EXCEED THE SPACE PROVIDED. The objective of the proposed research is to understand the mechanisms by which neuronal activity regulates gene; expression. To achieve this goal we have characterized the signal transduction pathways by which Ca^+ influx through L-type voltage sensitive Ca^+ channels stimulates the transcription of two genes in neurons, the o-fos proto-oncogene and the gene encoding brain derived neurotrophic factor (BDNF). BDNF gene transcription is activated by Ca^+ influx into neurons, and like c-fos transcription BDNF transcription is controlled by the transcription factor CREB. The 40 kB BDNF gene gives rise to eight distinct mRNAs whose synthesis is driven by four separate promoters. Our studies during the previous funding period revealed that BDNF promoter III is the most Ca~+ responsive of the BDNF promoters.. A CREB binding site locaied at -35 nucleotides relative to the site of initiation of BDNF promoter III transcription is critical for Ca2+ induction of BDNF transcription. In addition, a second -60 CaRE within the BDNF promoter cooperates with the CREB binding site to confer a Ca^+ response. The -60 CaRE binds an apparently novel transcription factor that may confer neuronal specificity to the BDNF Ca^+ response. Despite progress towards defining the mechanisms by which Ca^ regulates c-fos and BDNF transcription, a great deal still remains to be learned. There are as yet uncharacterized phosphorylation events that appear to be critical for Ca2+ induction of CREB-dependent gene transcription, and several not yet well defined transcription factors that in addition to CREB are capable of conferring a Ca~+ response in neurons. These transcription factors include members of the MEF2 family, and the transcription factor that binds to the -60 CaRE of the BDNF gene-. We propose three specific aims to address these unresolved issues: 1) To characterize phosphorylation modifications of the CREB/CBP complex, that in addition to CREB Ser-133 phosphorylation play a critical role in mediating Ca^+ responses, 2) To purify, clone, and characterize the transcription factor(s) that binds the -60 CaRE within BDNF promoter III, 3) To characterize the mechanism(s) by which Ca2+ regulates MEF2 activity, and to establish the importance of MEF2 proteins as mediators of calcium's effects on gene expression. The proposed research will provide insight into how Ca^+ regulates gene transcription, and promotes critical cellularresponses that underlieneuronal differentiation, survival, and plasticity. PERFOFIMANCE SITE(S) (organization, dry, state) Children's Hospital Boston, Massachusetts KEY PERSONNEL ========================================Section End===========================================

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37NS028829-17
Application #
6921883
Study Section
Special Emphasis Panel (NSS)
Program Officer
Leblanc, Gabrielle G
Project Start
1990-08-01
Project End
2007-02-28
Budget Start
2005-07-01
Budget End
2007-02-28
Support Year
17
Fiscal Year
2005
Total Cost
$544,539
Indirect Cost
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Kalish, Brian T; Cheadle, Lucas; Hrvatin, Sinisa et al. (2018) Single-cell transcriptomics of the developing lateral geniculate nucleus reveals insights into circuit assembly and refinement. Proc Natl Acad Sci U S A 115:E1051-E1060
Van Schil, Kristof; Naessens, Sarah; Van de Sompele, Stijn et al. (2018) Mapping the genomic landscape of inherited retinal disease genes prioritizes genes prone to coding and noncoding copy-number variations. Genet Med 20:202-213
Cheadle, Lucas; Tzeng, Christopher P; Kalish, Brian T et al. (2018) Visual Experience-Dependent Expression of Fn14 Is Required for Retinogeniculate Refinement. Neuron 99:525-539.e10
Hrvatin, Sinisa; Hochbaum, Daniel R; Nagy, M Aurel et al. (2018) Single-cell analysis of experience-dependent transcriptomic states in the mouse visual cortex. Nat Neurosci 21:120-129
Yap, Ee-Lynn; Greenberg, Michael E (2018) Activity-Regulated Transcription: Bridging the Gap between Neural Activity and Behavior. Neuron 100:330-348
Vierbuchen, Thomas; Ling, Emi; Cowley, Christopher J et al. (2017) AP-1 Transcription Factors and the BAF Complex Mediate Signal-Dependent Enhancer Selection. Mol Cell 68:1067-1082.e12
Mardinly, A R; Spiegel, I; Patrizi, A et al. (2016) Sensory experience regulates cortical inhibition by inducing IGF1 in VIP neurons. Nature 531:371-5
Andzelm, Milena M; Cherry, Timothy J; Harmin, David A et al. (2015) MEF2D drives photoreceptor development through a genome-wide competition for tissue-specific enhancers. Neuron 86:247-63
Spiegel, Ivo; Mardinly, Alan R; Gabel, Harrison W et al. (2014) Npas4 regulates excitatory-inhibitory balance within neural circuits through cell-type-specific gene programs. Cell 157:1216-29
Malik, Athar N; Vierbuchen, Thomas; Hemberg, Martin et al. (2014) Genome-wide identification and characterization of functional neuronal activity-dependent enhancers. Nat Neurosci 17:1330-9

Showing the most recent 10 out of 45 publications