April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Establishment of A Mouse Cell Line using A Photoreceptor Gene-Specific Promoter and SV40 Large T antigen
Author Affiliations & Notes
  • Katsuhiro Hosono
    Ophthalmologly,
    Photon Medical Research Center,
    Hamamatsu University School of Medicine, Hamamatsu, Japan
  • Kentaro Ohishi
    Photon Medical Research Center,
    Hamamatsu University School of Medicine, Hamamatsu, Japan
  • Yoshitaka Yamaguchi
    Advanced Research Center for GSP, Keio University, Ibaraki, Japan
  • Jun Kudoh
    Advanced Research Center for GSP, Keio University, Ibaraki, Japan
    Laboratory of Gene Medicine, Keio University School of Medicine, Tokyo, Japan
  • Nobuyoshi Shimizu
    Advanced Research Center for GSP, Keio University, Ibaraki, Japan
  • Yoshihiro Hotta
    Ophthalmologly,
    Hamamatsu University School of Medicine, Hamamatsu, Japan
  • Shinsei Minoshima
    Photon Medical Research Center,
    Hamamatsu University School of Medicine, Hamamatsu, Japan
  • Footnotes
    Commercial Relationships  Katsuhiro Hosono, None; Kentaro Ohishi, None; Yoshitaka Yamaguchi, None; Jun Kudoh, None; Nobuyoshi Shimizu, None; Yoshihiro Hotta, None; Shinsei Minoshima, None
  • Footnotes
    Support  Grant-in-Aid for Scientific Research(B), Health Labour Science Research Grant
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 2253. doi:https://doi.org/
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      Katsuhiro Hosono, Kentaro Ohishi, Yoshitaka Yamaguchi, Jun Kudoh, Nobuyoshi Shimizu, Yoshihiro Hotta, Shinsei Minoshima; Establishment of A Mouse Cell Line using A Photoreceptor Gene-Specific Promoter and SV40 Large T antigen. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2253. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : We aimed at establishing mouse permanent cell lines that maintain characteristics of photoreceptor cells in order to use for studying the gene regulation on the onset of relevant eye diseases.

Methods: : We attempted to make a transgenic mouse (TgM) by using the unique transgene construct consisting of three genetic elements including the promoter sequence of cone-specific transducin alpha gene GNAT2, the coding sequence of SV40 Large T antigen (LT) and the enhancer sequence of human RBP3 which is effective on the gene expression in rod and cones of eye.

Results: : The resulting TgMs developed brain tumors, which were believed to be of pineal gland origin. The TgMs also showed apoptosis-like degeneration in the photoreceptor cells. The tumor tissue was dispersed and the dissociated cells were kept in culture with periodical transfer for 240 days. After cloning by limiting dilution, a cell line named G2P-7 with a doubling time of 26.6 h was established. Analysis of gene expression by RT-PCR and DNA microarray revealed that G2P-7 cells express cone-specific Gnat2 but not rod-specific Gnat1. Furthermore, a luciferase reporter assay using the promoters of relevant genes were performed. The promoters of Gnat2 (cone-specific), Rbp3 (rod as well as cone-specific) and Tph1 (pineal gland-specific) showed significant promoter activity but Gnat1 (rod-specific) did not.

Conclusions: : We established a mouse cell line, G2P-7, that shows the expression of cone-specific Gnat2 but not rod-specific Gnat1. It is considered that G2P-7 might maintain some characteristics of cone cells. This cell line would be useful for studying the gene regulation occurring in the photoreceptor cells and pineal gland.

Keywords: gene/expression • retinal culture • photoreceptors 
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