May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Co–Localization of EFEMP1 and TIMP3 in the Developing and Mature Mouse Retina
Author Affiliations & Notes
  • D.C. Chung
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • N.W. Keiser
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • J.L. Bennicelli
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • W. Tang
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • Z. Wei
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • J. Bennett
    FM Kirby Ctr for Molecular Ophth, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA
  • Footnotes
    Commercial Relationships  D.C. Chung, None; N.W. Keiser, None; J.L. Bennicelli, None; W. Tang, None; Z. Wei, None; J. Bennett, None.
  • Footnotes
    Support  : RO1 EY12156, T32DK07748, Foundation Fighting Blindness, Lew Wasserman Research to Prevent Blindn
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 1637. doi:
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      D.C. Chung, N.W. Keiser, J.L. Bennicelli, W. Tang, Z. Wei, J. Bennett; Co–Localization of EFEMP1 and TIMP3 in the Developing and Mature Mouse Retina . Invest. Ophthalmol. Vis. Sci. 2005;46(13):1637.

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

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Abstract

Abstract: : Purpose:To evaluate the immunofluorescent localization patterns of EFEMP1 and TIMP3 in the developing and mature mouse retina. Methods: Eyes of CD1 mice at developmental stages E14, P2, P7, P12, P14, and 4 months were collected after euthanasia. Eyes were fixed in 4% paraformaldehyde, cryoprotected and sectioned at 10µm. Sections were blocked, incubated with a polyclonal rabbit anti–EFEMP1 antibody generated in our lab and with a mouse anti–human TIMP–3 antibody (Calbiochem, San Diego, CA). After washing, sections were incubated in secondary antibodies anti–rabbit IgG Alexa 488 and anti–mouse IgG Alexa 594 (Molecular Probes, Eugene, OR). The sections were washed and stained with DAPI and fluorescence was evaluated on a Leica fluorescent microscope. Results: EFEMP1 and TIMP3 co–localized in RPE at E14, although EFEMP1 was in the apical portion of the cells and TIMP3 was basal and in Bruch’s membrane. Co–localization was also found in the nerve fiber layer. At P2 and P7, there was increased co–localization in the nerve fiber layer, retinal ganglion cells, and the choroid, but not RPE. At P12 and older, co–localization persisted in the retinal ganglion cell layer. EFEMP1, however, then appeared in the basal portion of RPE and the inner segments of the photoreceptors. Conclusions: EFEMP1 and TIMP–3 co–localize to the inner retina, and depending on development stage, in the RPE. The interaction of these two proteins may play a role in angiogenesis in the retina.

Keywords: retinal degenerations: cell biology • extracellular matrix • retinal development 
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