Modulation of [35S]TBPS binding by ligands with preferential affinity for benzodiazepine BZ1 sites in the cerebral cortex of newborn and adult rats.
The present study was designed to compare the allosteric coupling between the Cl- channel of the GABAA receptor and the different benzodiazepine recognition site subtypes (BZ sites) in the cerebral cortex of newborn (5-day-old) and adult rats (90-day-old). To this aim, we reexamined the heterogeneity of cortical GABAA receptors in self- and cross-competition binding experiments using [3H]flunitrazepam and two ligands with higher affinity for benzodiazepine BZ1 sites relative to benzodiazepine BZ2 sites, the triazolopyridazine 3-methyl-6-[3-(trifluoromethyl)phenyl]-1,2,4-triazolo [4,3-b] pyridazine (CL 218,872) and the imidazopyridine N,N,6-trimethyl-2-(4-methylphenyl)-imidazo[1,2-a]-pyridine-3-acetamide hemitartrate (zolpidem). Benzodiazepine BZ1 sites accounted for 52% of the total number of binding sites in adult rats, but were not detected in newborn rats. On the other hand, two classes of benzodiazepine BZ2 sites with high and low affinity for zolpidem were present in newborn and adult rats. These sites were designated as benzodiazepine BZ2H (high affinity for zolpidem, Kd approximately 150 nM) and benzodiazepine BZ2L (low affinity for zolpidem, Kd approximately 3000 nM). High densities of benzodiazepine BZ2H sites were measured in both newborn and adult rats (75% and 41% of the total number of [3H]flunitrazepam binding sites, respectively), whereas benzodiazepine BZ2L sites accounted for 25% and 7% of the total number of cortical sites in neonates and adults, respectively. Flunitrazepam, CL 218,872 and zolpidem inhibited in a concentration-dependent manner the binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS) to the convulsant site of cortical GABAA receptors in newborn and adult rats. The IC50 for flunitrazepam was about 3-fold greater in adults than in neonates. This rightward shift in the concentration-response curve may be due to a decrease with age in the intrinsic efficacy of flunitrazepam. In contrast, CL 218,872 and zolpidem were 4-fold more potent at inhibiting [35S]TBPS binding in adult rats relative to neonates. The different affinities of CL 218,872 and zolpidem for benzodiazepine BZ1 and BZ2 receptors may account, at least in part, for the age-related changes in their inhibitory potencies. These results demonstrate that benzodiazepine BZ2 sites mediate the modulation of [35S]TBPS binding by benzodiazepine recognition site ligands in the cerebral cortex of newborn rats. Further, benzodiazepine BZ2 sites may be involved in the inhibition of [35S]TBPS binding by flunitrazepam, CL 218,872 and zolpidem in the cerebral cortex of adult rats.
Giorgi
O
O
Department of Toxicology, University of Cagliari, Italy.
Lecca
D
D
Cancedda
E
E
Serra
G P
GP
Corda
M G
MG
D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D001665:Binding Sites; D001643:Bridged Bicyclo Compounds / Q000378:metabolism*; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002540:Cerebral Cortex / Q000378:metabolism*; D005260:Female; D005445:Flunitrazepam / Q000494:pharmacology; D008297:Male; D011247:Pregnancy; D011724:Pyridazines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000378:metabolism*; D000077334:Zolpidem
D001643:Bridged Bicyclo Compounds; D019086:Bridged Bicyclo Compounds, Heterocyclic; D011724:Pyridazines; D011725:Pyridines; D011963:Receptors, GABA-A; D005445:Flunitrazepam; C021604:CL 218872; C037476:tert-butylbicyclophosphorothionate; D000077334:Zolpidem