Here we propose to develop an experimental paradigm to allow dynamic monitoring of the strength and location of every glutamatergic and GABA/Glycinergic synapse within the brain of a living organism. In combination with behavioral manipulation of the organism this paradigm will allow for study of how the brain encodes information in synaptic structure. This paradigm will involve combining three technologies: 1. Recombinant probes with which postsynaptic excitatory and inhibitory sites can be labeled in vivo, allowing the location and strength of synaptic connections to be monitored in parallel. 2. 2P-SPIM microscopy, which can image large volumes very quickly, without bleaching and, potentially, with isotropic resolution. 3. Software to calculate and store the location and strengt of each synapse in such a manner that it can be easily manipulated and analyzed. Experiments will be performed in zebrafish, as they have semi-transparent brains that are relatively small, yet they are capable of relatively complex behaviors. Experiments will be used both to establish the viability of the paradigm and to answer fundamental questions about how synapses are modulated during sleep, as well as how they are changed in learning paradigms such as sound habituation and place preference/aversion conditioning.

Public Health Relevance

The purpose of the experiments described in the grant is to develop technology capable of mapping the location, strength and polarity of all the excitatory and inhibitory synapses in the brain of a living animal at different points in time. We will use recombinant probes that can mark synapses in vivo, 2p-SPIM microscopy to image the probes and big-data analytics-based software for analyzing data.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
1R01MH107238-01
Application #
8754412
Study Section
Special Emphasis Panel (ZRG1-BCMB-A (51))
Program Officer
Freund, Michelle
Project Start
2014-09-25
Project End
2019-07-31
Budget Start
2014-09-25
Budget End
2015-07-31
Support Year
1
Fiscal Year
2014
Total Cost
$1,885,883
Indirect Cost
$465,092
Name
University of Southern California
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
072933393
City
Los Angeles
State
CA
Country
United States
Zip Code
90089