Neural circuits are organized to elicit appropriate behavioral responses to environmental stimuli. Advancement in technologies allowing the visualization and manipulation of neural activity has made it possible to establish a causal relationship between circuit function and behavior. This proposal attempts to expand the repertoire of existing technologies by exploiting a unique feature of the NFAT molecule - calcium/calcineurin-dependent nuclear localization. In the novel activity reporter system named CaLexA, a truncated version of NFAT containing the necessary nuclear localization signal is fused to LexA to make the synthetic transcription factor LexA- NFAT. The resulting activity reporter system will detect not only persistent neural activity but phasic activity as well with hih sensitivity in freely moving, behaving animals. We have identified several state-of-the-art molecular technologies that will increase the signal- to-noise ratio of the CaLexA activity reporter system as well as add the ability to discriminate between phasic and sustained neural activity.
In Aim 1, we will engineer new LexA-NFAT constructs to suppress unwanted background and enhance behaviorally relevant signal.
In Aim 2, we will design new LexA-NFAT constructs that will allow the detection of transient calcium activity.
In Aim 3, we will use optogenetics to generate precisely controlled patterns of neural activity so that calcium imaging and the CaLexA reporter system can be compared for their sensitivity and dynamic range to gain a full characterization of all the LexA-NFAT variants. The creation and characterization of these new CaLexA activity reporter systems will make available to the Drosophila neurobiology community a new set of tools for the study of neural circuits and behavior, and molecular designs that can be readily modified for other genetic model organisms.

Public Health Relevance

How the action of neurons, synapses, and circuits underlies brain function and dysfunction is a fundamental question for neuroscience. The objective of the proposed study is to develop a novel genetic system to mark, map and manipulate active neurons in behaving animals. The methods described in this application will make available to the Drosophila neurobiology community a new set of tools for the study of neural circuits and behavior, and molecular designs that can be readily modified for other genetic model organisms. The work of this proposal is basic science that seeks to reveal the underlying circuit principle for cognitive behavior and sensory perception, creating a knowledge base from which new cellular targets can be evaluated for future therapeutic interventions of psychiatric disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21MH106958-02
Application #
9022518
Study Section
Molecular Neurogenetics Study Section (MNG)
Program Officer
Asanuma, Chiiko
Project Start
2015-02-25
Project End
2017-01-31
Budget Start
2016-02-01
Budget End
2017-01-31
Support Year
2
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093