title: Förster resonance energy transfer as a tool to study photoreceptor biology.
abstract: Vision is initiated in photoreceptor cells of the retina by a set of biochemical events called phototransduction. These events occur via coordinated dynamic processes that include changes in secondary messenger concentrations, conformational changes and post-translational modifications of signaling proteins, and protein-protein interactions between signaling partners. A complete description of the orchestration of these dynamic processes is still unavailable. Described in this work is the first step in the development of tools combining fluorescent protein technology, Förster resonance energy transfer (FRET), and transgenic animals that have the potential to reveal important molecular insights about the dynamic processes occurring in photoreceptor cells. We characterize the fluorescent proteins SCFP3A and SYFP2 for use as a donor-acceptor pair in FRET assays, which will facilitate the visualization of dynamic processes in living cells. We also demonstrate the targeted expression of these fluorescent proteins to the rod photoreceptor cells of Xenopus laevis, and describe a general method for detecting FRET in these cells. The general approaches described here can address numerous types of questions related to phototransduction and photoreceptor biology by providing a platform to visualize dynamic processes in molecular detail within a native context.
authors.affiliation: Case Western Reserve University, Department of Ophthalmology and Visual Sciences, Cleveland, OH 44106, USA.
authors.forename: Stephanie C
authors.initials: SC
authors.lastname: Hovan
authors.forename: Scott
authors.initials: S
authors.lastname: Howell
authors.forename: Paul S-H
authors.initials: PS
authors.lastname: Park
mesh_terms: D000818:Animals; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D008027:Light; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D009865:Oocytes / Q000502:physiology* / Q000528:radiation effects*; D012243:Rhodopsin / Q000502:physiology*; D014982:Xenopus laevis
chemical_list: D012243:Rhodopsin