There is emerging evidence from in vitro systems that a remarkable number of mRNAs contain alternative translation initiation sites, further amplifying the number and diversity of protein products that can be generated from a single RNA molecule. This new diversity can be studied in a high throughput and systematic manner in cell culture using newly developed methods for """"""""ribosome footprinting."""""""" Here, we propose to bring these new methods into the study of the nervous system using our tools and expertise in capturing ribosomes from genetically defined cell populations in the mouse brain. Thus, Aim 1 is to determine if alternative translation is a common feature of the complex mammalian brain, and how it is regulated across distinct cell types. Furthermore, in yeast and other in vitro systems, alternative translation is regulated by cellular stress and other manipulations.
Our second aim i s to determine whether neuronal activity can regulate alternative translation in vivo, using a newly developed dual-reporter """"""""CHOP-TRAP"""""""" mouse that permits parallel optogenetic manipulation and ribosome capture from targeted cell populations, in this case midbrain Dopaminergic neurons. It is our hope that the development of tools to characterize alternative translation in cel populations relevant to reward and addiction will permit future in-depth analysis of this process in CNS disorder.

Public Health Relevance

It has recently been discovered in stem cells that a process called alternative translation, the making of multiple proteins from the same mRNA, occurs surprisingly often. The proposed project tests whether this same process is widespread in the brain, and whether activity in the circuits that regulate addiction and reward can alter this process.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21DA038458-01
Application #
8787911
Study Section
Special Emphasis Panel (ZDA1)
Program Officer
Satterlee, John S
Project Start
2014-07-01
Project End
2016-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Washington University
Department
Genetics
Type
Schools of Medicine
DUNS #
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
O'Connor, Shawn David; Cabrera, Omar HoseĆ”; Dougherty, Joseph D et al. (2017) Dexmedetomidine protects against glucocorticoid induced progenitor cell apoptosis in neonatal mouse cerebellum. J Matern Fetal Neonatal Med 30:2156-2162
Dougherty, Joseph D (2017) Generation and characterization of a mouse line for monitoring translation in dopaminergic neurons. Sci Rep 7:8117
Dougherty, Joseph D; Yang, Chengran; Lake, Allison M (2017) Systems biology in the central nervous system: a brief perspective on essential recent advancements. Curr Opin Syst Biol 3:67-76
Reddy, Adarsh S; O'Brien, David; Pisat, Nilambari et al. (2017) A Comprehensive Analysis of Cell Type-Specific Nuclear RNA From Neurons and Glia of the Brain. Biol Psychiatry 81:252-264
Ouwenga, Rebecca; Lake, Allison M; O'Brien, David et al. (2017) Transcriptomic Analysis of Ribosome-Bound mRNA in Cortical Neurites In Vivo. J Neurosci 37:8688-8705
Dougherty, Joseph D (2017) The Expanding Toolkit of Translating Ribosome Affinity Purification. J Neurosci 37:12079-12087
Sakers, Kristina; Lake, Allison M; Khazanchi, Rohan et al. (2017) Astrocytes locally translate transcripts in their peripheral processes. Proc Natl Acad Sci U S A 114:E3830-E3838
Dalal, Jasbir S; Yang, Chengran; Sapkota, Darshan et al. (2017) Quantitative Nucleotide Level Analysis of Regulation of Translation in Response to Depolarization of Cultured Neural Cells. Front Mol Neurosci 10:9
Maloney, Susan E; Khangura, Eakta; Dougherty, Joseph D (2016) The RNA-binding protein Celf6 is highly expressed in diencephalic nuclei and neuromodulatory cell populations of the mouse brain. Brain Struct Funct 221:1809-31
Kopp, Nathan; Climer, Sharlee; Dougherty, Joseph D (2015) Moving from capstones toward cornerstones: successes and challenges in applying systems biology to identify mechanisms of autism spectrum disorders. Front Genet 6:301

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