title: Distribution of synaptosomal-associated protein 25 in nerve growth cones and reduction of neurite outgrowth by botulinum neurotoxin A without altering growth cone morphology in dorsal root ganglion neurons and PC-12 cells.
abstract: Synaptosomal-associated protein 25 has been regarded as one of the target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptors essential for exocytosis of vesicles in synapses. We have previously reported that cleavage of syntaxin, which is another target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptor, with botulinum neurotoxin C1 resulted in inhibition of neurite extension and morphological changes including growth cone collapse and large vacuole formation. As an attempt to explore the mechanism of growth cone extension, we examined the ultrastructural localization of synaptosomal-associated protein 25 in growth cones with or without treatment of botulinum neurotoxin A, which cleaves synaptosomal-associated protein 25. In dorsal root ganglion neurons, light microscopy demonstrated synaptosomal-associated protein 25 immunoreactivity throughout the neurons, including the cell bodies, neurites and growth cones. Using electron microscopy, gold signals immunoreactive for synaptosomal-associated protein 25 were identified diffusely in the cytoplasm of the growth cones. In contrast, in PC-12 cells, a large number of gold signals were localized on the plasma membranes. High levels of signal were also found in the cytoplasm in the central region of the growth cones. We also confirmed that botulinum neurotoxin A treatment reduced neurite extension by about 50%. However, both in dorsal root ganglion neurons and in PC-12 cells we found no differences in the ultrastructure nor in the localization of synaptosomal-associated protein 25 between growth cones with and without toxin treatment. These results indicate that cleavage of synaptosomal-associated protein 25 inhibits growth cone extension in a manner different than that of syntaxin cleavage. The results of this study suggest the possibility that synaptosomal-associated protein 25 is involved in growth cone extension through a process independent of vesicle fusion.
authors.affiliation: Department of Anatomy and Neurobiology, Kyoto University Graduate School of Medicine, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Morihara
authors.forename: A
authors.initials: A
authors.lastname: Mizoguchi
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: S
authors.initials: S
authors.lastname: Kozaki
authors.forename: T
authors.initials: T
authors.lastname: Tsujihara
authors.forename: S
authors.initials: S
authors.lastname: Kawano
authors.forename: M
authors.initials: M
authors.lastname: Shirasu
authors.forename: T
authors.initials: T
authors.lastname: Ohmukai
authors.forename: M
authors.initials: M
authors.lastname: Kitada
authors.forename: K
authors.initials: K
authors.lastname: Kimura
authors.forename: S
authors.initials: S
authors.lastname: Okajima
authors.forename: K
authors.initials: K
authors.lastname: Tamai
authors.forename: Y
authors.initials: Y
authors.lastname: Hirasawa
authors.forename: C
authors.initials: C
authors.lastname: Ide
mesh_terms: D000818:Animals; D001369:Axons / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D019274:Botulinum Toxins, Type A / Q000494:pharmacology*; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology*; D008565:Membrane Proteins; D051379:Mice; D008815:Mice, Inbred Strains; D016253:Microscopy, Immunoelectron; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D016501:Neurites / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D016716:PC12 Cells; D051381:Rats; D050825:Synaptosomal-Associated Protein 25
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C494209:Snap25 protein, mouse; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25; D019274:Botulinum Toxins, Type A