Application of SDS-FRL for Quantitative Localization of Brain Molecules
| Project/Area Number |
18500252
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neuroscience in general
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| Research Institution | National Institute for Physiological Sciences |
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Principal Investigator |
FUKAZAWA Yugo National Institute for Physiological Sciences, Cerebral Research, Assistant Professor (60343745)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,940,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | Anatomy / Synapse / Neuroscience / Receptor / Electron microscopy / 分子局在 |
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| Research Abstract |
One of the questions to be answered in the post-genomic era, at a time when most proteins constituting living organisms have been identified, is what particular protein species and amount of each species is expressed in a particular cell type and in their subcellular domains. Such information is indispensable for understanding mechanisms involved in living cells. For subcellular localization that requires nano-scale resolution, conventional immunoelectron microscopy (pre-and post-embedding immunolabeling) has been widely used. Although these techniques, especially with immunogold particles, have provided precise details of molecular localization in different subcellular domains, reliable quantification of immunoreactivity has often been hampered by low sensitivity and limited accessibility of antibodies to target molecules buried in tissues. In 1995, Fujimoto developed an epoch-making technique for localization of plasma membrane molecules, termed SDS-digested freeze-fracture replica labeling (SDS-FRL), by which molecules on the plasma membrane can be directly approached by antibodies and visualized in a two-dimensional manner with high sensitivity and high resolution. In this research project, I have applied some modifications onto this technique in order to establish quantitative localization method useful for neuroscience research using brain tissue. By combining outcomes from these efforts, I established and introduced current protocol of SDS-FRL in some original papers, which could be further modified for various purposes and other tissues to bring out the full potential of this powerful technique.
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Report
(3 results)
Research Products
(26 results)
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[Journal Article] SDS-digested Freeze-fracture replica labelling(SDS-FRL)2007
Author(s)
Fukazawa, Y., Masugi-Tokita, M., Tarusawa, E., Hagiwara, A., Shigemoto, R.
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Journal Title
In Handbook of Cryo-Preparation Methods for Electron Microscopy(Eds. Cavalier, A., Spehner, D., & Humbel, B.M.), CRC Press, Boca Raton, USA, 559-576
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] A CaV2.1 calcium channel mutation rocker reduces the number of postsynaptic AMPA receptors in parallel fiber-Purkinje cell synapses2006
Author(s)
Kodama, T., Itsukaichi-Nishida, Y., Fukazawa, Y., Wakamori, M., Miyata, M., Molnar, E., Mori, Y., Shigemoto, R., & Imoto, K.
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Journal Title
European Journal of Neuroscience 24
Pages: 2993-3007
Description
「研究成果報告書概要(和文)」より
Related Report
Peer Reviewed
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