Triclosan is a mitochondrial uncoupler in live zebrafish.
Triclosan (TCS) is a synthetic antimicrobial agent used in many consumer goods at millimolar concentrations. As a result of exposure, TCS has been detected widely in humans. We have recently discovered that TCS is a proton ionophore mitochondrial uncoupler in multiple types of living cells. Here, we present novel data indicating that TCS is also a mitochondrial uncoupler in a living organism: 24-hour post-fertilization (hpf) zebrafish embryos. These experiments were conducted using a Seahorse Bioscience XFe96 Extracellular Flux Analyzer modified for bidirectional temperature control, using the XF96 spheroid plate to position and measure one zebrafish embryo per well. Using this method, after acute exposure to TCS, the basal oxygen consumption rate (OCR) increases, without a decrease in survival or heartbeat rate. TCS also decreases ATP-linked respiration and spare respiratory capacity and increases proton leak: all indicators of mitochondrial uncoupling. Our data indicate, that TCS is a mitochondrial uncoupler in vivo, which should be taken into consideration when assessing the toxicity and/or pharmaceutical uses of TCS. This is the first example of usage of a Seahorse Extracellular Flux Analyzer to measure bioenergetic flux of a single zebrafish embryo per well in a 96-well assay format. The method developed in this study provides a high-throughput tool to identify previously unknown mitochondrial uncouplers in a living organism. Copyright © 2016 John Wiley & Sons, Ltd.
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.
Juyoung
J
Shim
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.|Graduate School of Biomedical Science and Engineering, University of Maine, Orono, Maine, 04469, USA.
Lisa M
LM
Weatherly
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.
Richard H
RH
Luc
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.
Maxwell T
MT
Dorman
Seahorse Bioscience, Inc., North Billerica, Massachusetts, 01862, USA.
Andy
A
Neilson
Seahorse Bioscience, Inc., North Billerica, Massachusetts, 01862, USA.
Ryan
R
Ng
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.|Graduate School of Biomedical Science and Engineering, University of Maine, Orono, Maine, 04469, USA.
Carol H
CH
Kim
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA.|Graduate School of Biomedical Science and Engineering, University of Maine, Orono, Maine, 04469, USA.|Department of Chemical and Biological Engineering and the Laboratory for Surface Science & Technology, University of Maine, Orono, Maine, 04469, USA.
Paul J
PJ
Millard
Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, 04469, USA. Julie.gosse@maine.edu.|Graduate School of Biomedical Science and Engineering, University of Maine, Orono, Maine, 04469, USA. Julie.gosse@maine.edu.
Julie A
JA
Gosse
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000187:drug effects*; D004785:Environmental Pollutants / Q000633:toxicity*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D011522:Protons; D014260:Triclosan / Q000633:toxicity*; D014475:Uncoupling Agents / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology / Q000378:metabolism
D004785:Environmental Pollutants; D011522:Protons; D014475:Uncoupling Agents; D014260:Triclosan
ATP production; Extracellular Flux Analyzer; bioenergetics; irgasan; mitochondrial uncoupler; oxygen consumption rate; proton leak; seahorse bioscience; triclosan; zebrafish