title: Changes in the Adult GluN2B Associated Proteome following Adolescent Intermittent Ethanol Exposure.
abstract: Adolescent alcohol use is the strongest predictor for alcohol use disorders. In rodents, adolescents have distinct responses to acute ethanol, and prolonged alcohol exposure during adolescence can maintain these phenotypes into adulthood. One brain region that is particularly sensitive to the effects of both acute and chronic ethanol exposure is the hippocampus. Adolescent intermittent ethanol exposure (AIE) produces long lasting changes in hippocampal synaptic plasticity and dendritic morphology, as well as in the susceptibility to acute ethanol-induced spatial memory impairment. Given the pattern of changes in hippocampal structure and function, one potential target for these effects is the ethanol sensitive GluN2B subunit of the NMDA receptor, which is known to be involved in synaptic plasticity and dendritic morphology. Thus we sought to determine if there were persistent changes in hippocampal GluN2B signaling cascades following AIE. We employed a previously validated GluN2B-targeted proteomic strategy that was used to identify novel signaling mechanisms altered by chronic ethanol exposure in the adult hippocampus. We collected adult hippocampal tissue (P70) from rats that had been given 2 weeks of AIE from P30-45. Tissue extracts were fractionated into synaptic and non-synaptic pools, immuno-precipitated for GluN2B, and then analyzed using proteomic methods. We detected a large number of proteins associated with GluN2B. AIE produced significant changes in the association of many proteins with GluN2B in both synaptic and non-synaptic fractions. Intriguingly the number of proteins changed in the non-synaptic fraction was double that found in the synaptic fraction. Some of these proteins include those involved in glutamate signaling cytoskeleton rearrangement, calcium signaling, and plasticity. Disruptions in these pathways may contribute to the persistent cellular and behavioral changes found in the adult hippocampus following AIE. Further, the robust change in non-synaptic proteins suggests that AIE may prime this signaling pathway for future ethanol exposures in adulthood.
authors.affiliation: Durham VA Medical Center, Duke University Medical Center, Durham, North Carolina, United States of America.|Department of Psychiatry and Behavioral Sciences, Duke University Medical Center, Durham, North Carolina, United States of America.|Department of Psychology and Neuroscience, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: H Scott
authors.initials: HS
authors.lastname: Swartzwelder
authors.affiliation: Durham VA Medical Center, Duke University Medical Center, Durham, North Carolina, United States of America.|Department of Psychiatry and Behavioral Sciences, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Mary-Louise
authors.initials: ML
authors.lastname: Risher
authors.affiliation: Durham VA Medical Center, Duke University Medical Center, Durham, North Carolina, United States of America.|Department of Psychiatry and Behavioral Sciences, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Kelsey M
authors.initials: KM
authors.lastname: Miller
authors.affiliation: Department of Molecular Physiology & Biophysics, Vanderbilt University, Nashville, TN, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, United States of America.|J. F. Kennedy Center for Research on Human Development, Vanderbilt University, Nashville, TN, United States of America.
authors.forename: Roger J
authors.initials: RJ
authors.lastname: Colbran
authors.affiliation: Department of Molecular Physiology & Biophysics, Vanderbilt University, Nashville, TN, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, United States of America.|J. F. Kennedy Center for Research on Human Development, Vanderbilt University, Nashville, TN, United States of America.
authors.forename: Danny G
authors.initials: DG
authors.lastname: Winder
authors.affiliation: Department of Molecular Physiology & Biophysics, Vanderbilt University, Nashville, TN, United States of America.|Department of Cell Biology and Anatomy, Louisiana State University Health Science Center, New Orleans, LA, United States of America.
authors.forename: Tiffany A
authors.initials: TA
authors.lastname: Wills
mesh_terms: D000293:Adolescent; D000818:Animals; D004195:Disease Models, Animal; D000431:Ethanol / Q000009:adverse effects*; D005260:Female; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008297:Male; D060066:Protein Interaction Maps / Q000187:drug effects*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D000066608:Underage Drinking
chemical_list: C121001:NR2B NMDA receptor; D020543:Proteome; D016194:Receptors, N-Methyl-D-Aspartate; D000431:Ethanol