The ?-aminobutyric acid type A (GABAA) receptor is a member of the Cys-loop family of transmitter-gated ion channels. The receptors respond to synaptically-released or ambient transmitter with a conformational change, resulting in the opening of the gate that allows ion movement through the channel. Potentiation of GABAA receptor activity underlies the anesthetic effects of many intravenous anesthetics, such as etomidate and propofol. Although not in active clinical use, neuroactive steroids are among the most potent and efficacious potentiators of the GABAA receptor. Propofol, etomidate and neuroactive steroids also directly activate the GABAA receptor, although the concentrations for direct activation are typically higher than those that cause potentiation. Our preliminary data demonstrate that some neuroactive steroids strongly potentiate gating by propofol or etomidate. Combination of a low concentration of GABA with a low concentration of steroid results in a remarkable, supra-additive, increase in the extent of potentiation. In tadpole and mouse behavioral assays, the presence of a low, subthreshold concentration of a potentiating steroid results in a leftward shift in the dose-response relationship for loss of righting whereas in a cel line the presence of a steroid does not affect etomidate-elicited suppression of cortisol release. The overall goal of the present work is to determine the mechanism of interactions of steroid analogues and intravenous anesthetics on GABAA receptors, and to explore the clinical significance of the findings. We will: i) examine the synergistic effects of steroids with allosterc activators on recombinant synaptic and extrasynaptic GABA expressed in heterologous expression systems; ii) examine the behavioral consequences of synergistic effects of steroids and intravenous anesthetics in tadpole and mouse behavioral assays; and iii) probe whether reduced doses of anesthetics required to produce sedation in the presence of steroids lead to reduced off-target effects. The completion of these aims will increase our understanding of how the GABAA receptor functions in health and disease, and lay a foundation for future development of novel therapeutic approaches.

Public Health Relevance

Potentiation of ?-aminobutyric type A (GABAA) receptors underlies the anesthetic effects of intravenous anesthetics such as etomidate, propofol and barbiturates. This project examines the effect of steroids and steroid analogues on the dosage requirements for intravenous anesthetics to potentiate GABAA receptors and to produce sedation without affecting adverse side effects in animals.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM108580-04
Application #
9319277
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Cole, Alison E
Project Start
2014-08-01
Project End
2019-07-31
Budget Start
2017-08-01
Budget End
2019-07-31
Support Year
4
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Washington University
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Cheng, Wayland W L; Chen, Zi-Wei; Bracamontes, John R et al. (2018) Mapping two neurosteroid-modulatory sites in the prototypic pentameric ligand-gated ion channel GLIC. J Biol Chem 293:3013-3027
Cao, Lily Q; Montana, Michael C; Germann, Allison L et al. (2018) Enhanced GABAergic actions resulting from the coapplication of the steroid 3?-hydroxy-5?-pregnane-11,20-dione (alfaxalone) with propofol or diazepam. Sci Rep 8:10341
Germann, Allison L; Shin, Daniel J; Kuhrau, Christina R et al. (2018) High Constitutive Activity Accounts for the Combination of Enhanced Direct Activation and Reduced Potentiation in Mutated GABAA Receptors. Mol Pharmacol 93:468-476
Shin, Daniel J; Germann, Allison L; Johnson, Alexander D et al. (2018) Propofol Is an Allosteric Agonist with Multiple Binding Sites on Concatemeric Ternary GABAA Receptors. Mol Pharmacol 93:178-189
Sun, Min-Yu; Shu, Hong-Jin; Benz, Ann et al. (2018) Chemogenetic Isolation Reveals Synaptic Contribution of ? GABAA Receptors in Mouse Dentate Granule Neurons. J Neurosci 38:8128-8145
Akk, Gustav; Shin, Daniel J; Germann, Allison L et al. (2018) GABA Type A Receptor Activation in the Allosteric Coagonist Model Framework: Relationship between EC50 and Basal Activity. Mol Pharmacol 93:90-100
Shin, Daniel J; Germann, Allison L; Steinbach, Joe Henry et al. (2017) The Actions of Drug Combinations on the GABAA Receptor Manifest as Curvilinear Isoboles of Additivity. Mol Pharmacol 92:556-563
Bracamontes, John R; Akk, Gustav; Steinbach, Joe Henry (2016) Introduced Amino Terminal Epitopes Can Reduce Surface Expression of Neuronal Nicotinic Receptors. PLoS One 11:e0151071
Eaton, Megan M; Germann, Allison L; Arora, Ruby et al. (2016) Multiple Non-Equivalent Interfaces Mediate Direct Activation of GABAA Receptors by Propofol. Curr Neuropharmacol 14:772-80
Germann, Allison L; Shin, Daniel J; Manion, Brad D et al. (2016) Activation and modulation of recombinant glycine and GABAA receptors by 4-halogenated analogues of propofol. Br J Pharmacol 173:3110-3120

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