Visualization of subcellular thermoregulation in live cells using fluorescent coiled-coil proteins

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In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration localized in different subcellular organelles have been proposed. However, previous studies have failed to observe thermogenesis directly in intact organelles. Here, we have visualized intracellular thermogenesis by using a fluorescent thermosensor, tsGFP. Specific targeting of tsGFPs enables visualization of thermogenesis in discrete organelles, notably mitochondria in brown adipocytes and the endoplasmic reticulum in myotubes. Strikingly, in HeLa cells, targeted tsGFP reveals heterogeneity in mitochondrial thermogenesis that is well correlated with the electrochemical gradient. Thus, tsGFPs directly demonstrate subcellular thermotransduction, and will serve as powerful tools to assess thermogenesis in vivo.