Purchase this article with an account.
David M. Gamm, Linda K. Barthel, Pamela A. Raymond, Michael D. Uhler; Localization of cGMP-Dependent Protein Kinase Isoforms in Mouse Eye. Invest. Ophthalmol. Vis. Sci. 2000;41(9):2766-2773.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
purpose. To examine the expression of the major isoforms of cyclic guanosine
monophosphate (cGMP)-dependent protein kinase (cGK) in mouse eye.
methods. Immunohistochemical localization of cGMP in mouse eye cryosections was
performed using an anti-cGMP antibody, followed by visualization with
indirect fluorescence microscopy. The presence of types Iα, Iβ, and
II cGK mRNAs in mouse eye extracts was determined initially by RNase
protection analysis. Further localization of cGK I and II mRNAs on
cryosections was accomplished by in situ hybridization using
digoxigenin-labeled cRNA probes and an alkaline phosphatase-conjugated
anti-digoxigenin antibody. Finally, cGK I protein was localized to
subcellular areas within the retina using an anti-cGK I–specific
results. In initial immunohistochemical experiments cGMP was present in
numerous regions and layers within the eye and retina. Subsequent RNase
protection studies demonstrated that cGK Iα, Iβ, and II mRNAs were
present in mouse eye and that type Iβ mRNA were 6.6 and 30 times more
abundant than type Iα and type II, respectively. By in situ
hybridization, cGK I mRNA was localized to photoreceptor inner segments
and the ganglion cell and inner nuclear layers of the retina, and
lesser amounts were found in the ciliary epithelium, lens, and cornea.
The cGK II mRNA expression pattern was similar but not identical with
that of cGK I. Finally, within the retina, cGK I protein was most
abundant in the inner plexiform layer, with significant amounts in
ganglion cells and photoreceptor inner segments as well.
conclusions. The presence of these cGK isoforms in discrete areas throughout the eye
suggests multiple roles for the cGMP-dependent signal transduction
system in the regulation of physiologic and pathologic ocular
This PDF is available to Subscribers Only