The GABA intemeuron plays a central role in integrating cortical output and is emerging as a major locus of schizophrenia molecular pathology. Reelin (RELN), GAD67 and DNA methylating enzyme (DNMT1) are all expressed in GABA interneurons, and are each abnormally regulated in schizophrenic brain. Further, DNMT1 is a critical component of the epigenetic regulatory cascade, now shown to play an essential role in neuronal gene expression. We have earlier reported on the effects of epigenetic modifiers (DNA methylation, chromatin structure, and methylated-DNA binding proteins) on the levels of RELN and GAD67 expression. This career development award will extend these observations using primary neuronal cultures (PNC) to describe variations in epigenetic modifications of the RELN and GAD67 promoters resulting from pharmacological treatment and membrane depolarization. Supporting the use of PNCs, we and others have observed El4 mouse neuron cultures to be predominantly GABAergic in phenotype (co-expressing RELN, GAD and DNMT1). Our first objective, using these homogenous neuronal cultures, will be to analyze DNA methylation, local chromatin structure and promoter DNA-protein interactions in response to pharmacological treatments with known epigenetic effects, specifically L-methionine, a hypermethylating stimulus, and histone deactylase (HDAC) inhibitors such as valproic acid. Supporting this first objective is our earlier demonstration that L-methionine down regulates RELN and GAD67 mRNA expression as a consequence of epigenetic modifications along their promoters, and this is reversed by the application of valproic acid. Our second objective will be to examine the effects of membrane depolarization on epigenetic modification of depolarization inducible promoters within these GABAergic neurons. We will apply antisense oligonucleotide strategies to down regulate epigenetic repressor proteins such as DNMT1 and MeCP2 to dissect their contribution to depolarization induced gene transcription. Supporting this second objective, we have now shown that antisense induced down regulation of DNMT1 mRNA and protein will result in significant increases in RELN mRNA expression. Also, we have preliminary data indicating robust changes in GAD67 and DNMT1 mRNA in response to depolarization. With this KO1 career development award, the candidate seeks training in focused methods in the molecular study of epigenetic gene regulation. Because epigenetic mechanisms are modifiable by drug interventions, this work has immediate relevance to the development of novel therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01MH069839-05
Application #
7619304
Study Section
Neural Basis of Psychopathology, Addictions and Sleep Disorders Study Section (NPAS)
Program Officer
Desmond, Nancy L
Project Start
2005-04-01
Project End
2011-03-31
Budget Start
2009-04-01
Budget End
2011-03-31
Support Year
5
Fiscal Year
2009
Total Cost
$177,179
Indirect Cost
Name
University of Illinois at Chicago
Department
Psychiatry
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Chase, Kayla; Sharma, Rajiv P (2013) Epigenetic developmental programs and adipogenesis: implications for psychotropic induced obesity. Epigenetics 8:1133-40
Sharma, R P; Gavin, D P; Chase, K A (2012) Heterochromatin as an incubator for pathology and treatment non-response: implication for neuropsychiatric illness. Pharmacogenomics J 12:361-7
Gavin, David P; Sharma, Rajiv P; Chase, Kayla A et al. (2012) Growth arrest and DNA-damage-inducible, beta (GADD45b)-mediated DNA demethylation in major psychosis. Neuropsychopharmacology 37:531-42
Gavin, David P; Chase, Kayla A; Sharma, Rajiv P (2011) Enhancement of psychotherapy using epigenetic modulating drugs. Med Hypotheses 77:121-4
Sharma, Rajiv P; Gavin, David P; Grayson, Dennis R (2010) CpG methylation in neurons: message, memory, or mask? Neuropsychopharmacology 35:2009-20
Gavin, David P; Sharma, Rajiv P (2010) Histone modifications, DNA methylation, and schizophrenia. Neurosci Biobehav Rev 34:882-8
Imbesi, Marta; Arslan, Ahmet Dirim; Yildiz, Sevim et al. (2009) The melatonin receptor MT1 is required for the differential regulatory actions of melatonin on neuronal 'clock'gene expression in striatal neurons in vitro. J Pineal Res 46:87-94
Imbesi, M; Yildiz, S; Dirim Arslan, A et al. (2009) Dopamine receptor-mediated regulation of neuronal ""clock"" gene expression. Neuroscience 158:537-44
Sharma, Rajiv P; Grayson, Dennis R; Gavin, David P (2008) Histone deactylase 1 expression is increased in the prefrontal cortex of schizophrenia subjects: analysis of the National Brain Databank microarray collection. Schizophr Res 98:111-7
Imbesi, Marta; Uz, Tolga; Manev, Radmila et al. (2008) Minocycline increases phosphorylation and membrane insertion of neuronal GluR1 receptors. Neurosci Lett 447:134-7

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