The N-methyl-D-aspartate (NMDA) receptor, an excitatory neurotransmitter receptor in the brain, is an important site of action of ethanol. Following chronic ethanol treatment in vivo and in vitro, NMDA receptor number and function are upregulated, with a concomitant increase in R1 and R2B polypeptide levels in vitro. Similar ethanol treatment in vitro increases R1 mRNA half-life from 16 h to more than 24 h (Kumari and Ticku, 1998a) indicating that post-transcriptional mechanisms operate to augment NMDA receptor number in cortical neurons exposed to chronic ethanol treatment (50 mM, 5 days). More recently, we observed that de novo protein synthesis is required for ethanol-induced stabilization of R1 mRNA (Kumari and Ticku, 1998b), suggesting that ethanol-induced unknown protein factor(s) mediate this effect. Long term plans of this project are to elucidate the post-transcriptional mechanisms involved in the stabilization of NMDA R1 mRNA in fetal cortical neurons exposed to chronic ethanol treatment. Hypothesis to be tested in this proposal are (1) to identify specific RNA sequences (or cis-acting regulatory elements) of the R1 mRNA; and, (2) the nature of ethanol-induced cytoplasmic protein(s) (or trans-acting factors) that interact with cis- acting RNA regulatory sequences. These objectives will be achieved by (a) examining whether ethanol induces transcription of a stable R1 splice-variant; (b) delineating the cis -acting regulatory region(s) within the primary sequence of the R1 mRNA using cell-free mRNA decay assay and cell transfections; (c) dissecting the cis-acting sequences within the regulatory region defined above using mutants created by nested deletion, linker scanning and base substitution coupled to RNA gel shift assays and cell transfections, and finally (d) identifying the nature of trans-acting factor(s) by UV cross-linking and Northwestern analysis. A more thorough understanding of the pertinent molecular mechanisms through which ethanol modulates NMDA R1 mRNA stability may permit the design of novel therapeutic approaches to alcohol-related diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
5R01AA012070-03
Application #
6349712
Study Section
Special Emphasis Panel (ZRG4-ALTX-3 (01))
Project Start
1999-02-01
Project End
2003-01-31
Budget Start
2001-02-01
Budget End
2002-01-31
Support Year
3
Fiscal Year
2001
Total Cost
$145,841
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Pharmacology
Type
Other Domestic Higher Education
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Anji, Antje; Kumari, Meena (2016) Guardian of Genetic Messenger-RNA-Binding Proteins. Biomolecules 6:4
Anji, Antje; Miller, Hayley; Raman, Chandrasekar et al. (2015) Expression of ?-subunit of ?-glucosidase II in adult mouse brain regions and selected organs. J Neurosci Res 93:82-93
Velders, Fleur P; Kuningas, Maris; Kumari, Meena et al. (2011) Genetics of cortisol secretion and depressive symptoms: a candidate gene and genome wide association approach. Psychoneuroendocrinology 36:1053-61
Zabaneh, Delilah; Kumari, Meena; Sandhu, Manj et al. (2011) Meta analysis of candidate gene variants outside the LPA locus with Lp(a) plasma levels in 14,500 participants of six White European cohorts. Atherosclerosis 217:447-51
Anji, Antje; Kumari, Meena (2011) A cis-acting region in the N-methyl-d-aspartate R1 3'-untranslated region interacts with the novel RNA-binding proteins beta subunit of alpha glucosidase II and annexin A2--effect of chronic ethanol exposure in vivo. Eur J Neurosci 34:1200-11
Anji, Antje; Kumari, Meena (2009) Differentiated P19 cells express N-methyl-D-aspartate receptor 1 mRNA binding trans-acting proteins and four N-methyl-D-aspartate receptor 1 splice variants comparable to those in cultured fetal cortical neurons. J Neurosci Res 87:1591-601
Anji, Antje; Shaik, Kamran A; Kumari, Meena (2003) Effect of ethanol on lipid-mediated transfection of primary cortical neurons. Ann N Y Acad Sci 993:95-102; discussion 123-4
Kumari, Meena; Anji, Antje; Woods Jr, Henri et al. (2003) The molecular effects of alcohol: clues to the enigmatic action of alcohol. Ann N Y Acad Sci 993:82-94; discussion 123-4
Kumari, M (2001) Differential effects of chronic ethanol treatment on N-methyl-D-aspartate R1 splice variants in fetal cortical neurons. J Biol Chem 276:29764-71