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Volumn 277, Issue 5329, 1997, Pages 1094-1097

Functional role of high-affinity anandamide transport, as revealed by selective inhibition

Author keywords

[No Author keywords available]

Indexed keywords

ANANDAMIDE;

EID: 0030865871     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5329.1094     Document Type: Article
Times cited : (731)

References (41)
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    • Cortical neurons were prepared in 12-well plates and used after 4 to 6 days in vitro. Incubations were carried out in the presence of forskolin (3 μM) and isobutyl methyl xanthine (1 mM). The cAMP concentrations were measured by radioimmunoassay with a commercial kit (Amersham, Arlington, IL) and following manufacturer's instructions.
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    • The amounts of cAMP in the presence of a concentration of WIN-55212-2 below threshold (1 nM, determined in preliminary experiments) were 96.7 ± 2.5% of forskolin alone and were not significantly affected by 10 μM AM404 (89.8 ± 2.6%), 10 μM AM403 (92.4 ± 2.3%), or 10 μM bromcresol green (92.9 ± 2.3%) (n = 3). In the presence of a concentration of glutamate below threshold (3 μM) (24), cAMP concentrations were 91.6 ± 2% of forskolin alone and were not significantly affected by AM404 (84.4 ± 4.9%), AM403 (89.5 ± 2.4%), or bromcresol green (84.4 ± 3%) (n = 3).
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    • The hot plate test (55.5°C) was carried out on male Swiss mice (25 to 30 g, Nossan, Italy) following standard procedures [F. Porreca, H. L. Mosberg, R. Hurst, V. J. Hruby, T. F. Burks, J. Pharmacol. Exp. Ther. 230, 341 (1994)]. Anandamide and AM404 were dissolved in 0.9% NaCl solution containing 20% dimethyl sulfoxide and injected intravenously at 20 mg/kg and 10 mg/kg, respectively. To determine whether cannabinoid receptors participate in the effect of anandamide, we administered anandamide (20 mg/kg intravenously) or anandamide plus SR141716-A (2 mg/kg, subcutaneously) to two groups of six mice each. In mice that received anandamide alone, latency to jump increased from 21.7 ± 1.5 s to 30.7 ± 0.8 s (P < 0.05, ANOVA) 20 min after injection. In contrast, in mice that received anandamide plus SR141716-A, the latency to jump was not affected (19.6 ± 3.1 s).
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    • note
    • We thank E. di Tomaso and H. Cadas for help and E. Barker, L. Parsons, and P. Schweitzer for critical reading of the manuscript. Supported by the Neuroscience Research Foundation, which receives major support from Novartis.


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