g, phosphorylated GSK3B. Specifically, AKT phosphorylates GSK3B at ser9 and selleck Vandetanib the P GSK3B, leading to decreased activ ity of GSK3B. Activation of the AKTGSK3B signaling pathway, demon strated as increases in the levels of P AKT and P GSK3B, has been reported to protect against myocar dial Inhibitors,Modulators,Libraries apoptosis induced by ischemiareperfusion in rats and to increase the survival of hippocampal neurons fol lowing ischemia in rats. We therefore set out to investigate the potential dual effects of sevoflurane anesthesia on the AKTGSK3B sig naling pathway in young wild type mice and in H4 human neuroglioma cells. The hy pothesis in the study is that single exposure or short duration treatment with sevoflurane increases, but mul tiple exposures or long duration treatment with sevoflur ane decreases, the levels of P GSK3B and P AKT.

We used 3% sevoflurane in the animal studies because our previous studies showed that anesthesia with 3% sevo flurane two hours daily for three days, but not one day, was able to induce neuroinflammation and cognitive im pairment in mice. We used 4% sevoflurane in the in vitro studies because our previous in vitro studies showed the treatment with 4% sevoflurane for Inhibitors,Modulators,Libraries six hours could induce apoptosis and increase AB levels in the hu man H4 neuroglioma cells. Materials and Inhibitors,Modulators,Libraries methods Mice anesthesia and treatment All experiments were performed in accordance with the National Institute of Health guidelines and regulations. The animal protocol was approved by the Massachusetts Gen eral Hospital Standing Committee on the Use of Animals in Research and Teaching.

Efforts were made to minimize the number of animals used. Both male and female mice were used in the studies. Young mice were used in the current studies. The mice were randomly assigned into the anesthesia group or the control group. The mice received Inhibitors,Modulators,Libraries the sevoflurane at postnatal day 6 or from P6 to P8. Inhibitors,Modulators,Libraries The mice received anesthetic sevoflurane plus 60% oxygen as performed in our previous studies. The 60% oxygen maintains sufficient partial pressure of oxygen levels in the mice during anesthesia as demonstrated in previous studies. The size of the induction chamber in the current study was 20 20 7 centimeters. The induction flow rate was 2 liters per minute for the first three minutes and then one liter per minute. Control group received 60% oxygen at an identical flow rate in simi lar chambers.

The anesthetic and oxygen concentrations were measured continuously by a gas analyzer. The temperature of the anesthetizing chamber was controlled by the DC Temperature Control System, which is a feedback based Imatinib Mesylate Bcr-Abl system for monitoring and con trolling temperature, to maintain the rectal temperature of the mice as 37 0. 5 C. Previous studies have shown that anesthesia with 3% sevoflurane for two hours did not significantly change the values of pH, partial pres sure of oxygen, or partial pressure of carbon dioxide as compared to the control group.