Molecular fingerprinting of VNUT-containing compartments in Neuro-2a cells

Author: 
Date created: 
2021-09-17
Identifier: 
etd21428
Keywords: 
SLC17A9
Purinergic signalling
Sympathetic co-transmission
VNUT-EGFP
Immunocytochemistry
Nearest-neighbour colocalization analysis
Abstract: 

Vesicular ATP release is involved in regulating biological processes like nociception, blood glucose, and vascular tone. ATP-containing vesicles are filled by the vesicular nucleotide transporter (VNUT) and are molecularly distinct from catecholaminergic vesicles of sympathetic neurons. This work sought to identify the molecular fingerprint of VNUT-containing vesicles. Fluorescence microscopy in Neuro-2a, HeLa, and HEK293 cells showed VNUT being widely dispersed throughout the cells with a perinuclear enrichment. VNUT failed to colocalize with known markers of synaptic vesicles, lysosomes, dense cored vesicles, and catecholaminergic vesicles. Bioinformatic analyses of mammalian VNUT C-terminus identified a unique KDEL-like HEDL motif, as well as a lack of classic synaptic vesicle-targeting dileucine-like and tyrosine-based motifs. This work suggests that VNUT likely resides primarily in the Golgi-ER complex, a previously unconsidered location for what is thought to be a vesicle-associated translocase.

Document type: 
Thesis
Rights: 
This thesis may be printed or downloaded for non-commercial research and scholarly purposes. Copyright remains with the author.
File(s): 
Supervisor(s): 
Damon Poburko
Department: 
Science: Department of Biomedical Physiology and Kinesiology
Thesis type: 
(Thesis) M.Sc.
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