May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Vascular Endothelial Growth Factor Over-Expression in Transgenic Zebrafish Retina
C.-Y. Hu; C.-H. Yang; H.-Y. Huang; C.-T. Tu; H.-J. Tsai
Author Affiliations & Notes
  • C.-Y. Hu
    Department of Ophthalmology, Far Eastern Memorial Hospital, Taipei, Taiwan
  • C.-H. Yang
    Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
  • H.-Y. Huang
    Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan
  • C.-T. Tu
    Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan
  • H.-J. Tsai
    Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan
  • Footnotes
    Commercial Relationships C. Hu, None; C. Yang, None; H. Huang, None; C. Tu, None; H. Tsai, None.
  • Footnotes
    Support National Science Council Grant 94-2314-B-418-010, Taiwan.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 3421. doi:
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      C.-Y. Hu, C.-H. Yang, H.-Y. Huang, C.-T. Tu, H.-J. Tsai; Vascular Endothelial Growth Factor Over-Expression in Transgenic Zebrafish Retina. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3421.

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Abstract

Purpose:: We intended to establish a zebrafish model of vascular endothelial growth factor (VEGF) over-expression in retina, so that it may become a useful animal model of retinal or choroidal neovascularization.

Methods:: We micro-injected a construct of zebrafish VEGF driven by retina-specific rhodopsin promoter and an additional construct of red fluorescent protein (RFP) driven by heart-specific cmlc2 promoter into the embryos derived from zebrafish transgenic line Tg(fli1:EGFP), which highly expresses enhanced green fluorescent protein (EGFP) in vascular endothelium. We cultured the larvae with heart-specific RFP expression and mated them with each other after sexual maturation. We used polymerase chain reaction (PCR) to identify the existence of rhodopsin promoter and VEGF coding sequence in the genome of F1 progeny to prove successful inheritance of transgene. Semi-quantitative reverse transcription PCR (RT-PCR) was applied to compare the expression levels of VEGF between the Tg(fli1:EGFP) larvae and the Tg(rhodopsin:VEGF) larvae. The alterations in ocular tissue were confirmed by histological examination.

Results:: Hundreds of embryos from line Tg(fli1:EGFP) were micro-injected with the linearized plasmid DNA. The putative founders expressing RFP in heart were cultured for 3 to 4 months. PCR analysis confirmed the stable existence of transgene by identifying the injected sequence in the genomic DNA of F1 larvae. Semi-quantitative RT-PCR also demonstrated that the VEGF expression level of Tg(rhodopsin:VEGF) larvae was higher than that of Tg(fli1:EGFP) larvae. Histological examination of the PCR-confirmed F1 adult fish showed increased number and size of blood vessels in choroid and vitreous cavity, but the sensory retina did not change apparently.

Conclusions:: We successfully established a zebrafish model of VEGF over-expression in retina. Its long-term influence on retinal and choroidal vasculogenesis deserves further more detailed investigation.

Keywords: choroid: neovascularization • retinal neovascularization 
© 2007, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2024 Association for Research in Vision and Ophthalmology.
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