Purchase this article with an account.
Chooi-May Lai, Wei-Yong Shen, Ian Constable, Piroska Elizabeth Rakoczy; The Use of Adenovirus-Mediated Gene Transfer to Develop a Rat Model for Photoreceptor Degeneration. Invest. Ophthalmol. Vis. Sci. 2000;41(2):580-584.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
purpose. To investigate the effects of recombinant adenovirus-mediated
downregulation of cathepsin S (CatS) on the retinal pigment epithelium
and/or neural retina in vivo.
methods. The expression of green fluorescent protein (gfp) after subretinal
injection of a recombinant adenovirus, Ad.gfp, into rat eyes was first
established by in vivo fundus fluorescence photography and fluorescence
microscopy. The autofluorescent debris accumulation in Ad.CatSAS
(recombinant adenovirus carrying the antisense CatS gene)-injected rat
eyes was monitored by fluorescence microscopy, and the antisense CatS
RNA expression was demonstrated by in situ hybridization. Changes in
the retinal morphology were assessed by light microscopy.
results. The gfp expression was present in 30% to 90% of the injection area at
3 days and was absent 9 days after Ad.gfp injection. In
Ad.CatSAS-injected eyes, the expression of antisense CatS RNA was
demonstrated by in situ hybridization. Autofluorescent debris
accumulation was significantly higher in Ad.CatSAS-injected eyes than
in control eyes. The shortening of photoreceptor outer segments in
Ad.CatSAS-injected eyes coincided with intense autofluorescent debris
accumulation. The number of layers of photoreceptor cells decreased
with time and were 11, 9, and 8 at 7, 14, and 28 days after Ad.CatSAS
injection, respectively. In control eyes, the number of layers of
photoreceptor cells (14) remained unchanged.
conclusions. These results demonstrate that recombinant adenovirus-mediated
transient modulation of gene expression in retinal pigment epithelial
(RPE) cells could induce changes in the retina, and, in spite of the
low expression of endogenous CatS in RPE cells, this enzyme plays an
important role in maintenance of normal retinal
This PDF is available to Subscribers Only