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Laurence Leconte, Colin J. Barnstable; Impairment of Rod cGMP-Gated Channel α-Subunit Expression Leads to Photoreceptor and Bipolar Cell Degeneration. Invest. Ophthalmol. Vis. Sci. 2000;41(3):917-926.
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purpose. To determine whether alterations in rod cGMP-gated channel function
mediate retinal degeneration, a transgenic approach in which the α
subunit of the rod cGMP-gated channel is reduced by the expression of
an antisense RNA was used.
methods. A 890-bp fragment of the 5′ mouse rod cGMP-gated channel cDNA was
cloned in the antisense orientation under the control of the strong
bacterial cytomegalovirus promoter. This antisense construct was used
to generate transgenic mice in which the expression of the antisense
transgene was measured by reverse transcription–polymerase chain
reaction. Histologic, immunocytochemical, and TdT-dUTP terminal
nick-end labeling (TUNEL) analyses were performed on control and
transgenic retina sections to determine the effects of antisense
expression on specific cell types.
results. The antisense RNA was able to inhibit the translation of the rod
channel protein in an in vitro assay. Three transgenic mouse lines
expressing the antisense construct were obtained. Molecular analyses
showed that the antisense is expressed in the eye of each transgenic
mouse line, where it reduces rod cGMP-gated channel RNA expression.
Histologic and immunocytochemical data showed that transgenic retinas
have a reduced number of photoreceptors and bipolar cells. TUNEL
staining confirmed that photoreceptor and bipolar cells degenerate
along an apoptotic pathway.
conclusions. Impairment of rod cGMP-gated channel α subunit expression leads to
photoreceptor and bipolar cell degeneration. These transgenic mice are
the first model of retinal degeneration caused by an alteration in the
expression of the rod cGMP-gated channel. This model system can be used
to test therapies designed to slow or stalled retinal
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