Article
Version 3
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Versatile Cell and Animal Models for Advanced Investigation of Lead Poisoning
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and
Version 1
: Received: 23 August 2021 / Approved: 25 August 2021 / Online: 25 August 2021 (10:39:42 CEST)
Version 2 : Received: 17 September 2021 / Approved: 17 September 2021 / Online: 17 September 2021 (11:58:04 CEST)
Version 3 : Received: 1 October 2021 / Approved: 1 October 2021 / Online: 1 October 2021 (11:12:13 CEST)
Version 2 : Received: 17 September 2021 / Approved: 17 September 2021 / Online: 17 September 2021 (11:58:04 CEST)
Version 3 : Received: 1 October 2021 / Approved: 1 October 2021 / Online: 1 October 2021 (11:12:13 CEST)
A peer-reviewed article of this Preprint also exists.
Yang, D.-M.; Chang, Y.-F. Versatile Cell and Animal Models for Advanced Investigation of Lead Poisoning. Biosensors 2021, 11, 371. Yang, D.-M.; Chang, Y.-F. Versatile Cell and Animal Models for Advanced Investigation of Lead Poisoning. Biosensors 2021, 11, 371.
Abstract
The heavy metal lead (Pb) can irreversibly damage the human nervous system. To help understand Pb-induced damage, we applied a genetically encoded Förster resonance energy transfer (FRET)-based Pb biosensor Met-lead 1.44 M1 to two living systems to monitor the concentration of Pb: induced pluripotent stem cell (iPSC)-derived cardiomyocytes as a semi-tissue platform, and Drosophila melanogaster fruit flies as an in vivo animal model. Different FRET imaging modalities were used to obtain FRET signals, which represented the presence of Pb in the tested samples in different spatial dimensions. Using iPSC-derived cardiomyocytes, the relationship between beating activity (20–24 beats per minute, bpm) determined from the fluctuation of fluorescent signals and the concentrations of Pb represented by the FRET emission ratio values of Met-lead 1.44 M1 was revealed from simultaneous measurements. Pb (50 μM) affected the beating activity of cardiomyocytes, whereas two drugs that stop the entry of Pb differentially affected this beating activity: verapamil (2 μM) did not reverse the cessation of beating, whereas 2-APB (50 μM) partially restored this activity (16 bpm). The results clearly demonstrate a potential of this biosensor system as an anti-Pb drug screening application. In the Drosophila model, Pb was detected within the adult brain or larval central nervous system (Cha-gal4>UAS-Met-lead 1.44 M1) using fast epifluorescence and high-resolution two-photon 3D FRET ratio image systems. The tissue-specific expression of Pb biosensors provides an excellent opportunity to explore the possible Pb-specific populations within living organisms. We believe that this integrated Pb biosensor system can be applied to the prevention of Pb poisoning and advanced research on Pb neurotoxicology.
Keywords
Drosophila melanogaster; fluorescence resonance energy transfer; Met-lead 1.44 M1; Pb biosensor
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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