INTRODUCTION: The proper dendritic branching is a highly regulated process. Among its regulators are membrane proteins internalized via clathrin-mediated endocytosis. AP2 adaptor complex is a key player in this process, but its role in mammalian dendritogenesis has not yet been tested. AIM(S): The aim of this study was to find how AP2 complex contributes to shaping dendritic tree of developing hippocampal neurons. METHOD(S): To study role of AP2 complex in dendritic arborization we used primary hippocampal neurons expressing AP2b1 (b-adaptin) shRNA alone or in combination of functional rescue constructs (i.a. GluA2, S6K1ca). The effect was also tested in vivo by lentiviral injections to newborn rats. Upon b-adaptin knockdown, we tested GluA2 trafficking via internalization assay and GluA2 level by Western blot and immunochemistry. GluA2 degradation and mTOR dependent biosynthesis were investigated by e.g. cycloheximide or rapamycin treatment. RESULTS: We showed that knockdown of b-adaptin led to reduction in dendritic arbors of developing hippocampal neurons in vitro and in vivo. The knockdown of AP2 also led to decreased level of GluA2, what is a result of impaired mTOR dependent GluA2 biosynthesis. However, the overexpression of functional GluA2 or restoration of mTOR activity rescued this effect. CONCLUSIONS: AP2 adaptor complex regulates the dendritogenesis of mammalian neurons via mTOR dependent GluA2 biosynthesis. FINANCIAL SUPPORT: This work has been financed by National Research Centre grant no. 2011/03/B/NZ3/01970.
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