Table of Contents
Mechanisms of Synaptic Vesicle Exocytosis
Early Composite drawing of synapse from EM
The Vesicle Cycle
Synaptic Transmission
Exocytosis is the "main" event
Special Needs of Synapses Satisfied
Modern Investigation of the SV Cycle
Small Synaptic Vesicle Cycle
Synaptic Vesicle Membrane
and Protein Content
Maturation of a SV
Isolation of Synaptic Vesicles by cell fractionation and centrifugation
Proteins of Synaptic Vesicles are
Isolated from Purified Synaptosomes
Step 1:
Vesicle filling with NT
EM immunogold to label NX in vesicles
Families of putative trafficking proteins
Step 2: Translocation to PM
Synapsins
Translocation?
3 steps of Exocytosis
Docking, Priming/Prefusion, Fusion/Exocytosis
Step 3: Synaptic Vesicle Docking
Step 3: Synaptic Vesicle Docking
Step 3: Candidates for Positioning SV at AZ
Synapsins
Attachment at AZ?
Mechanisms of Synaptic Vesicle Exocytosis cont.
Small Synaptic Vesicle Cycle
Model - Role of SNAREs in Exocytosis
Step 4: Priming/Prefusion Zipper Model
Structural domains of
SNARE Core Complex Proteins
Munc18 "holds" Syntaxin in a closed conformation, preventing core-complex formation
Step 5: Fusion/exocytosis
Calcium Influx During Transmission at the Squid Giant Synapse
How does Calcium Trigger NX Release?
Ca-binding proteins involved in exocytosis
Synaptotagmin - best candidate calcium sensor
Reliable - Unreliability of NX Release
Rab3 regulation of exocytosis
Evidence that Rab3 regulates exocytosis
Recovery of SNARE Core Complex Proteins
Synaptic Transmission
Synthetic Pathway for LDCV
Colocalization of SVs and LDCVs
Glutamate Transport - Anion
Transport of dopamine
(similar for other CA) into LDCVs
DA - NA in Chromaffin Granules
Locations of LDCV in CNS
Locations of LDCV
Delay to exocytosis
SVs release at low Hz, LDCVs at High HZ Stimulation
Release of LDCV
LDCV from Tom Sudhof email
Chromaffin (PC12 cells) Model System
Small Synaptic Vesicle Cycle
Evidence for partial release
Evanescent Near Field Microscopy
Evanescent Near Field Microscopy
Zenisek, Steyer, Almers Nature 2000 406:849
Do this whole paper and also some of the recent ones, maybe after Endocytosis
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