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RNA Docking and Local Translation Regulate Site-Specific Axon Remodeling In Vivo.

Neuron (2017-08-07)
Hovy Ho-Wai Wong, Julie Qiaojin Lin, Florian Ströhl, Cláudio Gouveia Roque, Jean-Michel Cioni, Roberta Cagnetta, Benita Turner-Bridger, Romain F Laine, William A Harris, Clemens F Kaminski, Christine E Holt
要旨

Nascent proteins can be positioned rapidly at precise subcellular locations by local protein synthesis (LPS) to facilitate localized growth responses. Axon arbor architecture, a major determinant of synaptic connectivity, is shaped by localized growth responses, but it is unknown whether LPS influences these responses in vivo. Using high-resolution live imaging, we examined the spatiotemporal dynamics of RNA and LPS in retinal axons during arborization in vivo. Endogenous RNA tracking reveals that RNA granules dock at sites of branch emergence and invade stabilized branches. Live translation reporter analysis reveals that de novo β-actin hotspots colocalize with docked RNA granules at the bases and tips of new branches. Inhibition of axonal β-actin mRNA translation disrupts arbor dynamics primarily by reducing new branch emergence and leads to impoverished terminal arbors. The results demonstrate a requirement for LPS in building arbor complexity and suggest a key role for pre-synaptic LPS in assembling neural circuits.

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Sigma-Aldrich
抗ピューロマイシン、クローン12D10、Alexa Fluor 488コンジュゲート抗体, clone 12D10, 0.5 mg/mL, from mouse
Sigma-Aldrich
3-マレイミド安息香酸 N-ヒドロキシスクシンイミドエステル, crystalline
Sigma-Aldrich
抗β-カテニン ウサギ宿主抗体, whole antiserum
Sigma-Aldrich
1,2-ジリノレオイル-3-パルミトイル-rac-グリセリン, ≥95% (TLC), liquid