Live applications of norbormide-based fluorescent probes in Drosophila melanogaster

Alessia Forgiarini, Zifei Wang, Claudio D'Amore, Morgan Jay-Smith, Freda Fan Li, Brian Hopkins, Margaret Anne Brimble, Andrea Pagetta, Sara Bersani, Sara De Martin, Barbara Napoli, Sergio Bova, David Rennison, Genny Orso

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Abstract

In this study we investigated the performance of two norbormide (NRB)-derived fluorescent probes, NRB MC009 (green) and NRB ZLW0047 (red), on dissected, living larvae of Drosophila, to verify their potential application in live cell imaging confocal microscopy. To this end, larval tissues were exposed to NRB probes alone or in combination with other commercial dyes or GFP-tagged protein markers. Both probes were rapidly internalized by most tissues (except the central nervous system) allowing each organ in the microscope field to be readily distinguished at low magnification. At the cellular level, the probes showed a very similar distribution (except for fat bodies), defined by loss of signal in the nucleus and plasma membrane, and a preferential localization to endoplasmic reticulum (ER) and mitochondria. They also recognized ER and mitochondrial phenotypes in the skeletal muscles of fruit fly models that had loss of function mutations in the atlastin and mitofusin genes, suggesting NRB MC009 and NRB ZLW0047 as potentially useful screening tools for characterizing ER and mitochondria morphological alterations. Feeding of larvae and adult Drosophilae with the NRB-derived dyes led to staining of the gut and its epithelial cells, revealing a potential role in food intake assays. In addition, when flies were exposed to either dye over their entire life cycle no apparent functional or morphological abnormalities were detected. Rapid internalization, a bright signal, a compatibility with other available fluorescent probes and GFPtagged protein markers, and a lack of toxicity make NRB ZLW0047 and, particularly, NRB MC009 highly performing fluorescent probes for live cell microscopy studies and food intake assays in Drosophila.

Original languageEnglish
Article numbere0211169
Number of pages18
JournalPLoS One
Volume14
Issue number4
DOIs
Publication statusPublished - Apr 1 2019

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Forgiarini, A., Wang, Z., D'Amore, C., Jay-Smith, M., Li, F. F., Hopkins, B., Brimble, M. A., Pagetta, A., Bersani, S., De Martin, S., Napoli, B., Bova, S., Rennison, D., & Orso, G. (2019). Live applications of norbormide-based fluorescent probes in Drosophila melanogaster. PLoS One, 14(4), [e0211169]. https://doi.org/10.1371/journal.pone.0211169