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Sci Rep. 2017 May 3;7(1):1383. doi: 10.1038/s41598-017-00291-9.

Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes.

Kriszt R1,2,3, Arai S4,5, Itoh H6,7, Lee MH2, Goralczyk AG1,2, Ang XM1,2,3, Cypess AM8,9, White AP10, Shamsi F8, Xue R8, Lee JY11, Lee SC12,13, Hou Y4, Kitaguchi T4,5, Sudhaharan T14, Ishiwata S4,5,15, Lane EB7, Chang YT11,12, Tseng YH16, Suzuki M17,18,19, Raghunath M20,21,22,23.

Author information

1
Department of Biomedical Engineering, National University of Singapore, 117583, Singapore, Singapore.
2
NUS Tissue Engineering Program, Life Science Institute, National University of Singapore, 117510, Singapore, Singapore.
3
NUS Graduate School for Integrative Sciences and Engineering (NGS), National University of Singapore, 117456, Singapore, Singapore.
4
WASEDA Bioscience Research Institute in Singapore (WABIOS), 138667, Singapore, Singapore.
5
Organization for University Research Initiatives, Waseda University, Tokyo, 162-0041, Japan.
6
Department of Pure and Applied Physics, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, 169-8555, Japan.
7
Epithelial Biology Laboratory, Institute of Medical Biology (IMB), Agency for Science, Technology and Research (A*STAR), 138648, Singapore, Singapore.
8
Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, MA, 02215, USA.
9
Diabetes, Endocrinology, and Obesity Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MA, 20892, USA.
10
Department of Orthopedic Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, 02215, USA.
11
Department of Chemistry & MedChem Program of Life Sciences Institute, National University of Singapore, 117543, Singapore, Singapore.
12
Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium (SBIC), Agency for Science, Technology and Research (A*STAR), 138667, Singapore, Singapore.
13
Aptabio Therapeutics Inc., Yongin City, 446-908, Korea.
14
Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), 138648, Singapore, Singapore.
15
Department of Physics, Faculty of Science and Engineering, Waseda University, Tokyo, 169-8555, Japan.
16
Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, MA, 02215, USA. yu-hua.tseng@joslin.harvard.edu.
17
WASEDA Bioscience Research Institute in Singapore (WABIOS), 138667, Singapore, Singapore. suzu_mado@aoni.waseda.jp.
18
Organization for University Research Initiatives, Waseda University, Tokyo, 162-0041, Japan. suzu_mado@aoni.waseda.jp.
19
PRESTO, Japan Science and Technology Agency, Saitama, 332-0012, Japan. suzu_mado@aoni.waseda.jp.
20
Department of Biomedical Engineering, National University of Singapore, 117583, Singapore, Singapore. ragh@zhaw.ch.
21
NUS Tissue Engineering Program, Life Science Institute, National University of Singapore, 117510, Singapore, Singapore. ragh@zhaw.ch.
22
Department of Biochemistry, Yong Loo Ling School of Medicine, National University of Singapore, 117597, Singapore, Singapore. ragh@zhaw.ch.
23
Institute for Chemistry and Biotechnology (ICBT), Zurich University of Applied Sciences, Wädenswil, CH - 8820, Switzerland. ragh@zhaw.ch.

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.

PMID:
28469146
PMCID:
PMC5431191
DOI:
10.1038/s41598-017-00291-9
[Indexed for MEDLINE]
Free PMC Article

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