Retinal preconditioning and the induction of heat-shock protein 27

Brief periods of ischemia have been shown to protect the retina from potentially damaging periods of ischemia. This phenomenon has been termed ischemic preconditioning or ischemic tolerance. In the present study the cellular changes in levels of heat shock protein (Hsp)27, -70, and -90 mRNA and expression of Hsp in the rat retina associated with ischemic preconditioning were evaluated.

Unilateral retinal ischemia was created in Long-Evans and Sprague-Dawley rats for 5 minutes. Rats were then left for 1 hour to 7 days, to allow the retina to reperfuse. Retinas were dissected, the mRNA and protein isolated, and Northern and Western blot analyses conducted to detect changes in expression of Hsp27, -70, and -90. Immunohistochemical studies were used to identify retinal regions where Hsp changes occurred. Selected animals were subjected to a second ischemic event, 60 minutes in duration, to correlate the changes in expression of Hsp with functional protection of the retina from ischemic injury.

In control and sham-treated animals retinal Hsp27, -70, and -90 mRNAs were detectable. Five hours after retinal preconditioning, levels of Hsp27 mRNA were elevated above control levels, and 24 hours later, mRNA levels increased 200% over basal levels. Hsp27 expression remained elevated for up to 72 hours and then began to return to control levels. Hsp27 protein levels were increased by 200% over basal levels 24 hours after retinal preconditioning, remained at this level for 72 hours, and then returned to control levels. In contrast, no consistent change in Hsp70 or -90 mRNA or protein levels was observed during the course of the study. Immunohistochemical studies demonstrated that the increase in expression of Hsp27 was localized to neuronal and non-neuronal cells in the inner layers of the retina. Electroretinography studies demonstrated a strong correlation between the protection of retinal function from ischemic injury and the expression of Hsp27.

These results provide evidence that the induction of Hsp27 is a gene-specific event associated with ischemic preconditioning in the retina. This increase in expression of Hsp27 occurs in both neuronal and non-neuronal retinal cells, and appears to be one component of the neuroprotective events induced by ischemic preconditioning in the retina.