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capacity and curiosity. Mice were placed individually
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equal acrylic compartments, one dark and one light, illu-
minated by a 60 W bulb lamp and separated by a divider
with a 10 · 3-cm opening at floor level. Each mouse was
tested by placing it in the center of the lighted area, fac-
ing away from the dark one, and allowing it to explore
the novel environment for 5 min. The number of trans-
fers from one compartment to the other and the time
spent in the illuminated side were measured. This test
exploited the conflict between the animal’s tendency to
explore a new environment and its fear of bright light.
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6.3.5. Pentylenetetrazole (PTZ)-induced seizure. PTZ
(90 mg/kg sc) was injected 30 min after the administration
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6.3.6. Ethanol-induced sleeping time test. Ethanol (4 g/kg
ip) was injected 30 min after drug administration. The
duration of a loss of the righting reflex was measured
as the sleep time. If the mice slept more than 210 min,
the end-point was recorded as 210 min.
Drugs. Diazepam (Valium 10—Roche), Flumazenil
(Roche), Pentylenetetrazole (PTZ) (Sigma), and Zolpi-
dem (Tocris) were the drugs used. All drugs except
PTZ were suspended in 1% carboxymethylcellulose sodi-
um salt and sonicated immediately before use. PTZ was
dissolved in isotonic (NaCl 0.9%) saline solution and
injected sc. All benzodiazepine receptor ligands were
administered by the po route, except for flumazenil
which was administered ip. Drug concentrations were
prepared in such a way that the necessary dose could
be administered in a 10 ml/kg volume of carboxymethyl-
cellulose (CMC) 1% by the po, ip or sc routes.
Statistical analysis. Results are given as means SEM.
Statistical analysis was performed by means of ANO-
VA, followed by Scheffe’s post-hoc test. Student’s two-
tailed t-test was used to verify significance between
two means. Data were analyzed using a computer pro-
gram (Number Cruncher Statistical System, Version
5.03 9/92). For percentage values, chi-square analysis
was used in accordance with Tallarida & Murray. P
values of less than 0.05 were considered significant.
17. McKay, P. F.; Foster, K. L.; Mason, D.; Cummings, R.;
Garcia, M.; Williams, L. S.; Grey, C.; McCane, S.; He, X.;
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455–462.
18. Costanzo, A.; Guerrini, G.; Ciciani, G.; Bruni, F.; Selleri,
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Aiello, P.; Ipponi, A. J. Med. Chem. 1999, 42, 2218–2226.
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References and notes
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