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A genetic reporter of thermal stress defines physiologic zones over a defined temperature range
Author(s) -
O'connellRodwell Caitlin E.,
Shriver David,
Simanovskii Dmitri M.,
Mcclure Cameron,
Cao Yuan,
Zhang Weisheng,
Bachmann Michael H.,
Beckham Joshua T.,
Jansen E. Duco,
Palanker Daniel,
Schwettman H. Alan,
Contag Christopher H.
Publication year - 2004
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.03-0585com
Subject(s) - reporter gene , luciferase , hsp70 , heat shock protein , gene expression , heat shock , microbiology and biotechnology , biology , gene , chemistry , biophysics , transfection , biochemistry
We define five unique cellular responses to thermal stress using a reporter construct generated using the stress‐inducible promoter from the gene encoding a murine 70 kDa heat shock protein (Hsp70A.1) to express luciferase ( luc ). Thermal stress was delivered over a range of temperatures (42–68°C) for5sto20min and luciferase activity was measured in live cells using a cooled CCD camera as a measure of reporter gene transcription. Reporter gene expression was assessed every 2 h for 10 h, and at 24 h post‐stress. Expression patterns were validated for selected tem¬peratures. A transition zone where cells lose the ability to produce light and beyond which >50% of cells die was observed to occur within a narrow (2.5°C) temper¬ature window. Although luc and hsp70 mRNA levels in this transition zone were high, there were reduced levels of Luc and Hsp70 protein and ATP levels. Cells treated at these temperatures recovered the ability to produce light in response to a secondary stress at 30 h. This Hsp70‐luc reporter gene construct may be useful for defining zones of physiologic responses and assess¬ing collateral thermal damage generated during treat¬ment of biological tissue with lasers and other sources of heat.—O'Connell‐Rodwell, C. E., Shriver, D., Simanovskii, D. M., McClure, C., Cao, Y‐a., Zhang, W., Bachmann, M. H., Beckham, J. T., Jansen, E. D., Palanker, D., Schwettman, H. A., Contag, C. H. A genetic reporter of thermal stress defines physiologic zones over a defined temperature range. FASEB J. 18, 264–271 (2004)