Heat shock protein 70 (Hsp70, Hspa1a) is known to play a protective role in the inner ear and in the nervous system. Our recent study demonstrated that the induction of Hsp70 by geldanamycin protected the auditory hair cells against ototoxic insult. Here, using the explanted organ of Corti (OC), we characterized the effect of sodium salicylate on the expression of Hsp70. Using the real-time RT-PCR; after 27 h in standard culture, we observed an increase in the Hsp70 transcript number. After 48 h in culture, the number of Hsp70 transcripts increased further, as compared to the freshly isolated tissues or explant cultured for 27 h. Three hours after the addition of 2.5mM sodium salicylate, the expression of Hsp70 mRNA increased significantly. Interestingly, Hsp70 protein level remained unaffected by the addition of salicylate, as shown by immunoblotting and Hsp70-ELISA. Confocal microscopy imaging demonstrated predominant localization of Hsp70 protein with or without salicylate exposure to the fibrocytes of spiral limbus. Our results suggest that in the OC, explanting process induces expression of Hsp70 in limbal fibrocytes and that this expression can be enhanced by salicylate but only on mRNA and not on the protein level.
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