title: Sympathetic activity induced by naloxone-precipitated morphine withdrawal is blocked in genetically engineered mice lacking functional CRF1 receptor.
abstract: There is large body evidence indicating that stress can lead to cardiovascular disease. However, the exact brain areas and the mechanisms involved remain to be revealed. Here, we performed a series of experiments to characterize the role of CRF1 receptor (CRF1R) in the stress response induced by naloxone-precipitated morphine withdrawal. The experiments were performed in the hypothalamic paraventricular nucleus (PVN) ventrolateral medulla (VLM), brain regions involved in the regulation of cardiovascular activity, and in the right ventricle by using genetically engineered mice lacking functional CRF1R levels (KO). Mice were treated with increasing doses of morphine and withdrawal was precipitated by naloxone administration. Noradrenaline (NA) turnover, c-Fos, expression, PKA and TH phosphorylated at serine 40, was evaluated by high-performance liquid chromatography (HPLC), immunohistochemistry and immunoblotting. Morphine withdrawal induced an enhancement of NA turnover in PVN in parallel with an increase in TH neurons expressing c-Fos in VLM in wild-type mice. In addition we have demonstrated an increase in NA turnover, TH phosphorylated at serine 40 and PKA levels in heart. The main finding of the present study was that NA turnover, TH positive neurons that express c-Fos, TH phosphorylated at serine 40 and PKA expression observed during morphine withdrawal were significantly inhibited in CRF1R KO mice. Our results demonstrate that CRF/CRF1R activation may contribute to the adaptive changes induced by naloxone-precipitated withdrawal in the heart and in the brain areas which modulate the cardiac sympathetic function and suggest that CRF/CRF1R pathways could be contributing to cardiovascular disease associated to opioid addiction.
authors.affiliation: Department of Pharmacology, School of Medicine, University of Murcia, Spain.
authors.forename: Juan-Antonio
authors.initials: JA
authors.lastname: García-Carmona
authors.affiliation: Department of Pharmacology, School of Medicine, University of Murcia, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Martínez-Laorden
authors.affiliation: Department of Pharmacology, School of Medicine, University of Murcia, Spain.
authors.forename: María-Victoria
authors.initials: MV
authors.lastname: Milanés
authors.affiliation: Department of Pharmacology, School of Medicine, University of Murcia, Spain.
authors.forename: María-Luisa
authors.initials: ML
authors.lastname: Laorden
mesh_terms: D000818:Animals; D001835:Body Weight; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D006321:Heart / Q000033:anatomy & histology; D007031:Hypothalamus / Q000378:metabolism*; D008297:Male; D018345:Mice, Knockout; D009021:Morphine Dependence / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D009270:Naloxone; D009292:Narcotic Antagonists; D009474:Neurons / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D018019:Receptors, Corticotropin-Releasing Hormone / Q000172:deficiency* / Q000235:genetics; D013375:Substance Withdrawal Syndrome / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D000099052:CRF Receptor, Type 1
chemical_list: D009292:Narcotic Antagonists; D016760:Proto-Oncogene Proteins c-fos; D018019:Receptors, Corticotropin-Releasing Hormone; D009270:Naloxone; D000099052:CRF Receptor, Type 1; D014446:Tyrosine 3-Monooxygenase; D017868:Cyclic AMP-Dependent Protein Kinases; D009638:Norepinephrine
keywords: CRF1R knockout mice; Heart; Hypothalamic paraventricular nucleus; Naloxone-precipitated morphine withdrawal; Ventrolateral medulla