title: γ-Hydroxybutyric acid induces actions via the GABAB receptor in arousal and motor control-related nuclei: implications for therapeutic actions in behavioral state disorders.
abstract: γ-Hydroxybutyric acid (GHB) is used as an effective therapeutic for reducing the hypersomnolence and cataplexy (loss of motor control) of the sleeping disorder, narcolepsy, with an immediate pharmacologic behavioral action of inducing a natural sleep-like state. Despite its clinical use, few studies have examined the cellular actions of this drug on behavioral state-related neurons. Therefore, we monitored GHB-induced responses using calcium imaging within the laterodorsal tegmentum (LDT) and the dorsal raphe (DR), two pontine nuclei important in state and motor control. In addition, we recorded GHB-induced membrane responses using whole cell, patch clamp electrophysiology of immunohistochemically-identified principal neurons within these nuclei. GHB induced GABAB receptor-mediated rises in calcium in neurons of the LDT and the DR. However, the pattern and amplitude of calcium rises differed greatly between these two nuclei. GHB induced GABAB receptor antagonist-sensitive outward currents/hyperpolarizations in immunohistochemically-identified cholinergic LDT and serotonergic DR neurons. However, GHB had this action in a greater proportion of DR cells than LDT neurons. Further, larger inhibitory currents were induced in DR cells when compared to the amplitude of GHB-induced current in LDT-responding cells. Finally, NCS-382 and HOCPCA, a reported antagonist and agonist specific to activity at the putative GHB receptor, respectively, with no demonstrated binding at the GABAB receptor, failed to block GHB-induced effects or elicit any discernible electrophysiological action when applied alone, indicating a lack of involvement of a GHB receptor in mediating GHB actions. Taken together, our data support the conclusion that GHB may be exerting its actions on state and motor control, in part, via an acutely mediated strong inhibition of serotonergic DR neurons and a more modest inhibitory action on a smaller proportion of LDT cholinergic neurons. Given the roles played by these nuclei, these actions are consistent with acute pharmacologic effects of GHB: hypotonia and promotion of sleep, including presence of REM, a sub-state of sleep. Differences in GHB-mediated calcium suggest differential regulation of calcium-dependent processes, which may also contribute to functioning of the LDT and DR in state and motor control and the therapeutic pharmacologic actions of GHB, which develop following chronic administration. These findings add to knowledge of cellular actions of GHB and it is hoped that, combined with findings from other studies examining GHB neurotransmission, these data can contribute to development of highly targeted therapeutics at the GABAB receptor for management of human disorders presenting with alterations in motor and arousal control.
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health Sciences, Universitsparken 2, University of Copenhagen, Copenhagen 2100, Denmark. Electronic address: kak1@farma.ku.dk.
authors.forename: K A
authors.initials: KA
authors.lastname: Kohlmeier
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health Sciences, Universitsparken 2, University of Copenhagen, Copenhagen 2100, Denmark.
authors.forename: B
authors.initials: B
authors.lastname: Vardar
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health Sciences, Universitsparken 2, University of Copenhagen, Copenhagen 2100, Denmark.
authors.forename: M H
authors.initials: MH
authors.lastname: Christensen
mesh_terms: D000818:Animals; D001143:Arousal / Q000187:drug effects*; D001522:Behavior, Animal / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D059329:Cholinergic Neurons / Q000378:metabolism; D065847:Dorsal Raphe Nucleus / Q000166:cytology; D004594:Electrophysiology; D006885:Hydroxybutyrates / Q000494:pharmacology*; D053444:Inhibitory Postsynaptic Potentials / Q000187:drug effects; D051379:Mice; D009043:Motor Activity / Q000187:drug effects*; D009415:Nerve Net / Q000378:metabolism*; D009434:Neural Pathways / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods; D018080:Receptors, GABA-B / Q000378:metabolism*; D059326:Serotonergic Neurons / Q000378:metabolism; D013681:Tegmentum Mesencephali / Q000166:cytology
chemical_list: D006885:Hydroxybutyrates; D018080:Receptors, GABA-B; C111420:4-hydroxybutyric acid; D002118:Calcium
keywords: GABA; REM sleep; baclofen; motor atonia; narcolepsy