Heterologous expression of bovine rhodopsin in Drosophila photoreceptor cells

Syed Tariq Ahmad; Michael NATOCHIN; Brandy BARREN; Nikolai O Artemyev; Joseph E O'TOUSA

doi:10.1167/iovs.06-0281

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Heterologous expression of bovine rhodopsin in Drosophila photoreceptor cells

Journal article

Peer reviewed

Heterologous expression of bovine rhodopsin in Drosophila photoreceptor cells

Syed Tariq Ahmad, Michael NATOCHIN, Brandy BARREN, Nikolai O Artemyev and Joseph E O'TOUSA

Investigative ophthalmology & visual science, Vol.47(9), pp.3722-3728

2006

DOI: 10.1167/iovs.06-0281

PMID: 16936079

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Abstract

purpose. Vertebrate and invertebrate visual pigments are similar in amino acid sequence, structural organization, spectral properties, and mechanism of action, but possess different chromophores and trigger phototransduction through distinct biochemical pathways. The bovine opsin gene (Rho) was expressed in Drosophila, to examine the properties of a vertebrate opsin within invertebrate photoreceptor cells. methods. Transgenic Drosophila expressing the bovine opsin gene (Rho) in photoreceptors were created. Protein expression and cellular location of bovine rhodopsin was assessed by protein blots and immunofluorescence. The glycosylation state was determined by mobility profiles in SDS-PAGE before and after treatment with endoglycosidase. The rhodopsin chromophore was determined by HPLC-mass spectroscopy (MS) and the spectral properties by spectroscopy. The ability of the bovine rhodopsin to couple to Drosophila phototransduction components was assessed by electroretinography and to couple to vertebrate transducin by light-mediated GTPγS-binding assays. results. Rho showed stable expression even in the absence of endogenous Rh1 opsin and chromophore. It was correctly targeted to the rhabdomeric membranes. Rho remained glycosylated during the maturation process and possessed a distinct glycosylation pattern from that of native Rho. The Drosophila-expressed Rho associated with the 3-hydroxyretinal chromophore but failed to evoke an electroretinogram response from fly photoreceptors. However, the Drosophila-expressed Rho activated transducin in a light-dependent manner. conclusions. Drosophila photoreceptors express a vertebrate rhodopsin as a functional visual pigment, but the expression does not activate the Drosophila phototransduction pathway. The system allows the characterization and comparison of vertebrate and invertebrate visual pigment properties in a common cell type.

Fundamental and applied biological sciences. Psychology

Vertebrates: nervous system and sense organs

Biological and medical sciences

Eye and associated structures. Visual pathways and centers. Vision

Details

Title: Subtitle: Heterologous expression of bovine rhodopsin in Drosophila photoreceptor cells
Creators: Syed Tariq Ahmad - Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, United States
Michael NATOCHIN - Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa, United States
Brandy BARREN - Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa, United States
Nikolai O Artemyev - Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa, United States
Joseph E O'TOUSA - Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, United States
Resource Type: Journal article
Publication Details: Investigative ophthalmology & visual science, Vol.47(9), pp.3722-3728
DOI: 10.1167/iovs.06-0281
PMID: 16936079
NLM abbreviation: Invest Ophthalmol Vis Sci
ISSN: 0146-0404
eISSN: 1552-5783
Publisher: Association for Research in Vision and Ophtalmology; Rockville, MD
Language: English
Date published: 2006
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
Record Identifier: 9984070614802771

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