Localization of the glycine transporter GLYT1 in glutamatergic synaptic vesicles

•A significant fraction of GLYT1 is located within neuronal terminals.•A purified synaptic vesicle fraction contains GLYT1.•Most of the GLYT1 containing vesicles also contain the glutamatergic marker vGLUT1. We have previously shown the presence of the glycine transporter GLYT1 in glutamatergic term...

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Bibliographic Details
Published in:Neurochemistry international 2014-07, Vol.73, p.204-210
Main Authors: Cubelos, Beatriz, Leite, Cristiana, Giménez, Cecilio, Zafra, Francisco
Format: Article
Language:English
Subjects:
Animals
Cell Membrane - metabolism
Cell Membrane - ultrastructure
Glutamate
Glutamic Acid - physiology
Glycine
Glycine Plasma Membrane Transport Proteins - isolation & purification
Glycine Plasma Membrane Transport Proteins - metabolism
GLYT1
Rats
Rats, Wistar
Synaptic vesicles
Synaptic Vesicles - metabolism
Synaptic Vesicles - ultrastructure
Transporter
Vesicular Glutamate Transport Protein 1 - metabolism
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Summary:•A significant fraction of GLYT1 is located within neuronal terminals.•A purified synaptic vesicle fraction contains GLYT1.•Most of the GLYT1 containing vesicles also contain the glutamatergic marker vGLUT1. We have previously shown the presence of the glycine transporter GLYT1 in glutamatergic terminals of the rat brain. In this study we present immunohistochemical and biochemical evidence indicating that GLYT1 is expressed not only at the plasma membrane of glutamatergic neurons, but also at synaptic vesicles. Confocal microscopy, immunoblots analysis of a highly purified synaptic vesicle fraction and immunoisolation of synaptic vesicles with anti-synaptophysin antibodies strongly suggested the presence of GLYT1 in synaptic vesicles. Moreover, direct observation with the electron microscope of purified vesicles immunoreacted with anti-GLYT1 and colloidal gold demonstrated that about 40% of the small vesicles of the purified vesicle fraction contained GLYT1. Double labeling for GLYT1 and synaptophysin of this vesicular fraction revealed that more of ninety percent of them were synaptic vesicles. Moreover, a significant part of the GLYT1 containing vesicles (86%) also contained the vesicular glutamate transporter vGLUT1, suggesting a functional role of GLYT1 in a subpopulation of glutamatergic vesicles.
ISSN:0197-0186
1872-9754
DOI:10.1016/j.neuint.2013.09.002