A FRET-based cGMP biosensor in Drosophila
Resumen:
CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level.
| 2023 | |
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Agencia Nacional de Investigación e Innovación Dirección Nacional de Innovación, Ciencia y Tecnología |
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Drosophila FRET cGMP biosensor Ciencias Naturales y Exactas Ciencias Biológicas Biología Celular, Microbiología |
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| Inglés | |
| Instituto de Investigaciones Biológicas Clemente Estable | |
| IIBCE en REDI | |
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https://hdl.handle.net/20.500.12381/3367
https://www.doi.org/10.17912/micropub.biology.000887 |
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| Acceso abierto | |
| Reconocimiento 4.0 Internacional. (CC BY) |
| Sumario: | CUTie2 is a FRET-based cGMP biosensor tested so far only in cells. To expand its use to multicellular organisms we generated two transgenic Drosophila melanogaster strains that express the biosensor in a tissue-dependent manner. CUTie2 expression and subcellular localization was verified by confocal microscopy. The performance of CUTie2 was analyzed on dissected larval brains by hyperspectral microscopy and flow cytometry. Both approaches confirmed its responsivity, and the latter showed a rapid and reversible change in the fluorescence of the FRET acceptor upon cGMP treatment. This validated reporter system may prove valuable for studying cGMP signaling at organismal level. |
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