VAMP-2 promotes neurite elongation and SNAP-25A increases neurite sprouting in PC12 cells

• Published in: Neuroscience research Vol. 37; no. 4; pp. 265 - 275
• Main Authors: Shirasu, Mikihiro, Kimura, Kazushi, Kataoka, Masakazu, Takahashi, Masami, Okajima, Seiichiro, Kawaguchi, Saburo, Hirasawa, Yasusuke, Ide, Chizuka, Mizoguchi, Akira
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 01.08.2000
• Subjects: Animals; Antigens, Surface - genetics; Antigens, Surface - metabolism; Green fluorescent protein; Green Fluorescent Proteins; Immunoblotting; Luminescent Proteins - genetics; Membrane Proteins - genetics; Membrane Proteins - metabolism; Microscopy, Confocal; Microscopy, Immunoelectron; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neurite outgrowth; Neurites - physiology; PC12 Cells; Precipitin Tests; R-SNARE Proteins; Rats; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; SNAP-25A; Synaptosomal-Associated Protein 25; Syntaxin 1; Syntaxin1A; Transfection; VAMP-2; Transfection; VAMP-2; SNAP-25A; Syntaxin1A; Neurite outgrowth; Green fluorescent protein
• ISSN: 0168-0102; 1872-8111
• DOI: 10.1016/S0168-0102(00)00125-5

Recent studies suggest that the soluble N-ethylmaleimide-sensitive factor attached protein (SNAP) receptor (SNARE)-mediated membrane fusion system is involved in vesicle fusion in the plasma membrane that allows expansion for neurite elongation. There have been several reports analyzing the effects of neurite outgrowth by inhibition of SNAREs. In this study, we took the opposite approach by overexpressing green fluorescent protein (GFP)-fusion SNAREs, including VAMP-2, SNAP-25A, and syntaxin1A, in PC12 cells to investigate the role of SNAREs in the neurite outgrowth of PC12 cells. Neurite outgrowth analysis demonstrated that: (1) GFP-VAMP-2 increased the length of individual neurites, without changing the number of neurites per cell; (2) GFP-SNAP-25A increased the number of neurites per cell, with no change in the length of the individual neurites. In both cases, the total length of neurites per cell was increased; (3) GFP-syntaxin1A resulted in no significant change, either in neurite length, or in the number of neurites per cell. These findings suggest that when overexpressed in PC12 cells, VAMP-2 can promote neurite elongation, while SNAP-25A can stimulate neurite sprouting. On the other hand, overexpression of syntaxin1A neither promotes nor inhibits neurite outgrowth. Thus VAMP-2 and SNAP-25A play different roles in neurite elongation and sprouting.