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Peripheral thermosensation is the perception of external temperature. A change in temperature (on the surface of the skin and other tissues) is detected by a family of temperature-sensitive ion channels called Transient Receptor Potential, or TRP, receptors. These receptors are located on free nerve endings. Those detecting cold temperatures are closer to the surface of the skin than the nerve endings detecting warmth. These thermoTRP channels, while temperature selective, have relatively...
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A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like

Cong Quang Vu1,2, Shun-Ichi Fukushima2, Tetsuichi Wazawa2

  • 1Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.

Scientific Reports
|August 14, 2021
PubMed
Summary
This summary is machine-generated.

We developed ELP-TEMP, a highly sensitive genetically encoded temperature indicator (GETI), to measure real-time subcellular temperature dynamics in live cells. This new tool achieves unprecedented temperature sensitivity, enabling precise visualization of cellular heat production.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Molecular Imaging

Background:

  • Genetically encoded temperature indicators (GETIs) enable real-time measurement of subcellular temperature dynamics.
  • Existing GETIs often lack the sensitivity to detect small temperature changes from biological processes.
  • Accurate measurement of cellular temperature is crucial for understanding cell thermobiology.

Purpose of the Study:

  • To develop a highly sensitive genetically encoded temperature indicator (GETI) for precise subcellular temperature measurements.
  • To characterize the performance of the novel GETI, termed ELP-TEMP, in live cells.
  • To apply ELP-TEMP for visualizing heat production and measuring temperature changes in cellular compartments.

Main Methods:

  • Development of ELP-TEMP, a GETI utilizing a temperature-responsive elastin-like polypeptide (ELP) fused to Förster resonance energy transfer (FRET) pair (mTurquoise2 and mVenus).
  • Characterization of ELP-TEMP's temperature sensitivity and response range in HeLa cells.
  • Correction of ELP-TEMP output for macromolecular crowding and self-concentration effects.
  • Application of ELP-TEMP to detect localized temperature changes and visualize heat production from Ca2+ influx.

Main Results:

  • ELP-TEMP demonstrated a maximum temperature sensitivity of 45.1 ± 8.1%/°C between 33-40°C, significantly higher than previous fluorescent nanothermometers.
  • The indicator accurately measured temperature changes as small as <1°C induced by a local heat spot.
  • ELP-TEMP successfully visualized heat production from stimulated Ca2+ influx in live HeLa cells.
  • Temperatures in the nucleus and cytoplasm of HeLa cells were found to be nearly identical within the measurement resolution.

Conclusions:

  • ELP-TEMP represents a significant advancement in GETI technology, offering unparalleled temperature sensitivity for live-cell imaging.
  • The ability to accurately measure minute temperature changes and visualize heat production opens new avenues for studying cellular energetics.
  • ELP-TEMP is a valuable tool for investigating cellular temperature dynamics and advancing cell thermobiology research.