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A FRET-based cGMP biosensor in Drosophila.
Ana Clara Gonzalez1, Cecilia Abreu2, Sergio Pantano3
1Departamento de Biología del Neurodesarrollo, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Montevideo, Uruguay.
Micropublication Biology
|December 14, 2023
Summary
Researchers developed transgenic Drosophila strains expressing the CUTie2 biosensor for studying cyclic guanosine monophosphate (cGMP) signaling in whole organisms. This FRET-based tool shows rapid, reversible responses, validating its use in multicellular contexts.
Area of Science:
- Molecular Biology
- Neuroscience
- Genetics
Background:
- Cyclic guanosine monophosphate (cGMP) is a crucial second messenger in various cellular processes.
- Existing biosensors like CUTie2 are primarily validated in cell cultures, limiting their application in complex organisms.
- Studying cGMP dynamics in vivo is essential for understanding its role in physiological and pathological conditions.
Purpose of the Study:
- To generate and validate transgenic *Drosophila melanogaster* models expressing the FRET-based cGMP biosensor CUTie2.
- To assess the tissue-specific expression, localization, and performance of CUTie2 in a multicellular organism.
- To establish a reliable reporter system for studying cGMP signaling in vivo.
Main Methods:
- Generation of two transgenic *Drosophila melanogaster* strains for tissue-dependent CUTie2 expression.
- Confocal microscopy for verifying biosensor expression and subcellular localization.
- Hyperspectral microscopy and flow cytometry on dissected larval brains to analyze CUTie2 performance and cGMP-induced responses.
Main Results:
- Successful generation of transgenic *Drosophila* strains expressing CUTie2 in a tissue-specific manner.
- Confocal microscopy confirmed proper expression and localization of the biosensor.
- Hyperspectral microscopy and flow cytometry demonstrated CUTie2's responsivity to cGMP, with flow cytometry showing rapid, reversible fluorescence changes.
Conclusions:
- The transgenic *Drosophila* models successfully express the CUTie2 cGMP biosensor.
- CUTie2 is validated as a functional reporter for cGMP signaling in a multicellular organism (*Drosophila*).
- This validated reporter system holds potential for advancing the study of cGMP signaling in vivo.
