Abstract
We expressed the synaptic vesicle proteins SV2, synaptotagmin, and synaptophysin in CHO fibroblasts to investigate the targeting information contained by each protein. All three proteins entered different cellular compartments. Synaptotagmin was found on the plasma membrane. Both SV2 and synaptophysin were sorted to small intracellular vesicles, but synaptophysin colocalized with early endosomal markers, while SV2 did not. SV2-containing vesicles did not have the same sedimentation characteristics as authentic synaptic vesicles, even though transfected SV2 was sorted from endosomal markers. We also created cell lines expressing both SV2 and synaptotagmin, both synaptotagmin and synaptophysin, and lines expressing all three synaptic vesicle proteins. In all cases, the proteins maintained their distinct compartmentalizations, were not found in the same organelle, and did not created synaptic vesicle-like structures. These results have important implications for models of synaptic vesicle biogenesis.
MeSH Terms
Animals
Brain/metabolism
CHO Cells
Calcium-Binding Proteins
Centrifugation, Density Gradient
Cricetinae
Drosophila/metabolism
Fibroblasts/metabolism
Fluorescent Antibody Technique
Humans
Membrane Glycoproteins/analysis,biosynthesis,metabolism
Microscopy, Electron
Microscopy, Immunoelectron
Nerve Tissue Proteins/analysis,biosynthesis,metabolism
Neurotransmitter Agents/metabolism
Rats
Subcellular Fractions/metabolism,ultrastructure
Synaptic Vesicles/metabolism
Synaptophysin/analysis,biosynthesis,metabolism
Synaptotagmins
Transfection
Chemicals
Calcium-Binding Proteins
Membrane Glycoproteins
Nerve Tissue Proteins
Neurotransmitter Agents
Sv2a protein, rat
Synaptophysin
Synaptotagmins
SV2A protein, human
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Feany M B
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115.
Yee A G
Delvy M L
Buckley K M
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