Loss of surface N-methyl-d-aspartate receptor proteins in mouse cortical neurones during anaesthesia induced by chloral hydrate in vivo A. LacKamp, G.-C.

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Loss of surface N-methyl-d-aspartate receptor proteins in mouse cortical neurones during anaesthesia induced by chloral hydrate in vivo A. LacKamp, G.-C. Zhang, L.-M. Mao, E.E. Fibuch, J.Q. Wang British Journal of Anaesthesia Volume 102, Issue 4, Pages 515-522 (April 2009) DOI: 10.1093/bja/aep009 Copyright © 2009 British Journal of Anaesthesia Terms and Conditions

Fig 1 Normal distribution of NMDA receptor subunit proteins in the surface and intracellular compartments of mouse cerebral cortical neurones as detected by BS3-cross-linking assays. (a) Representative immunoblots showing BS3-linked (solid arrows) and BS3-unlinked (open arrows) subunit proteins in BS3-treated tissue and control (Con) tissue without BS3 treatment. Note that intracellular α-actinin proteins show no cross-linking with BS3. (b) Incubation of BS3 with cerebral cortical homogenates eliminates the monomer band of NR1. (c) Co-incubation of BS3 and chloral hydrate (5 mM) with cerebral cortical homogenates eliminates the monomer band of NR1. (d) The percentage of NMDA receptor subunit proteins and α-actinin in the intracellular pool. Total and intracellular proteins at the monomeric molecular weight level were measured from control tissue and BS3-treated tissue, respectively. The data are expressed in terms of means (sem) (n=3–6 per group). British Journal of Anaesthesia 2009 102, 515-522DOI: (10.1093/bja/aep009) Copyright © 2009 British Journal of Anaesthesia Terms and Conditions

Fig 2 Effects of anaesthesia on NMDA receptor subunit expression in surface and intracellular pools of mouse cortical neurones. Effects of anaesthesia on NR1 (a), NR2A (c), and NR2B (e) expression in the two different compartments. Effects of anaesthesia on the ratio of surface to intracellular expression of NR1 (b), NR2A (d), and NR2B (f). Note that anaesthesia reduced NR1 and NR2B abundance in the surface pool whereas increased them in the intracellular pool (a and e). Anaesthesia was induced by chloral hydrate (500 mg kg−1, i.p.), and mice were killed 10 min after drug injection. Control mice received a PBS injection. Representative immunoblots are shown left to the quantified data (a, c, and e). The data are expressed in terms of means (sem) (n=4–6 per group). *P<0.05 vs PBS. British Journal of Anaesthesia 2009 102, 515-522DOI: (10.1093/bja/aep009) Copyright © 2009 British Journal of Anaesthesia Terms and Conditions

Fig 3 Time-dependent effects of anaesthesia on the righting reflex behaviour and compartment-specific expression of NMDA receptor subunits in mouse cortical neurones. (a) Effects of anaesthesia on righting reflex scores in mice. Effects of anaesthesia on NR1 (b), NR2A (c), and NR2B (d) expression in the two different compartments at different time points. Anaesthesia was induced by chloral hydrate (500 mg kg−1, i.p.). Mice were scored in righting reflect assessments at indicated times (a) or killed at different time points for protein expression assays (b–d). Control mice received a PBS injection. The data are expressed in means (sem) (n=4–6 per group). *P<0.05 vs PBS. British Journal of Anaesthesia 2009 102, 515-522DOI: (10.1093/bja/aep009) Copyright © 2009 British Journal of Anaesthesia Terms and Conditions

Fig 4 Effects of anaesthesia on NR1 and NR2B subunit expression in surface and intracellular pools of mouse cerebral cortical neurones. Anaesthesia was induced by chloral hydrate (500 mg kg−1, i.p.), and body temperature was maintained at 37°C. Mice were killed 10 min after injection. Control mice received a PBS injection. The quantified data of NR1 and NR2B subunit proteins were analysed from separate experiments [means (sem), n=4–5 per group]. *P<0.05 vs PBS. British Journal of Anaesthesia 2009 102, 515-522DOI: (10.1093/bja/aep009) Copyright © 2009 British Journal of Anaesthesia Terms and Conditions