Over 95% of the neurons that make up the striatum (caudate and nucleus accumbens) are medium size spiny neurons (MSNs). All MSNs are dopaminoceptive; i.e. they receive dopamine input and express one or more of the dopamine receptor subtypes. Dysregulation of gene expression in MSNs is implicated in the pathophysiology and treatment of many neuropsychiatric diseases, including Huntington's disease, Parkinson's disease, drug addiction, affective disorders, attention deficit hyperactivity disorder, and schizophrenia; despite this, few details are available regarding how the differentiated MSN phenotype is specified at the molecular level. The dopamine and cyclic AMP-regulated phosphoprotein, 32 kDa (DARPP- 32) is the most widely used marker of the differentiated MSN, and DARPP-32 is a key modulator of MSN functions, especially those mediated by dopamine. DARPP-32 levels are highly influenced in vivo and in vitro by levels of brain-derived neurotrophic factor (BDNF'), and BDNF regulates DARPP-32 expression via the phosphatidylinositoi 3-kinase (PI3K) and p35/cdk5 pathways. The objectives of this proposal are:
SPECIFIC AIM 1 To identify the cis-acting sequence(s) within the 9Kb DARPP-32 genomic fragment identified in this laboratory that direct(s) transgene expression to the MSNs, using transgenesis, transient transfection, electrophoretic mobility shift assays and DNase footprinting, with particular attention to distinctions between striatal patch and matrix compartments.
SPECIFIC AIM 2 To determine in vitro: A) whether the DNA cis-acting regions mediating MSN-specific transcription also mediate regulation by BDNF and PI3-K within these neurons; B) the relationship between PI3-K, Akt and cdk5 in the regulation of DARPP-32 expression utilizing primary neuronal cultures, transfection, antisense oligonucleotides and viral transduction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS045942-02
Application #
6999861
Study Section
Special Emphasis Panel (ZRG1-MDCN-A (03))
Program Officer
Mamounas, Laura
Project Start
2005-01-01
Project End
2008-12-31
Budget Start
2006-01-01
Budget End
2006-12-31
Support Year
2
Fiscal Year
2006
Total Cost
$317,045
Indirect Cost
Name
Thomas Jefferson University
Department
Neurology
Type
Schools of Medicine
DUNS #
053284659
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Chandwani, Samira; Keilani, Serene; Ortiz-Virumbrales, Maitane et al. (2013) Induction of DARPP-32 by brain-derived neurotrophic factor in striatal neurons in vitro is modified by histone deacetylase inhibitors and Nab2. PLoS One 8:e76842
Keilani, Serene; Chandwani, Samira; Dolios, Georgia et al. (2012) Egr-1 induces DARPP-32 expression in striatal medium spiny neurons via a conserved intragenic element. J Neurosci 32:6808-18
Hager, Stefanie; Lösch, Saskia; Noll, Stephan et al. (2012) Red/ET recombination with chimeric oligonucleotides allows rapid generation of BAC transgenes harboring full-length or truncated huntingtin cDNA. Biotechniques 53:
Thomas, Elizabeth A; Coppola, Giovanni; Tang, Bin et al. (2011) In vivo cell-autonomous transcriptional abnormalities revealed in mice expressing mutant huntingtin in striatal but not cortical neurons. Hum Mol Genet 20:1049-60
Pedrini, Steve; Bogush, Alexey; Ehrlich, Michelle E (2008) Phosphatidylinositide 3-kinase and protein kinase C zeta mediate retinoic acid induction of DARPP-32 in medium size spiny neurons in vitro. J Neurochem 106:917-24
Bogush, Alexey; Pedrini, Steve; Pelta-Heller, Joshua et al. (2007) AKT and CDK5/p35 mediate brain-derived neurotrophic factor induction of DARPP-32 in medium size spiny neurons in vitro. J Biol Chem 282:7352-9
Bogush, Alexey I; McCarthy, Lois E; Tian, Chai et al. (2005) DARPP-32 genomic fragments drive Cre expression in postnatal striatum. Genesis 42:37-46