Nicotinic-agonist stimulated (86)Rb(+) efflux and [(3)H]epibatidine binding of mice differing in beta2 genotype.
Nicotinic acetylcholine receptor function and binding was measured in 12 brain regions from mice differing in beta2 subunit expression. Function was measured by on-line detection of (86)Rb(+) efflux stimulated under conditions that measure two pharmacologically distinct nicotinic responses: (1) stimulation with 10 microM nicotine, a response that is relatively sensitive to inhibition by the antagonist, dihydro-beta-erythroidine (DHbetaE); and (2) stimulation with 10 microM epibatidine in the presence of 2 microM DHbetaE, a response that is relatively resistant to inhibition by DHbetaE. Deletion of the beta2 subunit profoundly reduced both DHbetaE-sensitive and -resistant (86)Rb(+) efflux in each brain region and essentially eliminated activity in regions such as cerebral cortex and thalamus. However, residual activity was observed in regions such as olfactory bulbs and inferior colliculus. [(3)H]Epibatidine binding was measured under conditions that allow estimation of both high- and low-affinity sites. High-affinity sites sensitive to inhibition by the nicotinic agonist, cytisine, were virtually eliminated in every region by the beta2 null mutation. In contrast, only a subset of the high-affinity sites insensitive to inhibition by cytisine were eliminated in beta2 null mutants, suggesting receptor heterogeniety. Similarly, low affinity [(3)H]epibatidine binding was heterogeneous in that a fraction of the sites required the beta2 subunit. Many remaining sites were sensitive to inhibition by alpha-bungarotoxin indicating that a subset of the low affinity [(3)H]epibatidine binding are of the alpha7* subtype. Distinct regional variation was observed among the 12 brain regions. These studies confirm important roles for beta2-containing receptors in mediating pharmacologically distinct functions and as components of several identifiable binding sites.
Marks
M J
MJ
Institute for Behavioral Genetics, University of Colorado, Boulder, CO 80309, USA.
Stitzel
J A
JA
Grady
S R
SR
Picciotto
M R
MR
Changeux
J P
JP
Collins
A C
AC
D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism*; D002038:Bungarotoxins / Q000494:pharmacology; D005260:Female; D005838:Genotype; D007700:Kinetics; D008297:Male; D051379:Mice; D018722:Nicotinic Agonists / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D011725:Pyridines / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D012413:Rubidium / Q000378:metabolism*; D012414:Rubidium Radioisotopes; D013268:Stimulation, Chemical; D014701:Veratridine / Q000494:pharmacology
D019086:Bridged Bicyclo Compounds, Heterocyclic; D002038:Bungarotoxins; D018722:Nicotinic Agonists; D011188:Potassium; D011725:Pyridines; D011978:Receptors, Nicotinic; D012413:Rubidium; D012414:Rubidium Radioisotopes; D014701:Veratridine; C082748:epibatidine