title: GABAAreceptor subtypes in the mouse brain: Regional mapping and diazepam receptor occupancy by in vivo [18F]flumazenil PET.
abstract: Classical benzodiazepines, which are widely used as sedatives, anxiolytics and anticonvulsants, exert their therapeutic effects through interactions with heteropentameric GABAAreceptors composed of two α, two β and one γ2 subunit. Their high affinity binding site is located at the interface between the γ2 and the adjacent α subunit. The α-subunit gene family consists of six members and receptors can be homomeric or mixed with respect to the α-subunits. Previous work has suggested that benzodiazepine binding site ligands with selectivity for individual GABAAreceptor subtypes, as defined by the benzodiazepine-binding α subunit, may have fewer side effects and may even be effective in diseases, such as schizophrenia, autism or chronic pain, that do not respond well to classical benzodiazepines. The distributions of the individual α subunits across the CNS have been extensively characterized. However, as GABAAreceptors may contain two different α subunits, the distribution of the subunits does not necessarily reflect the distribution of receptor subtypes with respect to benzodiazepine pharmacology. In the present study, we have used in vivo [18F]flumazenil PET and in vitro [3H]flumazenil autoradiography in combination with GABAAreceptor point-mutated mice to characterize the distribution of the two most prevalent GABAAreceptor subtypes (α1 and α2) throughout the mouse brain. The results were in agreement with published in vitro data. High levels of α2-containing receptors were found in brain regions of the neuronal network of anxiety. The α1/α2 subunit combinations were predictable from the individual subunit levels. In additional experiments, we explored in vivo [18F]flumazenil PET to determine the degree of receptor occupancy at GABAAreceptor subtypes following oral administration of diazepam. The dose to occupy 50% of sensitive receptors, independent of the receptor subtype(s), was 1-2mg/kg, in agreement with published data from ex vivo studies with wild type mice. In conclusion, we have resolved the quantitative distribution of α1- and α2-containing homomeric and mixed GABAAreceptors in vivo at the millimeter scale and demonstrate that the regional drug receptor occupancy in vivo at these GABAAreceptor subtypes can be determined by [18F]flumazenil PET. Such information should be valuable for drug development programs aiming for subtype-selective benzodiazepine site ligands for new therapeutic indications.
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences (D-CHAB), ETH Zurich, Vladimir-Prelog-Weg 4, CH-8093 Zurich, Switzerland.
authors.forename: Adrienne
authors.initials: A
authors.lastname: Müller Herde
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Benke
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
authors.forename: William T
authors.initials: WT
authors.lastname: Ralvenius
authors.affiliation: Department of Nuclear Medicine, University Hospital Zurich, Rämistrasse 100, CH-8091 Zurich, Switzerland.
authors.forename: Linjing
authors.initials: L
authors.lastname: Mu
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences (D-CHAB), ETH Zurich, Vladimir-Prelog-Weg 4, CH-8093 Zurich, Switzerland.
authors.forename: Roger
authors.initials: R
authors.lastname: Schibli
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences (D-CHAB), ETH Zurich, Vladimir-Prelog-Weg 4, CH-8093 Zurich, Switzerland; Institute of Pharmacology and Toxicology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
authors.forename: Hanns Ulrich
authors.initials: HU
authors.lastname: Zeilhofer
authors.affiliation: Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences (D-CHAB), ETH Zurich, Vladimir-Prelog-Weg 4, CH-8093 Zurich, Switzerland. Electronic address: Stefanie.Kraemer@pharma.ethz.ch.
authors.forename: Stefanie D
authors.initials: SD
authors.lastname: Krämer
mesh_terms: D000818:Animals; D001345:Autoradiography; D001921:Brain / Q000378:metabolism*; D003975:Diazepam / Q000494:pharmacology; D005442:Flumazenil; D005462:Fluorine Radioisotopes; D018757:GABA Modulators / Q000494:pharmacology; D051379:Mice; D008817:Mice, Mutant Strains; D059906:Neuroimaging / Q000379:methods*; D049268:Positron-Emission Tomography / Q000379:methods*; D019275:Radiopharmaceuticals; D011963:Receptors, GABA-A / Q000032:analysis / Q000096:biosynthesis*
chemical_list: D005462:Fluorine Radioisotopes; D018757:GABA Modulators; D019275:Radiopharmaceuticals; D011963:Receptors, GABA-A; D005442:Flumazenil; D003975:Diazepam
keywords: Alpha subunit; Benzodiazepine; GABA(A) receptor; Receptor occupancy; Selective