2001 Fiscal Year Final Research Report Summary
Development and application ofa novel genetic strategy for visualization offunctional neural pathways with WGA-GFP fusion transgene
| Project/Area Number |
12480243
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | RIKEN |
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Principal Investigator |
YOSHIHARA Yoshihiko RIKEN, Lab. for Neurobioology of Synapse, Laboratory Head, シナプス分子機構研究チーム, チームリーダー(研究職) (20220717)
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| Co-Investigator(Kenkyū-buntansha) |
FUNAKI Atsuko RIKEN, Lab. For Newrobiology of Synapse, Technical Staff, シナプス分子機構研究チーム, テクニカルスタッフ(研究職)
SAIO Michiko RIKEN, Lab. For Neurobiology of Synapse, Technical Staff, シナプス分子機構研究チーム, テクニカルスタッフ(研究職)
MIYAWAKI Atsushi RIKEN, Lab. For Cell Function Dynamics, Laboratory Head, 細胞機能探索技術探索チーム, チームリーダー(研究職) (80251445)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Wheat Germ Agglutinin / Transsynaptic Tracer / Neural Network / Transgenic Mouse / Green Fluorescent Protein / Synapse / Visualization / Neuroanatomy |
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| Research Abstract |
Information transfer between neurons takes place at the synapse. The wiring patterns among various types of neurons via specific synaptic connections are the basis of functional logic employed by the brain for information processing. Thus, detailed knowledge of neuronal networks is essential for understanding the wide range of brain fhrlctions. We previously developed a powerfirl genetic strategy for visualization of specific neuronal pathways across a synapse by combining a neuroanatomical tracing method with transgenic and gene targeting technology. By introducing cDNA for a plant lectit, wheat germ agglutinin (WGA), as a transgene under the control of specific promoter elements, selective and ftinctional transsynaptic neural pathways could be visualized. To detect the WGA transgene product immunohistochemically with anti-WGA antibody, however, the animals must be sacrificed and the tedious and time-consuming procedures are needed. For more convenient and simple detection of transsynaptic reporter, we attempted to develop fusion transgenes consisting of WGA and various fluorescent proteins (green : GFP, cyan : CFP, yellow : YFP, red : DsRed). If such chimeric molecules could be efficiently transferred across synapses with bright fluorescence, it would become possible to visualize functional neural pathways in situ or ultimately in vivo and to combine this transsynaptic tracing technique with various physiological measurements. We made several ftision transgenes and found that these molecules show bright fluorescence in cultured neurons. We are now checking whether the fluorescent WGA can be transferred across a synapse or not.
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Research Products
(9 results)
