Genetically encoded fluorescent thermosensors visualize subcellular thermoregulation in living cells
Genetically encoded sensors based on GFP enable the visualization of subcellular thermal changes noninvasively in intact cells. In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different sub...
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| Published in | Nature methods Vol. 10; no. 12; pp. 1232 - 1238 |
|---|---|
| Main Authors | , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
New York
Nature Publishing Group US
01.12.2013
Nature Publishing Group |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Genetically encoded sensors based on GFP enable the visualization of subcellular thermal changes noninvasively in intact cells.
In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the
Salmonella
thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells. |
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| AbstractList | In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the Salmonella thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells.In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the Salmonella thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells. Genetically encoded sensors based on GFP enable the visualization of subcellular thermal changes noninvasively in intact cells. In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the Salmonella thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells. In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the Salmonella thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells. |
| Audience | Academic |
| Author | Sakaguchi, Reiko Kiyonaka, Shigeki Omatsu-Kanbe, Mariko Imamura, Hiromi Morii, Takashi Hamachi, Itaru Kajimoto, Taketoshi Matsuura, Hiroshi Yoshizaki, Takenao Shinmi, Daisuke Mori, Yasuo |
| Author_xml | – sequence: 1 givenname: Shigeki surname: Kiyonaka fullname: Kiyonaka, Shigeki organization: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Department of Technology and Ecology, Hall of Global Environmental Studies, Kyoto University, Core Research for Evolution Science and Technology, Japan Science and Technology Agency – sequence: 2 givenname: Taketoshi surname: Kajimoto fullname: Kajimoto, Taketoshi organization: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University – sequence: 3 givenname: Reiko surname: Sakaguchi fullname: Sakaguchi, Reiko organization: World Premier International Research Initiative-Institute for Integrated Cell-Material Sciences, Kyoto University – sequence: 4 givenname: Daisuke surname: Shinmi fullname: Shinmi, Daisuke organization: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University – sequence: 5 givenname: Mariko surname: Omatsu-Kanbe fullname: Omatsu-Kanbe, Mariko organization: Department of Physiology, Shiga University of Medical Science – sequence: 6 givenname: Hiroshi surname: Matsuura fullname: Matsuura, Hiroshi organization: Department of Physiology, Shiga University of Medical Science – sequence: 7 givenname: Hiromi surname: Imamura fullname: Imamura, Hiromi organization: The Hakubi Project, Kyoto University – sequence: 8 givenname: Takenao surname: Yoshizaki fullname: Yoshizaki, Takenao organization: Department of Polymer Chemistry, Graduate School of Engineering, Kyoto University – sequence: 9 givenname: Itaru surname: Hamachi fullname: Hamachi, Itaru organization: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University – sequence: 10 givenname: Takashi surname: Morii fullname: Morii, Takashi organization: Institute of Advanced Energy, Kyoto University – sequence: 11 givenname: Yasuo surname: Mori fullname: Mori, Yasuo email: mori@sbchem.kyoto-u.ac.jp organization: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Department of Technology and Ecology, Hall of Global Environmental Studies, Kyoto University, Core Research for Evolution Science and Technology, Japan Science and Technology Agency |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24122038$$D View this record in MEDLINE/PubMed |
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| Snippet | Genetically encoded sensors based on GFP enable the visualization of subcellular thermal changes noninvasively in intact cells.
In mammals and birds,... In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in... |
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| SubjectTerms | 631/1647/1888/2249 631/443/319 631/80/2373 Adenoviridae - genetics Adipocytes, Brown - cytology Bacterial Proteins - chemistry Bioinformatics Biological Microscopy Biological Techniques Biomedical Engineering/Biotechnology Biosensors Birds Body temperature Cells DNA, Complementary - metabolism Electrochemistry Escherichia coli - metabolism Fluorescence Genetic aspects Genetics Green Fluorescent Proteins - chemistry HeLa Cells Heterogeneity Hot Temperature Humans Life Sciences Luminescent Proteins - metabolism Mammals Microscopy, Fluorescence - methods Mitochondria - metabolism Mitochondrial DNA Molecular biology Muscle Fibers, Skeletal - metabolism Muscle, Skeletal - metabolism Physiological aspects Protein Conformation Proteomics Regulation Salmonella Salmonella enterica - metabolism Sensors Temperature |
| Title | Genetically encoded fluorescent thermosensors visualize subcellular thermoregulation in living cells |
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