Kenjiro Tanaka1, Takahiro Shimizu2, Youichirou Higashi3, Kumiko Nakamura1, Keisuke Taniuchi1, Fotios Dimitriadis4, Shogo Shimizu5, Kunihiko Yokotani1, Motoaki Saito1. 1. Department of Pharmacology, Kochi University School of Medicine, Nankoku, Kochi 783-8505, Japan. 2. Department of Pharmacology, Kochi University School of Medicine, Nankoku, Kochi 783-8505, Japan. Electronic address: shimizu@kochi-u.ac.jp. 3. Department of Neurosurgery, Kochi University School of Medicine, Nankoku, Kochi 783-8505, Japan. 4. B' Urologic Department, Papageorgiou General Hospital, Aristotle University School of Medicine, Thessaloniki, Greece. 5. Division of Molecular Pharmacology, Tottori University School of Medicine, Yonago, Tottori 683-8503, Japan.
Abstract
AIMS: Cyclooxygenase (COX) can be activated by nitric oxide-induced (NO-induced) conversion of cysteine thiol group of COX into S-nitrosothiol. We previously reported the involvement of brain COX/NO synthase (NOS) in centrally administered bombesin-, a stress-related neuropeptide, induced secretion of rat adrenal noradrenaline and adrenaline. To examine a possible involvement of the NO-induced modification of COX in bombesin-induced response, we investigated whether bombesin induces close proximity of COX-1 and neuronal NOS (nNOS) or S-nitroso-cysteine in pre-sympathetic spinally projecting neurons in the rat hypothalamic paraventricular nucleus (PVN), a regulatory center of adrenomedullary outflow. MAIN METHODS: In twelve-week-old male Wistar rats, pre-sympathetic spinally projecting neurons in the PVN were labeled with a retrograde tracer Fluoro-Gold (FG). After intracerebroventricular administration of bombesin, we performed double immunohistochemical analysis for Fos and COX-1 or nNOS in FG-labeled PVN neurons. We also performed a fluorescent in situ proximity ligation assay (PLA) for visualizing of close proximity (<40 nm) of COX-1 with nNOS or S-nitroso-cysteine. KEY FINDINGS: Bombesin significantly increased the number of Fos-immunoreactive cells in FG-labeled PVN neurons with COX-1 or nNOS immunoreactivity. 7-Nitroindazole, a selective nNOS inhibitor, abolished Fos-immunoreactivity induced by bombesin in COX-1-immunoreactive FG-labeled PVN neurons. Bombesin also induced PLA-positive signals indicating close proximity of COX-1/nNOS and COX-1/S-nitroso-cysteine in FG-labeled PVN neurons. SIGNIFICANCE: Centrally administered bombesin possibly induces S-nitrosylation of COX-1 through close proximity of COX-1 and nNOS in pre-sympathetic spinally projecting PVN neurons, thereby activating COX-1 during the bombesin-induced activation of central adrenomedullary outflow in the rat.
AIMS: Cyclooxygenase (COX) can be activated by nitric oxide-induced (NO-induced) conversion of cysteine thiol group of COX into S-nitrosothiol. We previously reported the involvement of brain COX/NO synthase (NOS) in centrally administered bombesin-, a stress-related neuropeptide, induced secretion of rat adrenal noradrenaline and adrenaline. To examine a possible involvement of the NO-induced modification of COX in bombesin-induced response, we investigated whether bombesin induces close proximity of COX-1 and neuronal NOS (nNOS) or S-nitroso-cysteine in pre-sympathetic spinally projecting neurons in the rat hypothalamic paraventricular nucleus (PVN), a regulatory center of adrenomedullary outflow. MAIN METHODS: In twelve-week-old male Wistar rats, pre-sympathetic spinally projecting neurons in the PVN were labeled with a retrograde tracer Fluoro-Gold (FG). After intracerebroventricular administration of bombesin, we performed double immunohistochemical analysis for Fos and COX-1 or nNOS in FG-labeled PVN neurons. We also performed a fluorescent in situ proximity ligation assay (PLA) for visualizing of close proximity (<40 nm) of COX-1 with nNOS or S-nitroso-cysteine. KEY FINDINGS: Bombesin significantly increased the number of Fos-immunoreactive cells in FG-labeled PVN neurons with COX-1 or nNOS immunoreactivity. 7-Nitroindazole, a selective nNOS inhibitor, abolished Fos-immunoreactivity induced by bombesin in COX-1-immunoreactive FG-labeled PVN neurons. Bombesin also induced PLA-positive signals indicating close proximity of COX-1/nNOS and COX-1/S-nitroso-cysteine in FG-labeled PVN neurons. SIGNIFICANCE: Centrally administered bombesin possibly induces S-nitrosylation of COX-1 through close proximity of COX-1 and nNOS in pre-sympathetic spinally projecting PVN neurons, thereby activating COX-1 during the bombesin-induced activation of central adrenomedullary outflow in the rat.
Authors: Xianli Shen; Miguel A Burguillos; Ahmed M Osman; Jeroen Frijhoff; Alejandro Carrillo-Jiménez; Sachie Kanatani; Martin Augsten; Dalel Saidi; Johanna Rodhe; Edel Kavanagh; Anthony Rongvaux; Vilma Rraklli; Ulrika Nyman; Johan Holmberg; Arne Östman; Richard A Flavell; Antonio Barragan; Jose Luis Venero; Klas Blomgren; Bertrand Joseph Journal: Nat Immunol Date: 2016-09-12 Impact factor: 31.250