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Cholesterol supports the retinoic acid‐induced synaptic vesicle formation in differentiating human SH‐SY5Y neuroblastoma cells
Author(s) -
Sarkanen JerttaRiina,
Nykky Jonna,
Siikanen Jutta,
Selinummi Jyrki,
Ylikomi Timo,
Jalonen Tuula O.
Publication year - 2007
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2007.04676.x
Subject(s) - synaptic vesicle , synaptophysin , neurite , microbiology and biotechnology , biology , sh sy5y , vesicle , exocytosis , synaptic vesicle recycling , neuroblast , cell culture , chemistry , neuroblastoma , biochemistry , in vitro , immunology , membrane , genetics , immunohistochemistry , neurogenesis
Synaptic vesicle formation, vesicle activation and exo/endocytosis in the pre‐synaptic area are central steps in neuronal communication. The formation and localization of synaptic vesicles in human SH‐SY5Y neuroblastoma cells, differentiated with 12‐ o ‐tetradecanoyl‐phorbol‐13‐acetate, dibutyryl cyclic AMP, all‐ trans ‐retinoic acid (RA) and cholesterol, was studied by fluorescence microscopy and immunocytochemical methods. RA alone or together with cholesterol, produced significant neurite extension and formation of cell‐to‐cell contacts. Synaptic vesicle formation was followed by anti‐synaptophysin (SypI) and AM1‐43 staining. SypI was only weakly detected, mainly in cell somata, before 7 days in vitro , after which it was found in neurites. Depolarization of the differentiated cells with high potassium solution increased the number of fluorescent puncta, as well as SypI and AM1‐43 co‐localization. In addition to increase in the number of synaptic vesicles, RA and cholesterol also increased the number and distribution of lysosome‐associated membrane protein 2 labeled lysosomes. RA‐induced Golgi apparatus fragmentation was partly avoided by co‐treatment with cholesterol. The SH‐SY5Y neuroblastoma cell line, differentiated by RA and cholesterol and with good viability in culture, is a valuable tool for basic studies of neuronal metabolism, specifically as a model for dopaminergic neurons.