IOVS Journal of Pharmacology and Experimental Therapeutics
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(Investigative Ophthalmology and Visual Science. 2000;41:1159-1167.)
© 2000 by The Association for Research in Vision and Ophthalmology, Inc.

Characterization and Localization of the Rabbit Ocular Calcitonin Gene-Related Peptide (CGRP)-Receptor Component Protein (RCP)

Mark I. Rosenblatt1, Gerhard P. Dahl2 and Ian M. Dickerson1,2

From the Departments of 1 Biochemistry and Molecular Biology and 2 Physiology and Biophysics, the University of Miami School of Medicine, Miami, Florida.

PURPOSE. To determine whether the calcitonin gene-related peptide (CGRP) receptor component protein (RCP), a novel signal transduction molecule, is required for CGRP signaling in the eye and to determine potential ocular sites of CGRP action.

METHODS. The cDNA for the rabbit ocular RCP homologue was cloned using a combination of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Function of the rabbit ocular RCP was assessed using a sensitive oocyte-based assay, which utilizes the protein kinase A (PKA)-sensitive cystic fibrosis transmembrane conductance regulator (CFTR) as a sensor of cAMP formation. RCP expression in the rabbit eye was localized using immunohistochemistry.

RESULTS. A 2063-bp cDNA for the rabbit ocular RCP was cloned and sequenced. Expression of the rabbit RCP cDNA confers CGRP responsiveness in a sensitive oocyte-based assay. Antisense oligonucleotides made to the ocular RCP abolishes CGRP responsiveness of ciliary body and iris mRNA in the oocyte–CFTR assay. Localization of RCP protein in the rabbit eye using immunohistochemistry demonstrated RCP immunoreactivity in the ciliary body and iris blood vessels, as well as in layers of the ciliary epithelium.

CONCLUSIONS. The rabbit ocular RCP appears to be required for signal transduction at ocular CGRP receptors and is localized to sites previously reported to bind CGRP, which affect intraocular pressure and neurogenic inflammation.




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