Live applications of norbormide-based fluorescent probes in Drosophila melanogaster

In this study we investigated the performance of two norbormide (NRB)-derived fluorescent probes, NRBMC009 (green) and NRBZLW0047 (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...

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Bibliographic Details
Published in:	PloS one Vol. 14; no. 4; p. e0211169
Main Authors:	Forgiarini, Alessia, Wang, Zifei, D'Amore, Claudio, Jay-Smith, Morgan, Li, Freda Fan, Hopkins, Brian, Brimble, Margaret Anne, Pagetta, Andrea, Bersani, Sara, De Martin, Sara, Napoli, Barbara, Bova, Sergio, Rennison, David, Orso, Genny
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 08-04-2019 Public Library of Science (PLoS)
Subjects:	Abnormalities Animals Biocompatibility Biology and Life Sciences Cell membranes Central nervous system Confocal microscopy Drosophila Drosophila melanogaster - anatomy & histology Drosophila melanogaster - physiology Dyes Eating Endoplasmic reticulum Epithelial cells Female Fluorescence Fluorescent Dyes - administration & dosage Fluorescent Dyes - chemistry Fluorescent Dyes - toxicity Fluorescent indicators Food intake Fruit flies Fruit flies (Tephritidae) Gastrointestinal Absorption Gastrointestinal Tract - diagnostic imaging Genes Green Fluorescent Proteins - chemistry Insects Internalization Internet Intravital Microscopy - methods Larva - physiology Larvae Life cycles Lipids Localization Male Markers Medicine and Health Sciences Microscopy Microscopy, Confocal Microscopy, Fluorescence Mitochondria Models, Animal Morphology Muscles Mutation Neurosciences Norbornanes - administration & dosage Norbornanes - chemistry Norbornanes - toxicity Pharmaceuticals Phenotypes Probes Proteins Pyridine Research and Analysis Methods Skeletal muscle Tissues Toxicity Toxicity Tests, Chronic New Zealand Italy Singapore
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Summary:	In this study we investigated the performance of two norbormide (NRB)-derived fluorescent probes, NRBMC009 (green) and NRBZLW0047 (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 NRBMC009 and NRBZLW0047 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 GFP-tagged protein markers, and a lack of toxicity make NRBZLW0047 and, particularly, NRBMC009 highly performing fluorescent probes for live cell microscopy studies and food intake assays in Drosophila.
Bibliography:	Competing Interests: The authors have declared that no competing interests exist.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0211169