Glutamate is the major excitatory neurotransmitter in the brain. The development and function of glutamatergic synapses is essential for proper neuronal connectivity and brain function, including learning and memory. Recent data suggest that defects in synapse development and function contribute to several neurodevelopmental and neuropsychiatric diseases such as intellectual disability, autism spectrum disorders, Alzheimer's disease and schizophrenia. In addition, excessive activation of glutamate signaling can lead to excitotoxic cell death in ischemia, stroke and neurodegenerative diseases. Thus, it is important to understand the cell biological and molecular mechanisms involved in regulating glutamatergic synapses. While much progress has been made identifying genes that regulate glutamatergic synapse development and function in vitro using neuronal cultures, less is known about the in vivo function of many of those genes. Challenges of in vivo analysis include the fact that mouse gene knock-outs do not always lead to the same dramatic synaptic defects observed in vitro, perhaps due to gene redundancy, and furthermore, compound knock-outs of gene families often lead to embryonic lethality further hindering phenotypic analyses. We use C. elegans as a genetic model to study genes and mechanisms that regulate glutamatergic synapses in vivo. Advantages of C. elegans include less gene redundancy, powerful genetic tools, a simple defined nervous system, and the ability of the animal to tolerate severe reductions in neuronal function. The goal of this exploratory proposal is to identify novel genes and mechanisms that regulate glutamatergic synapse development and function in vivo.
In Aim 1, we combine several strengths of C. elegans and develop an innovative, optogenetic behavioral screen, based on a simple glutamatergic behavior, to identify conserved neuronal genes that regulate glutamatergic synapes. We have completed a pilot screen of genes with cell-adhesion molecule domains and identified several strong candidates, including the Ig-domain-containing, VEGF (Vascular Endothelial Growth Factor) receptor-related genes, ver-1 and ver-4.
In Aim 2, we investigate the role of ver-1 and ver-4 in regulating glutamatergic synapses to illustrate our strategy for analyzing top candidates from our screen. Understanding how VEGFR signaling regulates glutamatergic synapses in C. elegans will be informative given that worms do not possess a cardiovascular system where VEGF has traditionally been shown to act. Identifying novel genes and fundamental mechanisms that regulate glutamatergic synapses may provide potential therapeutic targets for treatment of neurological diseases.

Public Health Relevance

The development and function of glutamatergic synapses is essential for proper synaptic connectivity and brain function. Defects in synapse development and function contribute to several neurodevelopmental and neuropsychiatric diseases including autism spectrum disorders, Alzheimer's disease and schizophrenia, and excessive glutamatergic signaling contributes to excitotoxic cell death in ischemia and stroke. This research is focused on identifying novel genes and mechanisms required for glutamatergic synapse development and function, which may reveal new drug targets for the treatment of neurological diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21NS101534-02
Application #
9415103
Study Section
Synapses, Cytoskeleton and Trafficking Study Section (SYN)
Program Officer
Morris, Jill A
Project Start
2017-02-01
Project End
2019-01-31
Budget Start
2018-02-01
Budget End
2019-01-31
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Tufts University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02111
Hodul, Molly; Dahlberg, Caroline L; Juo, Peter (2017) Function of the Deubiquitinating Enzyme USP46 in the Nervous System and Its Regulation by WD40-Repeat Proteins. Front Synaptic Neurosci 9:16