Inhalational Anesthetics Disrupt Postsynaptic Density Protein-95, Drosophila Disc Large Tumor Suppressor, and Zonula Occludens-1 Domain Protein Interactions Critical to Action of Several Excitatory Receptor Channels Related to Anesthesia

Abstract

Background We have shown previously that inhaled anesthetics disrupt the interaction between the second postsynaptic density protein-95, Drosophila disc large tumor suppressor, and zonula occludens-1 (PDZ) domain of postsynaptic density protein-95 (PSD-95) and the C-terminus of N-methyl-D-aspartate receptor subunits NR2A and NR2B. Our data indicate that PDZ domains may serve as a molecular target for inhaled anesthetics. However, the underlying molecular mechanisms remain to be illustrated. Methods Glutathione S-transferase pull-down assay, co-immunoprecipitation and yeast two-hybrid analysis were used to assess PDZ domain-mediated protein-protein interactions in different conditions. Nuclear magnetic resonance spectroscopy was used to investigate isoflurane-induced chemical shift changes in the PDZ1–3 domains of PSD-95. A surface plasmon resonance-based BIAcore assay was used to examine the ability of isoflurane to inhibit the PDZ domain-mediated protein-protein interactions in real time. Results Halothane and isoflurane dose dependently inhibited PDZ domain-mediated interactions between PSD-95 and Shaker-type potassium channel Kv1.4 and between α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor subunit GluA2 and its interacting proteins— glutamate receptor interacting protein or protein interacting with c kinase 1. However, halothane and isoflurane had no effect on PDZ domain-mediated interactions between γ-aminobutyric acid, type B receptor and its interacting proteins. The inhaled anesthetic isoflurane mostly affected the residues close to or in the peptide binding groove of PSD-95 PDZ1 and PDZ2 (especially PDZ2), while barely affecting the peptide binding groove of PSD-95 PDZ3. Conclusion These results suggest that inhaled anesthetics interfere with PDZ domain-mediated protein-protein interactions at several receptors important to neuronal excitation, anesthesia and pain processing.

Sensitivity to isoflurane anesthesia increases in autism spectrum disorder Shank3 +/∆c mutant mouse model

“…20–22 The isoflurane concentration was reduced by 0.1% for 20 min if the animal did not right itself, and the righting reflex was subsequently retested. RREC 50 was calculated for each mouse as the mean value of the anesthetic concentrations that just permitted and just prevented the righting reflex.…”

“…9,10 Shank3 protein is a molecular scaffolding protein essential for synapse formation and for mediating N-methyl-D-aspartate receptor (NMDAR)- and metabotropic glutamate receptor (mGluR)-induced excitatory synaptic transmission, 11–14 potential sites of anesthetic action. 15–20 We have shown previously that inhalational anesthetic agents can disrupt the interaction of other scaffolding proteins (PSD93 and PSD95) with NMDA and AMPA receptors, resulting in a reduction of the minimum alveolar concentration (MAC) required for anesthesia 21,22 . Shank binds to PSD95-associated protein GKAP and assembles into a complex of Shank/GKAP/PSD95, coupling NMDAR-PSD95 complexes to regulators of the actin cytoskeleton 23 .…”

Molecular Mechanisms of Anesthetic Neurotoxicity: A Review of the Current Literature

“…These findings revealed the PDZ domain as a new molecular target for inhalational anesthetics. Using a surface plasmon resonance-based BIAcore assay, binding of PSD-95 PDZ2 to NR2B C-terminus was shown to be disrupted by isoflurane in real time 86 . These results demonstrated that anesthetics can disrupt the PDZ domain-mediated protein interactions between PSD-95 and the NMDA receptor and suggest downstream effects on synaptic plasticity.…”

“…These results demonstrated that anesthetics can disrupt the PDZ domain-mediated protein interactions between PSD-95 and the NMDA receptor and suggest downstream effects on synaptic plasticity. Given the critical role of PSD-95 and NMDA receptors in synapse development, it has been proposed that anesthetics such as isoflurane and sevoflurane might interfere with brain development by disrupting the interactions between these proteins 86 .…”

5-HT_1A Receptors Mediate Analgesia Induced by Emulsified Sevoflurane in Thermal Nociception but Have Little Effect on Chemical Nociception

“…Various in vitro studies have found that the enhancement of inhibitory neurotransmitter function and inhibition of excitatory neurotransmitter function to be plausible mechanisms underlying the action of inhalation anesthetics. Analgesic effects of anesthetics may be related to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor [4,5,6], benzodiazepine receptor [7], N-methyl-D-Aspartate receptor [8,9,10], glycine receptor [11,12,13,14], nerve nicotinic receptors [15], and alpha 2 receptors [16] on the spinal cord. Morphological and molecular biology research has indicated that the 5-serotonin receptor 1A (5-HT 1A R) is widely distributed in the nervous system.…”