May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
The change in the early retinal development of zebrafish after Egr1 gene knockdown
C.–Y. Hu; C.–H. Yang; W.–Y. Chen; Y.–H. Chen; C.–J. Huang; H.–J. Tsai
Author Affiliations & Notes
  • C.–Y. Hu
    Department of Ophthalmology, Far Eastern Memorial Hospital, Taipei, Taiwan Republic of China
  • C.–H. Yang
    Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan Republic of China
  • W.–Y. Chen
    Department of Pathology, Taipei Medical University, Taipei, Taiwan Republic of China
  • Y.–H. Chen
    Institute of Molecular and Cell Biology, National Taiwan University, Taipei, Taiwan Republic of China
  • C.–J. Huang
    Institute of Molecular and Cell Biology, National Taiwan University, Taipei, Taiwan Republic of China
  • H.–J. Tsai
    Institute of Molecular and Cell Biology, National Taiwan University, Taipei, Taiwan Republic of China
  • Footnotes
    Commercial Relationships  C. Hu, None; C. Yang, None; W. Chen, None; Y. Chen, None; C. Huang, None; H. Tsai, None.
  • Footnotes
    Support  NSC Grant 92–2314–B–418–006
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 687. doi:
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      C.–Y. Hu, C.–H. Yang, W.–Y. Chen, Y.–H. Chen, C.–J. Huang, H.–J. Tsai; The change in the early retinal development of zebrafish after Egr1 gene knockdown . Invest. Ophthalmol. Vis. Sci. 2004;45(13):687.

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Abstract

Abstract: : Purpose: Egr1 (early growth response 1) gene has been proposed to be associated with abnormal eye growth. Our aim is to identify the change in early retinal development after Egr1 gene knockdown. Methods: We microinjected the anti–sense mRNA inhibitor, Egr1–morpholino (Gene Tools), into the 1– to 4–cell zebrafish embryos. Chordin morpholino and the standard control oligo were also injected separately as positive and negative controls. The expression rate was calculated according to their morphologic changes. At 72 hours post–fertilization (hpf), larvae were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned into 5 µm slices, stained and observed under light microscope. Immunohistochemical staining was applied to identify amacrine cells. Results: Hundreds of zebrafish embryos have been microinjected. Seven different dosage of Egr1–morpholino was tried, and the most appropriate dose according to its efficacy and lethality at 24–hpf was 13.5 ng per embryo. More than 70% of survival embryos at 48–hpf had altered phenotype, yet only <1% of live negative control did. The most common defects of Egr1 morphants were small eyes as well as enlarged pericardium. Western blot analysis showed the absence of 55–kD Egr1 protein in morphants. Histological examination of 72–hpf morphants revealed smaller eyes with a decreased cell number and a thinner inner plexiform layer. Immunohistochemical study suggested a decrease of amacrine cells. Conclusions: Egr1 gene plays an important role in the early retinal development, especially for the amacrine cells. However, the mechanism of abnormal eye growth is still to be determined.

Keywords: retinal development • amacrine cells 
© 2004, 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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