hand2:EGFP) line represents a unique tool for in vivo analysis and molecular dissection of HSC behavior. (HEPATOLOGY 2012;56:1958–1970)

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