The Fugu tyrp1 promoter directs specific GFP expression in zebrafish: tools to study the RPE and the neural crest-derived melanophores

Authors

  • Jian Zou,

    1. Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA
    Search for more papers by this author
  • Friedrich Beermann,

    1. ISREC (Swiss Institute for Experimental Cancer Research), National Center of Competence in Research (NCCR) Molecular Oncology, Chemin des Boveresses, 1066 Epalinges, Switzerland
    Search for more papers by this author
  • Jianxin Wang,

    1. Znomics, Inc. 2611 S.W. 3rd Ave., Suite 200, Portland, OR 97201, USA
    Search for more papers by this author
  • Koichi Kawakami,

    1. Division of Molecular and Developmental Biology, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-8540, Japan
    Search for more papers by this author
  • Xiangyun Wei

    Corresponding author
    1. Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA
    2. Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA
    Search for more papers by this author

*Address correspondence to Xiangyun Wei, e-mail: weix@upmc.edu

Summary

In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish.

Ancillary