The main focus of this research proposal is to determine the molecular mechanisms by which spontaneous neurotransmitter release is regulated. Characterization of neuronal communication at synapses has largely focused on action potential-triggered synaptic vesicle fusion, with spontaneous miniature potentials (minis) largely thought to represent background noise. However, spontaneous release is regulated independently of evoked release. Moreover, the frequency of spontaneous release is regulated by activity and can drive synaptic structural modification and growth, changes resulting in long-lasting alterations in neuronal connectivity. Regulation of spontaneous release would likely impinge directly on the molecular """"""""fusion clamp"""""""" which prevents primed vesicles from fusing with the presynaptic membrane in the absence of calcium, Complexin has been identified as the vesicle fusion clamp that regulates spontaneous release at the Drosophila neuromuscular junction (NMJ). However, the molecular mechanism by which complexin functions as a fusion clamp to regulate spontaneous release is unknown. The Drosophila NMJ will be used as an in vivo model system to determine the molecular mechanisms by which complexin regulates spontaneous neurotransmitter release.
Aim 1 will examine evolutionary conservation of complexin function as a vesicle fusion clamp.
Aim 2 will define the molecular mechanisms that mediate complexin's ability to regulate spontaneous release.
Both aims will make extensive use of genetic tools available in Drosophila and in vivo electrophysiology recordings, as well as in vitro biochemical and immunocytochemical approaches. Alterations in complexin levels have been reported in a number of neurological diseases including schizophrenia, Huntington's disease, and Alzheimer's disease, suggesting the complexin dysfunction and abnormal rates of spontaneous release may contribute to several human neuropathologies. Defects in neurotransmitter release are implicated in a number of neurological diseases including schizophrenia, Huntington's disease, and Alzheimer's disease. Understanding the molecular mechanisms that underlie neurotransmitter release will provide potentially new targets and insights into how altered spontaneous release contributes to neurological diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32NS064750-02
Application #
8016060
Study Section
Special Emphasis Panel (ZRG1-F03B-H (20))
Program Officer
Talley, Edmund M
Project Start
2009-08-05
Project End
2012-08-04
Budget Start
2010-08-05
Budget End
2011-08-04
Support Year
2
Fiscal Year
2010
Total Cost
$53,810
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Miscellaneous
Type
Schools of Arts and Sciences
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Cho, Richard W; Buhl, Lauren K; Volfson, Dina et al. (2015) Phosphorylation of Complexin by PKA Regulates Activity-Dependent Spontaneous Neurotransmitter Release and Structural Synaptic Plasticity. Neuron 88:749-61
Cho, Richard W; K├╝mmel, Daniel; Li, Feng et al. (2014) Genetic analysis of the Complexin trans-clamping model for cross-linking SNARE complexes in vivo. Proc Natl Acad Sci U S A 111:10317-22
Cho, Richard W; Song, Yun; Littleton, J Troy (2010) Comparative analysis of Drosophila and mammalian complexins as fusion clamps and facilitators of neurotransmitter release. Mol Cell Neurosci 45:389-97