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Stress-related genomic responses during the course of heat acclimation and its association with ischemic-reperfusion cross-tolerance

¹Laboratory of Environmental Physiology, Faculty of Dental Medicine and ²School of Computer Science and Engineering, The Hebrew University, Jerusalem 91120, Israel

Submitted 22 March 2004 ; accepted in final form 14 May 2004

Acclimation to heat is a biphasic process involving a transient perturbed phase followed by a long lasting period during which acclimatory homeostasis is developed. In this investigation, we used cDNA stress microarray (Clontech Laboratory) to characterize the stress-related genomic response during the course of heat acclimation and to test the hypotheses that 1) heat acclimation influences the threshold of activation of protective molecular signaling, and 2) heat-acclimation-mediated ischemic-reperfusion (I/R) protection is coupled with reprogrammed gene expression leading to altered capacity or responsiveness of protective-signaling pathways shared by heat and I/R cytoprotective systems. Rats were acclimated at 34°C for 0, 2, and 30 days. ³²P-labeled RNA samples prepared from the left ventricles of rats before and after subjection to heat stress (HS; 2 h, 41°C) or after I/R insult (ischemia: 75%, 45 min; reperfusion: 30 min) were hybridized onto the array membranes. Confirmatory RT-PCR of selected genes conducted on samples taken at 0, 30, and 60 min after HS or total ischemia was used to assess the promptness of the transcriptional response. Cluster analysis of the expressed genes indicated that acclimation involves a "two-tier" defense strategy: an immediate transient response peaking at the initial acclimating phase to maintain DNA and cellular integrity, and a sustained response, correlated with slowly developed adaptive, long-lasting cytoprotective signaling networks involving genes encoding proteins that are essential for the heat-shock response, antiapoptosis, and antioxidation. Gene activation was stress specific. Faster activation and suppression of signaling pathways shared by HS and I/R stressors probably contribute to heat-acclimation I/R cross-tolerance.

ischemia-reperfusion ; stress genes; microarray; heart; heat stress

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