Journal article
The challenges of using a copper fluorescent sensor (CS1) to track intracellular distributions of copper in neuronal and glial cells
KA Price, JL Hickey, Z Xiao, AG Wedd, SA James, JR Liddell, PJ Crouch, AR White, PS Donnelly
Chemical Science | ROYAL SOC CHEMISTRY | Published : 2012
DOI: 10.1039/c2sc20397a
Abstract
Copper is an essential biometal involved in critical cell functions including respiration. However, the mechanisms controlling its sub-cellular localization during health and disease remain poorly understood. This is partially due to the difficulty of detecting a metal ion that is bound tightly to metallo-chaperone and detoxification molecules in the cell. A BODIPY-based Cu fluorescent probe CS1 (Cu sensor 1) has been applied in innovative attempts to visualize monovalent Cu pools within cells (Zeng et al., J. Am. Chem. Soc., 2006, 128, 10-11). Inspired by this work, we sought to use CS1 to identify sub-cellular localization of Cu delivered to M17 neuronal or U87MG glial cells by a cell-perm..
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Funding Acknowledgements
This work was supported by funding from the National Health and Medical Research Council of Australia, the Australian Research Council and funds from the CSIRO OCE Postdoctoral Fellowship Program (SAJ). Aspects of this research were undertaken on the X-ray Fluorescence Microscopy beamline at the Australian Synchrotron. We would like to thank Prof. James Camakaris, Department of Genetics, The University of Melbourne for his valuable suggestions and input into this work. We thank Assoc. Prof. Andrew Hill and Dr Percy Chu, Department of Biochemistry and Molecular Biology, and Bio21 Institute, The University of Melbourne for help with live cell imaging. We also thank Dr Sharon La Fontaine, Dr Roxana Llanos and Mr Kenny Tran of Deakin University, Victoria, Australia for assistance with AAS, and Jens Brose and Lee Xin Chong of the University of Melbourne for isolation of Atox1 and CopK proteins.