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E.C. Steele, Jr.; Analysis of L–Type Calcium Channel Cav1.3 Subunit Expression in the Mouse Retina . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1046.
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, To examine the distribution of the L–type calcium channel Cav1.3 subunit in the mouse retina.
For analysis of mRNA expression, in situ hybridization of mouse retinal sections was performed using Cav1.3–specific antisense and sense probes. For analysis of protein expression, a novel IgY antibody was generated in chicken against a unique and conserved Cav1.3 peptide sequence. This antibody was used in immunohistochemical analyses of mouse retinal sections and dissociated retinal cells and in Western blot analyses of mouse brain and retinal extracts.
In situ hybridization of mouse retinal sections revealed mRNA expression in all 3 nuclear layers. Immunohistochemical staining of mouse retinal sections and dissociated retinal cells with the anti–Cav1.3 antibody, but not non–immune IgY, revealed protein expression in specific retinal neuronal and glial cell types which was competed by preadsorption of the antibody with immunogen peptide. In Western blots of both mouse brain and mouse retinal cell extracts, the antibody recognized a band of appropriate molecular weight, which was competed by preadsorption of the antibody with immunogen peptide.
The expression profile of Cav1.3 in the mouse retina suggests an important role for this L–type calcium channel subunit in the regulation of both synaptic and extrasynaptic calcium–dependent processes in specific neurons and glial cells. In addition, the results confirm the utility and specificity of our anti–Cav1.3 anitbody which will greatly facilitate future investigations of the role of this channel subunit in the retina and in other tissues in which this channel is expressed.
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