May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Retinal Ganglion Cell Evaluation Using Thy1–Cyan Fluorescent Protein Mice in Optic Nerve Crush Model
F. Mabuchi; M. Aihara; J.D. Lindsey; M.R. Mackey; J.G. Crowston; R.N. Weinreb
Author Affiliations & Notes
  • F. Mabuchi
    Ophthalmology, University of Yamanashi, Tamaho, Japan
    Hamilton Glaucoma Center, University of California, San Diego, La Jolla, CA
  • M. Aihara
    Hamilton Glaucoma Center, University of California, San Diego, La Jolla, CA
    Ophthalmology, University of Tokyo, Hongo, Japan
  • J.D. Lindsey
    Hamilton Glaucoma Center, University of California, San Diego, La Jolla, CA
  • M.R. Mackey
    National Center for Microscopy and Imaging Research and Department of Neurosciences, University of California San Diego, La Jolla, CA
  • J.G. Crowston
    Hamilton Glaucoma Center, University of California, San Diego, La Jolla, CA
  • R.N. Weinreb
    Hamilton Glaucoma Center, University of California, San Diego, La Jolla, CA
  • Footnotes
    Commercial Relationships  F. Mabuchi, None; M. Aihara, None; J.D. Lindsey, None; M.R. Mackey, None; J.G. Crowston, None; R.N. Weinreb, None.
  • Footnotes
    Support  NIH Grant EY05990
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 5361. doi:
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      F. Mabuchi, M. Aihara, J.D. Lindsey, M.R. Mackey, J.G. Crowston, R.N. Weinreb; Retinal Ganglion Cell Evaluation Using Thy1–Cyan Fluorescent Protein Mice in Optic Nerve Crush Model . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5361.

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Abstract

Abstract: : Purpose: To evaluate retinal ganglion cell (RGC) changes following optic nerve crush in Thy1–Cyan fluorescent protein (CFP) transgenic mice. Methods: Nine Thy1–CFP mice, in which the Thy1 promoter drives expression of CFP, were anestheized and the optic nerve of one eye was crushed. Two or three weeks after surgery, the mice were transcardially fixed, the retinas were dissected, and flat mounts of the retinas were prepared. Images of the optic nerve axons and RGCs were systematically collected using fluorescence microscopy. These images were masked prior to assessment of RGC density. The survival of RGCs in the treated eyes was estimated and compared among the central, mid–peripheral, and peripheral regions of the retina. The morphology of individual RGCs was assessed by confocal microscopy. Individual images (1024X1024 pixels) as well as z–series stacks (90–170 images at 0.05 mm z–spacing) were collected and later merged to produce high–resolution three–dimensional reconstructions of individual RGCs. Results: Fluorescent RGCs and amacrine cells were readily distinguished by conventional fluorescence microscopy. Survival of RGCs in the treated eyes was 11.8 ± 1.6 % (mean ± SD, n = 5) at 2 weeks after surgery and 4.4 ± 0.5 % (n = 4) at 3 weeks after surgery. RGC survival in the central retina was significantly lower (5.8%) than in the peripheral retina (12.1%) (n = 9, P = 0.025, analysis of variance, P = 0.0074 for pairwise comparison). Confocal microscopy identified RGCs from the treated eyes with irregularly shaped cell soma, hypofluorescent cell soma, or that were lacking dendrites. Also observed were RGC dendrites that were abnormally thin or that appeared to be abruptly broken at primary branch points. Conclusions: Changes in RGC survival and structure following axon damage are readily assessed within the Thy1–CFP mouse. This transgenic mouse model may be useful for evaluating cellular mechanisms of RGC damage as well as potential new treatments to protect against RGC damage.

Keywords: ganglion cells • microscopy: confocal/tunneling • microscopy: light/fluorescence/immunohistochemistry 
© 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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