Synaptosomal cytoskeleton visualized by whole mount electron microscopy |
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Authors: | Tatsuo Suzuki Toshihiro Fujii Ryo Tanaka |
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Affiliation: | 1. Department of Biochemistry, Nagoya City University Medical School, Mizuho-ku, Nagoya 467, Japan;2. Department of Functional Polymer Science, Faculty of Textile Science and Technology, Shinshu University, Ueda 386, Japan |
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Abstract: | Organization of synaptosomal cytoskeleton was reproducibly visualized by the technique of whole mount electron microscopy. Synaptosomes from rat cerebrums were immobilized on the formvar membrane of the electron microscopic grid, partly solubilized by detergents of various kinds, and treated with chemicals to reveal cytoskeletons and their characteristics. Synaptosomal cytoskeletons consisted of three types: (1) pre-synaptic fiber network structure whose composite fiber was 15–20 nm in diameter and formed 60–100 nm circular rings. The rings had small particles inside and were organized into three-dimensional networks. The pre-synaptic network was different from the Triton-unextractable structure of mitochondria. (2) Post-synaptic fiber aggregate was constructed of 10-nm filaments that were typically visualized as deoxycholate- or N-lauroyl sarcosinate-unextractable cytoskeletons. The aggregate was a major structure in the Triton-unextractable cytoskeleton of synaptic plasma membrane (more than 95%). (3) Fiber connecting individual clusters of synaptosomal cytoskeletons which was probably an artifactual product formed during and after synaptosomal isolation. Existence of actin was indicated both in pre- and post-synaptic cytoplasm. |
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Keywords: | EDTA ethylenediaminetetraacetic acid EGTA HEPES mPSDp major postsynaptic density protein PHEM PIPES-HEPES-EGTA-Mg PIPES PSD post-synaptic density SJ synaptic junction SPM synaptic plasma membrane |
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