Occurrence of Two Types of Secretory Vesicles in the Human Neuroblastoma SH‐SY5Y

: Western blot analysis showed that the human neuroblastoma SH‐SY5Y expresses the proteins synaptotagmin I, synaptobrevin, synapsin I, rab3a, syntaxin, SNAP‐25, NSF, α‐SNAP, and munc‐18, which have been implicated in the movement, docking, and fusion of vesicles during exocytosis from other neuroend...

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Published in:Journal of neurochemistry 1997-04, Vol.68 (4), p.1542-1552
Main Authors: Goodall, Anna R., Danks, Kirsty, Walker, John H., Ball, Stephen G., Vaughan, Peter F. T.
Format: Article
Language:English
Subjects:
Antibody Specificity
Biological and medical sciences
Calcium-Binding Proteins
Carrier Proteins - analysis
Carrier Proteins - immunology
Cell physiology
Chromogranins
Cytoplasmic Granules - chemistry
Cytoplasmic Granules - metabolism
Exocytosis
Fluorescent Antibody Technique, Indirect
Fundamental and applied biological sciences. Psychology
Humans
Immunoblotting
Large dense‐cored vesicles
Membrane Glycoproteins - analysis
Membrane Proteins - analysis
Membrane Proteins - immunology
Molecular and cellular biology
N-Ethylmaleimide-Sensitive Proteins
Nerve Tissue Proteins - analysis
Nerve Tissue Proteins - immunology
Neuroblastoma
Neuropeptides - analysis
Noradrenaline
Norepinephrine - analysis
Proteins - analysis
Qa-SNARE Proteins
Secretion. Exocytosis
SH‐SY5Y neuroblastoma cells
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
Subcellular Fractions - chemistry
Synaptosomal-Associated Protein 25
Synaptotagmins
Tritium
Tumor Cells, Cultured - chemistry
Tumor Cells, Cultured - metabolism
Tumor Cells, Cultured - ultrastructure
Vesicular proteins
Vesicular Transport Proteins
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Summary:: Western blot analysis showed that the human neuroblastoma SH‐SY5Y expresses the proteins synaptotagmin I, synaptobrevin, synapsin I, rab3a, syntaxin, SNAP‐25, NSF, α‐SNAP, and munc‐18, which have been implicated in the movement, docking, and fusion of vesicles during exocytosis from other neuroendocrine cells. The subcellular localization of secretogranins I and II, synaptotagmin I, neuropeptide Y, rab3a, synaptobrevin, synaptophysin, and syntaxin was investigated by immunofluorescence microscopy and revealed punctate staining patterns characteristic of secretory vesicles. The comigration of noradrenaline, secretogranin II, and dopamine‐β‐hydroxylase on sucrose‐D2O gradient fractions indicates the presence of a population of noradrenaline‐containing large dense‐cored vesicles (LDCVs). In addition, a lighter vesicle population is also present that does not appear to be noradrenergic and contains a 48‐kDa synaptophysin antigen absent from the large dense‐cored vesicles. Immunocytochemical experiments show that not all of the vesicles that express synaptotagmin I contain secretogranin II. Thus, our studies suggest that two types of vesicle are present in SH‐SY5Y cells, one of which, the LDCVs, contains noradrenaline. These findings confirm our previous studies suggesting that depolarization‐evoked release of noradrenaline from SH‐SY5Y occurs by LDCV exocytosis. This enhances the value of SH‐SY5Y as a cell line in which to study the mechanism by which noradrenaline release is regulated.
ISSN:0022-3042
1471-4159
DOI:10.1046/j.1471-4159.1997.68041542.x