May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Distal conserved region is required for the rod photoreceptor specific expression of Nrl in both mouse and frog
M. Akimoto; L. Whitaker; J.F. Friedman; H. Khanna; E. Filippova; A.J. Mears; D.L. Turner; B.E. Knox; A. Swaroop
Author Affiliations & Notes
  • M. Akimoto
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
    Translational Research Center, Kyoto University, Kyoto, Japan
  • L. Whitaker
    Biochemistry & Molecular Biology and Ophthalmology, SUNY Upstate Medical University, Syracuse, NY
  • J.F. Friedman
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
  • H. Khanna
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
  • E. Filippova
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
  • A.J. Mears
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
    Ottawa Health Research Institute, Ottawa, ON, Canada
  • D.L. Turner
    Mental Health Research Institute,
    University of Michigan, Ann Arbor, MI
  • B.E. Knox
    Biochemistry & Molecular Biology and Ophthalmology, SUNY Upstate Medical University, Syracuse, NY
  • A. Swaroop
    Ophthalmology and Human Genetics, WK Kellogg Eye Center,
    University of Michigan, Ann Arbor, MI
  • Footnotes
    Commercial Relationships  M. Akimoto, None; L. Whitaker, None; J.F. Friedman, None; H. Khanna, None; E. Filippova, None; A.J. Mears, None; D.L. Turner, None; B.E. Knox, None; A. Swaroop, None.
  • Footnotes
    Support  NIH–EY11115, FFB and RPB
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 2251. doi:
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      M. Akimoto, L. Whitaker, J.F. Friedman, H. Khanna, E. Filippova, A.J. Mears, D.L. Turner, B.E. Knox, A. Swaroop; Distal conserved region is required for the rod photoreceptor specific expression of Nrl in both mouse and frog . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2251.

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Abstract

Abstract: : Purpose: NRL is required for the expression of rod–specific genes and is essential for rod photoreceptor differentiation. The goal of this study is to identify cis–regulatory elements and trans–acting factors that control the rod–specific expression of NRL in developing retina. Methods: We studied the regulation of the Nrl gene using transgenic mice with three different lengths of 5’ –flanking promoter sequences of mouse Nrl tagged to the GFP reporter gene. Similar constructs with human NRL promoter sequences were used for studies in transgenic Xenopus. DNaseI footprinting was carried out for all conserved sequences within the Nrl–5’–upstream region. Gelshift and supershift assays were performed using bovine retinal nuclear extracts. Co–transfection of candidate regulators, myocyte enhancer factor (Mef2C) and bHLH proteins, were performed in cultured cells using luciferase reporter assays. Results: Four highly conserved regions (I–IV) were identified between the 5’–upstream regions of the human and mouse Nrl genes. The distal conserved region IV contains Mef2 and E–box–like sites. Two different deletion mutant promoters of this region result in ectopic expression of GFP in both mouse and frog. Supershift assay revealed specific binding of Mef2C to the cis–element. Mef2C and Mash1 cooperatively transactivate Nrl expression, whereas regulation by NeuroD is mediated by the proximal conserved region I. Conclusions: Distal conserved region of the Nrl promoter is required for rod –specific expression. Mef2C and bHLH proteins play an important role in Nrl expression in early rod progenitors.

Keywords: retinal development • transcription factors • transgenics/knock–outs 
© 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 © 2025 Association for Research in Vision and Ophthalmology.
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