Abstract
The Drosophila visual system has been proved to be a powerful genetic model to study eye disease such as retinal degeneration. Here, we describe a genetic method termed “Rh1::GFP ey-flp/hid” that is based on the fluorescence of GFP-tagged major rhodopsin Rh1 in the eyes of living flies and can be used to monitor the integrity of photoreceptor cells. Through combination of this method and ERG recording, we examined a collection of 667 mutants and identified 18 genes that are required for photoreceptor cell maintenance, photoresponse, and rhodopsin synthesis. Our findings demonstrate that this “Rh1::GFP ey-flp/hid” method enables high-throughput F1 genetic screens to rapidly and precisely identify mutations of retinal degeneration.
Highlights
The Drosophila visual system has been shown to be proven to be a powerful genetic model for dissecting the molecular mechanisms underlying retinal degeneration and G-protein-coupled signaling cascades
Drosophila phototransduction offers the opportunity to combine classical and modern genetic approaches to identify genes and proteins that function in phototransduction and/or that are required for photoreceptor cell survival [1,2,3,4]
Electroretinogram recordings (ERGs) are among the analysis tools that have driven the progress of Drosophila phototransduction research; this technique is simple enough to be used to perform genetic screens [3]
Summary
The Drosophila visual system has been shown to be proven to be a powerful genetic model for dissecting the molecular mechanisms underlying retinal degeneration and G-protein-coupled signaling cascades. Phototaxis in the F1 generation is not sensitive enough, and the ERG-based high throughput screening is time-consuming [8,9,10] Given these nontrivial limitations, we were motivated to develop a fluorescence-based “Rh1::GFP ey-flp/hid” method for F1 screening of retinal degeneration mutants. We were motivated to develop a fluorescence-based “Rh1::GFP ey-flp/hid” method for F1 screening of retinal degeneration mutants This method is based on a modified EGUF/hid technique to generate eyes of homozygous mutations and uses GFP-tagged Rh1 (major rhodopsin) as a marker for photoreceptor cell integrity. Using this Rh1::GFP ey-flp/hid method, we screened the UCLA URCFG P-element recessive lethal collection, and identified several types of mutations affecting photoreceptor cell survival, phototransduction, and rhodopsin homeostasis
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