Related Experiment Video
Updated: Dec 22, 2025
In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
Published on: August 20, 2019
Assaying Mutants of Clathrin-Mediated Endocytosis in the Fly Eye
Elsa Lauwers1, Patrik Verstreken2
1Laboratory of Neuronal Communication, KU Leuven, Department of Neurosciences, Leuven Brain Institute, VIB-KU Leuven Center for Brain & Disease Research, 49 Herestraat box 602, Leuven, 3000, Belgium. elsa.lauwers@kuleuven.vib.be.
Abstract:
Clathrin-mediated endocytosis plays essential roles both during and after development, and loss-of-function mutants affected in this process are mostly not viable. Different approaches have been developed to circumvent this limitation, including resorting to mosaic model organisms. We here describe the use of FLP/FRT-mediated mitotic recombination to generate Drosophila melanogaster having homozygous mutant eyes while the rest of their body is heterozygous. We then present a detailed protocol for assessing the consequences of these loss-of-function mutations on endocytosis in the photoreceptors of living fruit flies by recording electroretinograms.
Insights
We developed a method using FLP/FRT recombination in Drosophila to create eye-specific mutations in clathrin-mediated endocytosis. This allows studying essential gene functions in living flies by analyzing photoreceptor electroretinograms.
Area of Science:
- Cell Biology
- Genetics
- Developmental Biology
Background:
- Clathrin-mediated endocytosis is vital for cellular processes, but complete loss-of-function mutations are often lethal.
- Studying essential genes requires models that bypass organismal lethality.
Purpose of the Study:
- To develop a method for generating viable mosaic Drosophila melanogaster with homozygous mutant eyes for studying clathrin-mediated endocytosis.
- To assess the functional consequences of clathrin-mediated endocytosis loss-of-function in photoreceptors.
Main Methods:
- Utilized FLP/FRT-mediated mitotic recombination to generate mosaic flies.
- Created Drosophila eyes homozygous for endocytosis mutations while the rest of the body remained heterozygous.
- Recorded electroretinograms (ERGs) from living flies to assess photoreceptor function.
Main Results:
- Successfully generated mosaic Drosophila with homozygous mutant eyes for clathrin-mediated endocytosis genes.
- Demonstrated the feasibility of assessing endocytosis defects in photoreceptors using ERGs in living flies.
- Established a protocol for studying essential gene functions in a tissue-specific manner.
Conclusions:
- Mosaic analysis in Drosophila provides a powerful approach to study essential genes involved in clathrin-mediated endocytosis.
- Electroretinography is a viable method for assessing the functional impact of endocytosis mutations in vivo.
- This technique enables detailed investigation of developmental and cellular processes previously limited by lethality.
