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Mice were injected with morphine (10 mg/kg sc). After 30 min or 2 hours, wild-type mice were killed and blood was collected in vials containing sodium fluoride and potassium oxalate. Morphine concentrations in blood samples pooled from three mice per sample were 1500 ng/ml after 30 min and 83 ng/ml after 2 hours, as measured by mass spectroscopy analysis (Occupational Testing Division, LabCorp Inc., Research Triangle Park, NC). In similar experiments, βarr2-KO mice had a cocentration of 93 ng/ml in the blood after 2 hours.
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Naloxone (2.5 mg/kg sc), which immediately reverses the effects of opiates, was given 30 min after morphine (10 mg/kg). Naltrindole [P. S. Portoghese, M. Sultana, A. E. Takemori. J. Med. Chem. 88, 1547 (1990)] was given 20 min before morphine, and nor-binaltorphimine [A. E. Takemori, B. Y. Ho, J. S. Naeseth, P. S. Portoghese, J. Phamacol. Exp. Ther. 246, 255 (1988)] was given 1 hour before morphine [H. W. Matthes et al., J. Neurosci. 18, 7285 (1998)].
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24
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Naloxone (2.5 mg/kg sc), which immediately reverses the effects of opiates, was given 30 min after morphine (10 mg/kg). Naltrindole [P. S. Portoghese, M. Sultana, A. E. Takemori. J. Med. Chem. 88, 1547 (1990)] was given 20 min before morphine, and nor-binaltorphimine [A. E. Takemori, B. Y. Ho, J. S. Naeseth, P. S. Portoghese, J. Phamacol. Exp. Ther. 246, 255 (1988)] was given 1 hour before morphine [H. W. Matthes et al., J. Neurosci. 18, 7285 (1998)].
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Naloxone (2.5 mg/kg sc), which immediately reverses the effects of opiates, was given 30 min after morphine (10 mg/kg). Naltrindole [P. S. Portoghese, M. Sultana, A. E. Takemori. J. Med. Chem. 88, 1547 (1990)] was given 20 min before morphine, and nor-binaltorphimine [A. E. Takemori, B. Y. Ho, J. S. Naeseth, P. S. Portoghese, J. Phamacol. Exp. Ther. 246, 255 (1988)] was given 1 hour before morphine [H. W. Matthes et al., J. Neurosci. 18, 7285 (1998)].
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γ-S binding assays. For both binding assays, reactions were terminated by rapid filtration over GF/B filters (Brandel Inc., Caithersburg, MD) using a Brandel cell harvester. Filters were washed three times with ice-cold 10 mM tris-HCl (pH 7.4) and then counted in a liquid scintillation counter.
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We thank B. H. Koller for homologous recombinant mouse embryonic stem cells and chimeric mice, M. R. Capecchi for plasmid plC19R/MCl-TK, and R. Hen for plasmid pD383. R.R.G. is a visiting scientist from the Institute of Pharmacology, Russian Academy of Medical Sciences, Moscow. Supported in part by NIH grants NS 19576 (M.G.C.) and HL16037, and by a cardiovascular unrestricted grant from Bristol-Myers Squibb (R.J.L). R.J.L. and M.G.C. are Investigators of the Howard Hughes Medical Institute.
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