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Expression of olfactory GPCR signaling genes in the eye (662.15)
Author(s) -
Slepak Vladlen,
Pronin Alexey,
Levay Konstantin,
Velmeshev Dmitry,
Faghihi Mohammad,
Shestopalov Valery
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.662.15
Subject(s) - biology , g protein coupled receptor , cornea , in situ hybridization , gene , transcriptome , gene expression , receptor , messenger rna , microbiology and biotechnology , genetics , neuroscience
Ocular surface is continuously exposed to particles, microorganisms and other stressors. To advance our understanding how the outer eye interacts with its environment, we asked which cellular receptors are expressed in the cornea, focusing on identification of G protein‐coupled receptors (GPCRs). Total RNA was isolated from the mouse cornea and subjected to next‐generation sequencing using Illumina platform. The data was analyzed by CuffLinks and TopHat software. Gene expression was confirmed by RT‐PCR. Cellular localization of transcripts was analyzed by in situ hybridization using RNAscope technology. The determined mouse cornea transcriptome contained more than 45 million reads representing over 10,000 gene transcripts. We identified more than 150 GPCR gene transcripts including more than 100 were putative olfactory receptors (Olfr’s). Our RT‐PCR analysis confirmed the presence of several Olfrs and related genes, for example, the G protein associated with olfaction, Galpha‐olf, and olfactory marker protein (OMP). In situ hybridization showed that mRNA for Olfr558 was present primarily in vessels of the eye, whereas OMP was found in epithelial cells of the cornea. Thus, our data show that several species of mRNA encoding putative olfactory receptors and related genes are expressed in the mouse cornea and other parts of the eye. Grant Funding Source : Supported by NIH RO1EY018666 (VZS), R01EY017991 (VIS), P30 EY014801