Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes.
Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes.
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DOI：
10.1038/s41598-017-00291-9
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发表时间：
2017-05-03
影响因子：
4.6
通讯作者：
Raghunath M
中科院分区：
文献类型：
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作者：
Kriszt R;Arai S;Itoh H;Lee MH;Goralczyk AG;Ang XM;Cypess AM;White AP;Shamsi F;Xue R;Lee JY;Lee SC;Hou Y;Kitaguchi T;Sudhaharan T;Ishiwata S;Lane EB;Chang YT;Tseng YH;Suzuki M;Raghunath M
成人中棕色脂肪沉积的鉴定已经引起了对靶向这种代谢活性组织用于治疗肥胖症和糖尿病的极大兴趣。用于直接测量作为棕色脂肪细胞（BA）的特征功能的产热的改进方法，特别是在单细胞水平，将对这些正在进行的努力具有实质性益处。在这里，我们报告的第一个应用程序的小分子型热敏荧光染料，ERthermAC，监测产热在BA来自小鼠棕色脂肪前体和人类棕色脂肪细胞分化的人颈棕色前脂肪细胞。ERthermAC在BAs的内质网中积累，并显示响应于肾上腺素能刺激细胞的荧光强度的显著变化，这对应于温度的变化。ERthermAC荧光强度曲线与JC-1探针观察到的线粒体去极化事件一致。此外，整个细胞群体的平均荧光强度变化与动态变化（如热功率、耗氧量和细胞外酸化速率）密切相关。这些研究结果表明ERthermAC作为一个有前途的新工具，研究生热功能的棕色脂肪细胞的小鼠和人类的起源。
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.