[Abstract Title]. - Society for Neuroscience

Program#/Poster#: 236.15/D15
Topic: B.06.a. Docking and fusion
Support: NSF
Title: In vivo functions and interactions of NSF and SNAP in neurotransmitter release
Authors: W. YU 1 , F. KAWASAKI 2 , *R. W. ORDWAY 2 ;
1 Intercollege Grad. Program in Genet., 2 Biol & Ctr. Mol Cell Neurosci, Penn State Univ.,
University Park, PA
Abstract: Previous work has defined the in vivo role of the Drosophila N-ethylmaleimide-
Sensitive Factor 1 (dNSF1) protein in neurotransmitter release through analysis of a temperaturesensitive
(TS) paralytic dNSF1 mutant, comatose. One extension of these studies involves
analysis of dNSF1 tagged with enhanced green fluorescent protein (EGFP). Transgenic
expression of dNSF1-EGFP has allowed live fluorescence imaging of wild-type and TS mutant
dNSF1 at native neuromuscular synapses. Wild-type dNSF1-EGFP was found to rescue
comatose TS paralytic and lethal mutants and is enriched within presynaptic boutons as observed
for native dNSF1, indicating dNSF1-EGFP retains the functional properties of dNSF1. Because
NSF interacts with the Soluble NSF Attachment Proteins (SNAPs) to disassemble SNARE
protein complexes, which are thought to participate in synaptic vesicle fusion, complementary
studies involve imaging a fluorescent Drosophila SNAP protein (tdTomato-dSNAP). Like
dNSF1-EGFP, tdTomato-SNAP is enriched within presynaptic boutons of native synapses.
Ongoing live imaging studies are employing fluorescence resonance energy transfer (FRET) and
fluorescence recovery after photobleaching (FRAP) to define the distribution, mobility and
interactions of NSF and SNAP at native synapses. These properties may be examined at resting
synapses, during synaptic activity, and in the comatose (NSF) TS paralytic mutant to further
define the in vivo roles of NSF and SNAP in synaptic vesicle trafficking.
Disclosures: W. Yu, None; R.W. Ordway , None; F. Kawasaki, None.
Poster
236. Neurotransmitter Release: Docking and Fusion
Time: Sunday, November 16, 2008, 1:00 pm - 5:00 pm
Program#/Poster#: 236.16/D16
Topic: B.06.a. Docking and fusion
Support: Max-Planck Society