| Co-Investigator(Kenkyū-buntansha) |
TANAKA Satoshi Gunma University School of Medicine, Department of Neurobiology and Behavior, Research Associate, 医学部, 助手 (20272247)
SEKINO Yuko Gunma University School of Medicine, Department of Neurobiology and Behavior, Assistant Professor, 医学部, 講師 (70138866)
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| Research Abstract |
Drebrins are actin-binding proteins, which expression is closely related to spine morphology. Therefore we hypothesized that drebrin-family proteins play adaptor proteins of postsynaptic signal transmission from membrane receptor to actin cytoskeleton.
In the previous study, we established the primary culture of cortical neurons, in which neurons can develop and the dendritic spine seemed to be fully maturated. Using this culture systems, we inhibit the drebrin expression wit antisense-oligonucleotides, and demonstrated that activity-dependent accumulation of synaptic functional proteins were changed.
Next, we hypothesized that other drebrin-binding proteins than actin play an important role in postsynaptic signal transmission in the neuron. We performed the yeast two hybrid system for cloning the binding proteins to drebrin-family proteins (drebrin E, drebrin A, and SH3P7).
One of the clone, designated as DRAP1, encodes full length of a coda of a novel drebrin-binding protein. Then we performed its biochemical characterization and found its N-terminal half actually binds to the N-terminal region of a drebrin molecule. Next, using polypeptide of DRAP1 and DRAP1 fragment expressed by gene engineering technique, we raise polyclonal antiserum to DRAP1.
Further, we constructed expression vector which encode the GFP-DRAP1 fused protein. When we transformed fibroblasts with the expression vector, we observed GFP fluorescence in their nuclei.
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