The role of functional postsynaptic NMDA receptors in the central nucleus of the amygdala in opioid dependence.

Activation of ionotropic N-methyl-D-aspartate (NMDA)-type glutamate receptors in limbic system nuclei, such as the central nucleus of the amygdala (CeA), plays an essential role in autonomic, behavioral, and affective processes that are profoundly impacted by exposure to opioids. However, the heterogeneous ultrastructural distribution of the NMDA receptor, its complex pharmacology, and the paucity of genetic models have hampered the development of linkages between functional amygdala NMDA receptors and opioid dependence. To overcome these shortcomings, high-resolution imaging and molecular pharmacology were used to (1) Identify the ultrastructural localization of the essential NMDA-NR1 receptor (NR1) subunit and its relationship to the mu-opioid receptor (microOR), the major cellular target of abused opioids like morphine, in the CeA and (2) Determine the effect of CeA NR1 deletion on the physical, and particularly, psychological aspects of opioid dependence. Combined immunogold and immuoperoxidase electron microscopic analysis showed that NR1 was prominently expressed in postsynaptic (i.e., somata, dendrites) locations of CeA neurons, where they were also frequently colocalized with the microOR. A spatial-temporal deletion of NR1 in postsynaptic sites of CeA neurons was produced by local microinjection of a neurotropic recombinant adeno-associated virus (rAAV), expressing the green fluorescent protein (GFP) reporter and Cre recombinase (rAAV-GFP-Cre), in adult "floxed" NR1 (fNR1) mice. Mice with deletion of NR1 in the CeA showed no obvious impairments in sensory, motor, or nociceptive function. In addition, when administered chronic morphine, these mice also displayed an acute physical withdrawal syndrome precipitated by naloxone. However, opioid-dependent CeA NR1 knockout mice failed to exhibit a conditioned place aversion induced by naloxone-precipitated withdrawal. These results indicate that postsynaptic NMDA receptor activity in central amygdala neurons is required for the expression of a learned affective behavior associated with opioid withdrawal. The neurogenetic dissociation of physical and psychological properties of opioid dependence demonstrates the value of combined ultrastructural analysis and molecular pharmacology in clarifying the neurobiological mechanisms subserving opioid-mediated plasticity. Copyright 2010 Elsevier Inc. All rights reserved.