? This revised application is being submitted to provide Dr. Michael Glass, with four years of mentored training in molecular neuropharmacological and stereological methods to study drug abuse under the primary sponsorship of Dr Charles Inturissi and co-sponsorship of Dr. Virginia Pickel. This project will allow the candidate to engage in full-time research which will be facilitated by his appointment to tenure-track assistant professor at the Weill Medical College of Cornell University. The projects in this proposal will address the role of amygdala gene expression of the essential NMDA-NR1 (NR1) receptor subunit in the development and expression of opiate dependence by creating spatial-temporal knockouts using a specific and versatile form of Cre-loxP technology. This Cre-loxP system has been successfully used to achieve conditional NR1 gene knockout in spinal cord (i.e. spatial knockout) in adult animals (i.e. temporal knockout) with changes in injury-induced pain. The viability of this conditional gene knockout approach to achieve significant reductions of NR1 gene expression in a specific brain region of adult animals, and its consequences for behavior, neural activation, and protein targeting in dendritic spines has never been determined. This proposal will integrate innovative methods for conditional gene knockout and high resolution quantitative electron microscopic methods along with established behavioral paradigms to achieve the following aims: 1). produce NR1 knockout limited to the amygdala of adult mice, 2). examine the effects of conditional NR1 knockout in the amygdala on the development and expression of morphine dependence and tolerance, 3). determine the effects of conditional NR1 knockout in the amygdala on neural activation induced by opiate withdrawal, 4). examine the effects of amygdala NR1 knockout on the cellular targeting of proteins that are structurally and functionally linked to NR1, including the NMDA-R2 (NR2) subunit and neuronal nitric oxide synthase (nNOS) in dendritic spines. These studies will provide important new information on the role of amygdala NR1 subunit gene expression in the development of opiate dependence, and provide novel insights into the mechanisms mediating neural plasticity induced by chronic opiate use with implications for maintenance of drug use and relapse. ? ? ? ?

National Institute of Health (NIH)
National Institute on Drug Abuse (NIDA)
Research Scientist Development Award - Research & Training (K01)
Project #
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Study Section
Human Development Research Subcommittee (NIDA)
Program Officer
Satterlee, John S
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Weill Medical College of Cornell University
Schools of Medicine
New York
United States
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Beckerman, Marc A; Van Kempen, Tracey A; Justice, Nicholas J et al. (2013) Corticotropin-releasing factor in the mouse central nucleus of the amygdala: ultrastructural distribution in NMDA-NR1 receptor subunit expressing neurons as well as projection neurons to the bed nucleus of the stria terminalis. Exp Neurol 239:120-32
Beckerman, Marc A; Glass, Michael J (2012) The NMDA-NR1 receptor subunit and the mu-opioid receptor are expressed in somatodendritic compartments of central nucleus of the amygdala neurons projecting to the bed nucleus of the stria terminalis. Exp Neurol 234:112-26
Glass, Michael J (2010) The role of functional postsynaptic NMDA receptors in the central nucleus of the amygdala in opioid dependence. Vitam Horm 82:145-66
Jaferi, A; Pickel, V M (2009) Mu-opioid and corticotropin-releasing-factor receptors show largely postsynaptic co-expression, and separate presynaptic distributions, in the mouse central amygdala and bed nucleus of the stria terminalis. Neuroscience 159:526-39
Glass, M J; Vanyo, L; Quimson, L et al. (2009) Ultrastructural relationship between N-methyl-D-aspartate-NR1 receptor subunit and mu-opioid receptor in the mouse central nucleus of the amygdala. Neuroscience 163:857-67
Glass, Michael J; Lane, Diane A; Colago, Eric E O et al. (2008) Chronic administration of morphine is associated with a decrease in surface AMPA GluR1 receptor subunit in dopamine D1 receptor expressing neurons in the shell and non-D1 receptor expressing neurons in the core of the rat nucleus accumbens. Exp Neurol 210:750-61
Glass, Michael J; Hegarty, Deborah M; Oselkin, Martin et al. (2008) Conditional deletion of the NMDA-NR1 receptor subunit gene in the central nucleus of the amygdala inhibits naloxone-induced conditioned place aversion in morphine-dependent mice. Exp Neurol 213:57-70
Glass, Michael J; Kruzich, Paul J; Colago, Eric E O et al. (2005) Increased AMPA GluR1 receptor subunit labeling on the plasma membrane of dendrites in the basolateral amygdala of rats self-administering morphine. Synapse 58:1-12
Glass, Michael J; Kruzich, Paul J; Kreek, Mary Jeanne et al. (2004) Decreased plasma membrane targeting of NMDA-NR1 receptor subunit in dendrites of medial nucleus tractus solitarius neurons in rats self-administering morphine. Synapse 53:191-201