title: Inhibition of heat shock protein 90 attenuates adenylate cyclase sensitization after chronic morphine treatment.
abstract: Cellular adaptations to chronic opioid treatment result in enhanced responsiveness of adenylate cyclase and an increase in forskolin- or agonist-stimulated cAMP production. It is, however, not known whether chaperone molecules such as heat shock proteins contribute to this adenylate cyclase sensitization. Here, we report that treatment of cells with geldanamycin, an inhibitor of heat shock protein 90 (Hsp90), led to effective attenuation of morphine-induced adenylate cyclase sensitization. In SK-N-SH human neuroblastoma cells, morphine significantly increased RNA transcript and protein levels of type I adenylate cyclase, leading to sensitization. Whole-genome tiling array analysis revealed that cAMP response element-binding protein, an important mediator for cellular adaptation to morphine, associated with the proximal promoter of Hsp90AB1 not only in SK-N-SH cells but also in rat PC12 and human embryonic kidney cells. Hsp90AB1 transcript and protein levels increased significantly during morphine treatment, and co-application of geldanamycin (0.1-10 nM) effectively suppressed the increase in forskolin-activated adenylate cyclase activation by 56%. Type I adenylate cyclase, but not Hsp90AB1, underwent significant degradation during geldanamycin treatment. These results indicate that Hsp90 is a new pharmacological target for the suppression of adenylate cyclase sensitization induced by chronic morphine treatment.
authors.affiliation: Division of Molecular Pharmacology, Department of Pharmacology, Jichi Medical University, Tochigi 329-0498, Japan. t_koshi@jichi.ac.jp
authors.forename: Taka-aki
authors.initials: TA
authors.lastname: Koshimizu
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Tsuchiya
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Tsuda
authors.forename: Yoko
authors.initials: Y
authors.lastname: Fujiwara
authors.forename: Katsushi
authors.initials: K
authors.lastname: Shibata
authors.forename: Akira
authors.initials: A
authors.lastname: Hirasawa
authors.forename: Gozoh
authors.initials: G
authors.lastname: Tsujimoto
authors.forename: Akio
authors.initials: A
authors.lastname: Fujimura
mesh_terms: D000262:Adenylyl Cyclases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology; D045744:Cell Line, Tumor; D004361:Drug Tolerance; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D009020:Morphine / Q000494:pharmacology*; D009294:Narcotics / Q000494:pharmacology*; D051381:Rats; D017450:Receptors, Opioid, mu / Q000235:genetics
chemical_list: D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; C493165:HSP90AB1 protein, human; D047029:Lactams, Macrocyclic; D009294:Narcotics; D017450:Receptors, Opioid, mu; D009020:Morphine; D000262:Adenylyl Cyclases; C106670:adenylyl cyclase 1; C001277:geldanamycin