Epitope-tagging Math5 and Pou4f2: New tools to study retinal ganglion cell development in the mouse

Authors

  • Xueyao Fu,

    1. Department of Biochemistry and Molecular Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas
    Search for more papers by this author
    • Xueyao Fu and Takae Kiyama contributed equally to this article.

  • Takae Kiyama,

    1. Department of Biochemistry and Molecular Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas
    Search for more papers by this author
    • Xueyao Fu and Takae Kiyama contributed equally to this article.

  • Renzhong Li,

    1. Department of Ophthalmology/Ross Eye Institute and NYS Center of Excellence in Bioinformatics & Life Sciences, State University of New York at Buffalo, Buffalo, New York
    Search for more papers by this author
  • Mark Russell,

    1. Department of Biochemistry and Molecular Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas
    Search for more papers by this author
  • William H. Klein,

    1. Department of Biochemistry and Molecular Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas
    2. Graduate School of Biological Sciences, The University of Texas Health Science Center, Houston, Texas
    Search for more papers by this author
  • Xiuqian Mu

    Corresponding author
    1. Department of Biochemistry and Molecular Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas
    2. Department of Ophthalmology/Ross Eye Institute and NYS Center of Excellence in Bioinformatics & Life Sciences, State University of New York at Buffalo, Buffalo, New York
    • 701 Ellicott Street, Buffalo, New York 14203
    Search for more papers by this author

Abstract

Although immunological detection of proteins is used extensively in retinal development, studies are often impeded because antibodies against crucial proteins cannot be generated or are not readily available. Here, we overcome these limitations by constructing genetically engineered alleles for Math5 and Pou4f2, two genes required for retinal ganglion cell (RGC) development. Sequences encoding a peptide epitope from haemagglutinin (HA) were added to Math5 or Pou4f2 in frame to generate Math5HA and Pou4f2HA alleles. We demonstrate that the tagged alleles recapitulated the wild-type expression patterns of the two genes, and that the tags did not interfere with the function of the cognate proteins. In addition, by co-staining, we found that Math5 and Pou4f2 were transiently co-expressed in newly born RGCs, unequivocally demonstrating that Pou4f2 is immediately downstream of Math5 in RGC formation. The epitope-tagged alleles provide new and useful tools for analyzing gene regulatory networks underlying RGC development. Developmental Dynamics 238:2309–2317, 2009. © 2009 Wiley-Liss, Inc.

Ancillary