Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes

Rókus Kriszt, Satoshi Arai, Hideki Itoh, Michelle H. Lee, Anna G. Goralczyk, Xiu Min Ang, Aaron M. Cypess, Andrew P. White, Farnaz Shamsi, Ruidan Xue, Jung Yeol Lee, Sung Chan Lee, Yanyan Hou, Tetsuya Kitaguchi, Thankiah Sudhaharan, Shin'Ichi Ishiwata, E. Birgitte Lane, Young Tae Chang, Yu Hua Tseng, Madoka SuzukiMichael Raghunath

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Abstract

The identification of brown adipose deposits in adults has led to significant interest in targeting this metabolically active tissue for treatment of obesity and diabetes. Improved methods for the direct measurement of heat production as the signature function of brown adipocytes (BAs), particularly at the single cell level, would be of substantial benefit to these ongoing efforts. Here, we report the first application of a small molecule-Type thermosensitive fluorescent dye, ERthermAC, to monitor thermogenesis in BAs derived from murine brown fat precursors and in human brown fat cells differentiated from human neck brown preadipocytes. ERthermAC accumulated in the endoplasmic reticulum of BAs and displayed a marked change in fluorescence intensity in response to adrenergic stimulation of cells, which corresponded to temperature change. ERthermAC fluorescence intensity profiles were congruent with mitochondrial depolarisation events visualised by the JC-1 probe. Moreover, the averaged fluorescence intensity changes across a population of cells correlated well with dynamic changes such as thermal power, oxygen consumption, and extracellular acidification rates. These findings suggest ERthermAC as a promising new tool for studying thermogenic function in brown adipocytes of both murine and human origins.

Original languageEnglish
Article number1383
JournalScientific Reports
Volume7
Issue number1
DOIs
Publication statusPublished - 2017 Dec 1

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    Kriszt, R., Arai, S., Itoh, H., Lee, M. H., Goralczyk, A. G., Ang, X. M., Cypess, A. M., White, A. P., Shamsi, F., Xue, R., Lee, J. Y., Lee, S. C., Hou, Y., Kitaguchi, T., Sudhaharan, T., Ishiwata, SI., Lane, E. B., Chang, Y. T., Tseng, Y. H., ... Raghunath, M. (2017). Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes. Scientific Reports, 7(1), [1383]. https://doi.org/10.1038/s41598-017-00291-9