G-protein receptor kinase 3 (GRK3) influences opioid analgesic tolerance but not opioid withdrawal.

1. Tolerance to opioids frequently follows repeated drug administration and affects the clinical utility of these analgesics. Studies in simple cellular systems have demonstrated that prolonged activation of opioid receptors produces homologous receptor desensitization by G-protein receptor kinase mediated receptor phosphorylation and subsequent beta-arrestin binding. To define the role of this regulatory mechanism in the control of the electrophysiological and behavioral responses to opioids, we used mice having a targeted disruption of the G-protein receptor kinase 3 (GRK3) gene. 2. Mice lacking GRK3 did not differ from wild-type littermates neither in their response latencies to noxious stimuli on the hot-plate test nor in their acute antinociceptive responses to fentanyl or morphine. 3. Tolerance to the electrophysiological response to the opioid fentanyl, measured in vitro in the hippocampus, was blocked by GRK3 deletion. In addition, tolerance to the antinociceptive effects of fentanyl was significantly reduced in GRK3 knockouts compared to wild-type littermate controls. 4. Tolerance to the antinociceptive effects of morphine was not affected by GRK3 deletion although morphine tolerance in hippocampal slices from GRK3 knockout mice was significantly inhibited. Tolerance developed more slowly in vitro to morphine than fentanyl supporting previous work in in vitro systems showing a correlation between agonist efficacy and GRK3-mediated desensitization. 5. The results of these studies suggest that GRK3-mediated mechanisms are important components of both electrophysiologic and behavioral opioid tolerance. Fentanyl, a high efficacy opioid, more effectively produced GRK3-dependent effects than morphine, a low efficacy agonist.

Effect of laryngeal mask airway on esophageal motility during general anesthesia.

STUDY OBJECTIVES: To determine whether the Laryngeal Mask Airway (LMA) triggers the pharyngo-esophago-gastric reflex during general anesthesia by comparing the esophageal motility of patients with the LMA and endotracheal tube (ETT) in place. DESIGN: Randomized clinical trial. SETTING: Operating room and recovery room of a tertiary-care referral hospital. PATIENTS: 50 adult ASA physical status I and II patients scheduled for elective orthopedic surgery. INTERVENTIONS: All patients received a standardized general anesthetic technique, then were allocated randomly to the LMA (n = 30) or ETT (n = 20) groups. MEASUREMENTS AND MAIN RESULTS: The esophageal manometric inputs were recorded continuously using an ambulatory esophageal manometric recorder and divided into five perioperative phases (preanesthesia, induction, surgery, LMA, or ETT rejection and arousal phase). The LMA or ETT was removed at the end of the surgery, when the patient was awake. An awake state was defined as the presence of the following clinical signs: swallowing, bucking, struggling, straining, and restlessness. The esophageal peristaltic wave percent and esophageal contraction frequency were significantly decreased during induction, surgery, and the LMA or ETT rejection and arousal phases compared with the preanesthetic phases in both the LMA and ETT groups. However, there were no significant group differences in any corresponding perioperative phases. CONCLUSION: During the general anesthetic period before the arousal phase in this study, a LMA does not provoke significantly different esophageal peristalsis compared with an ETT. Thus, the LMA is unlikely to stimulate the pharyngo-esophago-gastric reflex during that period.

Regulation of cyclic AMP-dependent response element-binding protein (CREB) by the nociceptin/orphanin FQ in human dopaminergic SH-SY5Y cells.

Nociceptin/orphanin FQ (N/OFQ), an endogenous ligand for opioid receptor-like (ORL1) receptor, transduces signaling cascades implicated in MAPK, PKC, PLC, and calcium, etc. This study was designed to investigate the intracellular signaling mechanism of N/OFQ in human dopaminergic neuroblastoma SH-SY5Y cells. N/OFQ rapidly induced the phosphorylation of CREB, which was significantly suppressed by pretreatment of PKA inhibitor, but not by MAPK inhibitors. It also time-dependently increased the phosphorylation of MAPK, which was proven as ERKs, whereas it did not affect the PI3K activity. Interestingly, KT5720, a specific inhibitor of PKA, markedly suppressed the phosphorylation of MAPK by N/OFQ in SH-SY5Y cells. Furthermore, BAPTA-AM, an intracellular chelator of Ca(2+), completely abolished the phosphorylation of CREB as well as MAPK in N/OFQ-treated SH-SY5Y cells. Taken together, these results suggest that N/OFQ independently induces the activation of CREB prior to MAPK phosphorylation, which was also modulated by PKA. Furthermore, Ca(2+)-related signaling implicates in the phosphorylation processes of CREB and MAPK simultaneously.