? P002 Several genome-wide studies, including studies from the previous funding period of the PARC, identified the extracellular matrix (ECM) and astrocytes as being involved in both the risk for and the effects of ethanol drinking. Cells in the brain parenchyma are separated by the extracellular space, accounting for approximately 20% of the total volume of the mature rodent brain and containing a highly organized ECM that forms an insoluble network around cells. The brain interstitial ECM significantly contributes to the molecular signals regulating neuronal plasticity. Recent research has shown that astrocytes are major producers of the brain interstitial ECM as well as ECM proteases, which are involved in the remodeling of the ECM. ECM proteins are post-translationally modified by glycosylation. Proteoglycans are major components of the brain ECM and consist of a core protein covalently bound to glycosaminoglycans (GAGs) formed by repeating disaccharides, modified by sulfation. Hyaluronic acid (HA), also a major component of the ECM, is a non-sulfated GAG and is not covalently bound to proteins. GAGs are involved in the modulation of neuronal plasticity. The overall goal of this proposal is to identify differential expression of astrocyte-specific genes (with a major emphasis on genes involved in the formation and remodeling of the ECM) and differential levels of ECM GAGs associated with ethanol drinking in two brain regions involved in addiction: the medial prefrontal cortex (mPFC; including the prelimbic and infralimbic cortex) and the nucleus accumbens (NAc). We plan to reach this overarching goal by pursuing the following two specific aims: 1) Definition of the astrocyte translatome in the mPFC and NAc of nave and ethanol-drinking Aldh1l1-EGFP-Rpl10a mice; validation of differential translation and expression by qPCR and by RNA scope, respectively, in Aldh1l1-EGFP-Rpl10a mice; determination of the expression of selected ECM-related genes in ethanol nave high ethanol preference (HP) and low ethanol preference (LP) mice, and in ethanol-drinking HP mice by RNAscope. We will use the translating ribosome affinity purification (TRAP) technology in Aldh1l1-EGFP-Rpl10a mice that express a modified ribosomal protein Rpl10a with an eGFP tag (EGFP-Rpl10a) only in cells expressing the astrocytic marker Aldh1l1. Selected differentially translated RNAs for ECM-related proteins will be confirmed by qPCR and their differential expression and sub- regional localization will be determined by RNAscope. Differential expression of selected ECM-related genes will also be determined in ethanol nave HP vs. LP mice to investigate genes involved in the risk of ethanol drinking and in drinking and nave HP mice to investigate the effects of ethanol drinking in this selected line by RNAscope. 2) Determination of chondroitin sulfate (CS)-, heparan sulfate (HS)-, and HA-GAG disaccharide levels in the mPFC and NAc of ethanol nave and ethanol-drinking Aldh1l1-EGFP-Rpl10a and HP mice and in nave LP mice. The proposed studies will characterize the role of astrocyte ECM and of sugar chains in the risk and consequences of ethanol drinking and identify cellular and molecular targets for intervention.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Comprehensive Center (P60)
Project #
2P60AA010760-26
Application #
10056067
Study Section
Special Emphasis Panel (ZAA1)
Project Start
1996-12-01
Project End
2025-12-31
Budget Start
2021-01-01
Budget End
2021-12-31
Support Year
26
Fiscal Year
2021
Total Cost
Indirect Cost
Name
Oregon Health and Science University
Department
Type
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Morales, Angelica M; Jones, Scott A; Ehlers, Alissa et al. (2018) Ventral striatal response during decision making involving risk and reward is associated with future binge drinking in adolescents. Neuropsychopharmacology 43:1884-1890
Gavin, David P; Hashimoto, Joel G; Lazar, Nathan H et al. (2018) Stable Histone Methylation Changes at Proteoglycan Network Genes Following Ethanol Exposure. Front Genet 9:346
Purohit, Kush; Parekh, Puja K; Kern, Joseph et al. (2018) Pharmacogenetic Manipulation of the Nucleus Accumbens Alters Binge-Like Alcohol Drinking in Mice. Alcohol Clin Exp Res 42:879-888
Müller-Oehring, Eva M; Kwon, Dongjin; Nagel, Bonnie J et al. (2018) Influences of Age, Sex, and Moderate Alcohol Drinking on the Intrinsic Functional Architecture of Adolescent Brains. Cereb Cortex 28:1049-1063
Iancu, Ovidiu Dan; Colville, Alex M; Wilmot, Beth et al. (2018) Gender-Specific Effects of Selection for Drinking in the Dark on the Network Roles of Coding and Noncoding RNAs. Alcohol Clin Exp Res :
Kafkafi, Neri; Agassi, Joseph; Chesler, Elissa J et al. (2018) Reproducibility and replicability of rodent phenotyping in preclinical studies. Neurosci Biobehav Rev 87:218-232
Qiu, J; Wagner, E J; Rønnekleiv, O K et al. (2018) Insulin and leptin excite anorexigenic pro-opiomelanocortin neurones via activation of TRPC5 channels. J Neuroendocrinol 30:
Allen, Daicia C; Ford, Matthew M; Grant, Kathleen A (2018) Cross-Species Translational Findings in the Discriminative Stimulus Effects of Ethanol. Curr Top Behav Neurosci 39:95-111
Aoun, E G; Jimenez, V A; Vendruscolo, L F et al. (2018) A relationship between the aldosterone-mineralocorticoid receptor pathway and alcohol drinking: preliminary translational findings across rats, monkeys and humans. Mol Psychiatry 23:1466-1473
Colville, Alexandre M; Iancu, Ovidiu D; Lockwood, Denesa R et al. (2018) Regional Differences and Similarities in the Brain Transcriptome for Mice Selected for Ethanol Preference From HS-CC Founders. Front Genet 9:300

Showing the most recent 10 out of 291 publications