Photoreceptor responses of fruitflies with normal and reduced arrestin content studied by simultaneous measurements of visual pigment fluorescence and ERG.
We have simultaneously measured the electroretinogram (ERG) and the metarhodopsin content via fluorescence in white-eyed, wild-type Drosophila and the arrestin2 hypomorphic mutant (w(-);arr2 (3)) at a range of stimulus wavelengths and intensities. Photoreceptor response amplitude and termination (transition between full repolarization and prolonged depolarizing afterpotential, PDA) were related to visual pigment conversions and arrestin concentration. The data were implemented in a kinetic model of the rhodopsin-arrestin cycle, allowing us to estimate the active metarhodopsin concentration as a function of effective light intensity and arrestin concentration. Arrestin reduction in the mutant modestly increased the light sensitivity and decreased the photoreceptor dynamic range. Compared to the wild type, in the mutant the transition between full repolarization and PDA occurred at a lower metarhodopsin fraction and was more abrupt. We developed a steady-state stochastic model to interpret the dependence of the PDA on effective light intensity and arrestin content and to help deduce the arrestin to rhodopsin ratio from the sensitivity and PDA data. The feasibility of different experimental methods for the estimation of arrestin content from ERG and PDA is discussed.
Belusic
Gregor
G
Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia. gregor.belusic@bf.uni-lj.si
Pirih
Primoz
P
Stavenga
Doekele G
DG
D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D003198:Computer Simulation; D015536:Down-Regulation / Q000235:genetics / Q000528:radiation effects; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D004331:Drosophila melanogaster / Q000166:cytology / Q000502:physiology*; D004596:Electroretinography; D005123:Eye; D005453:Fluorescence; D008564:Membrane Potentials / Q000235:genetics; D009154:Mutation / Q000235:genetics; D009799:Ocular Physiological Phenomena; D010775:Photic Stimulation; D017956:Photoreceptor Cells, Invertebrate / Q000166:cytology / Q000502:physiology* / Q000528:radiation effects; D012168:Retinal Pigments / Q000032:analysis / Q000378:metabolism / Q000528:radiation effects; D012243:Rhodopsin / Q000032:analysis / Q000378:metabolism; D013269:Stochastic Processes; D014785:Vision, Ocular / Q000235:genetics* / Q000528:radiation effects
C504666:Arr2 protein, Drosophila; D019390:Arrestins; D029721:Drosophila Proteins; D012168:Retinal Pigments; C013110:metarhodopsins; D012243:Rhodopsin