June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Expression of Olfactory Receptor Genes on Mouse Ocular Surface
Author Affiliations & Notes
  • Vladlen Slepak
    Molecular and Cellular Pharmacology, University of Miami, Miami, FL
  • Alexey Pronin
    Molecular and Cellular Pharmacology, University of Miami, Miami, FL
  • Konstantin Levay
    Molecular and Cellular Pharmacology, University of Miami, Miami, FL
  • Yaohong Tan
    Opthalmology, University of Miami, Miami, FL
  • Dmitry Velmeshev
    Psychiatry, University of Miami, Miami, FL
  • Mohammad Faghihi
    Psychiatry, University of Miami, Miami, FL
  • Valery Shestopalov
    Opthalmology, University of Miami, Miami, FL
  • Footnotes
    Commercial Relationships Vladlen Slepak, None; Alexey Pronin, None; Konstantin Levay, None; Yaohong Tan, None; Dmitry Velmeshev, None; Mohammad Faghihi, None; Valery Shestopalov, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2579. doi:
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    • Get Citation

      Vladlen Slepak, Alexey Pronin, Konstantin Levay, Yaohong Tan, Dmitry Velmeshev, Mohammad Faghihi, Valery Shestopalov; Expression of Olfactory Receptor Genes on Mouse Ocular Surface. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2579.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

Ocular surface is continuously exposed to tear components as well as xenobiotics and commensal and pathogenic microorganisms. To advance our understanding how the outer eye interacts with its environment, we asked which cellular receptors are expressed on the ocular surface, focusing on identification of genes encoding G protein-coupled receptors (GPCRs).

 
Methods
 

Total RNA was isolated from mouse cornea and conjunctiva and subjected to next-generation sequencing using Illumina platform. The data was analyzed by CuffLinks and TopHat software packages. Gene expression was confirmed by RT-PCR using total or poly-adenylated RNA obtained from independent cornea and conjunctiva preparations. Cellular localization of transcripts was analyzed by in situ hybridization using RNAscope technology on paraffin-embedded slices of mouse eyes.

 
Results
 

We performed directional RNA deep sequencing, utilizing Epicenter library preparation kit to generate more than 45 million reads representing over 10,000 transcripts. We identified more than 150 GPCR gene transcripts, of which more than 100 were putative olfactory receptors (Olfr’s). Our RT-PCR analyses confirmed the presence of several GPCR and G protein transcripts, including the G protein associated with olfaction, Golf. In contrast, a number of control genes, such as insulin or rhodopsin did no show significant expression. In situ hybridization showed that mRNA for Olfr558 was present primarily in basal epithelial cells of cornea and conjunctiva.

 
Conclusions
 

Several species of mRNA encoding putative olfactory receptors are expressed on the mouse OS.

  
Keywords: 675 receptors: pharmacology/physiology • 480 cornea: basic science • 533 gene/expression  
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