Heat shock response to hypoxia and its attenuation during recovery in the flesh fly, Sarcophaga crassipalpis

In this study, pharate adults of the flesh fly Sarcophaga crassipalpis were exposed to two, four, seven, or ten days of severe hypoxia (3% oxygen) to evaluate its impact on emergence and the expression of genes encoding heat shock proteins (Hsps) and heat shock regulatory elements. A four-day exposure to hypoxia significantly reduced survival, but more than seven days was required to reach the LD₅₀. Eight genes encoding Hsps, at least one from each major family of Hsps (Hsp90, Hsp70, Hsp60, Hsp40, and sHsps) and two genes encoding proteins involved in Hsp regulation (heat shock factor, hsf, and sirtuin) were cloned, and expression levels were assessed during and after hypoxia using qRT-PCR. Most, but not all hsps studied, were significantly up-regulated during hypoxia, and expression levels for most of the hsps reverted to control levels a few hours after return to normoxia. Hsp70 was the most responsive to hypoxia, increasing expression several hundred fold. By contrast, hsp90 and hsp27 showed little response to hypoxia but did respond to recovery. Neither hsf nor sirtuin were elevated by hypoxia, an observation consistent with their assumed post-transcriptional regulatory roles. These data demonstrate a strong Hsp response to hypoxia, suggesting an important role for Hsps in responding to low oxygen environments.