title: Glutamate-induced RNA localization and translation in neurons.
abstract: Localization of mRNA is required for protein synthesis to occur within discrete intracellular compartments. Neurons represent an ideal system for studying the precision of mRNA trafficking because of their polarized structure and the need for synapse-specific targeting. To investigate this targeting, we derived a quantitative and analytical approach. Dendritic spines were stimulated by glutamate uncaging at a diffraction-limited spot, and the localization of single β-actin mRNAs was measured in space and time. Localization required NMDA receptor activity, a dynamic actin cytoskeleton, and the transacting RNA-binding protein, Zipcode-binding protein 1 (ZBP1). The ability of the mRNA to direct newly synthesized proteins to the site of localization was evaluated using a Halo-actin reporter so that RNA and protein were detected simultaneously. Newly synthesized Halo-actin was enriched at the site of stimulation, required NMDA receptor activity, and localized preferentially at the periphery of spines. This work demonstrates that synaptic activity can induce mRNA localization and local translation of β-actin where the new actin participates in stabilizing the expanding synapse in dendritic spines.
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.|Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461.
authors: Young J
authors: YJ
authors: Yoon
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.|Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461.
authors: Bin
authors: B
authors: Wu
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.|Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Adina R
authors: AR
authors: Buxbaum
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors: Sulagna
authors: S
authors: Das
authors: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Albert
authors: A
authors: Tsai
authors: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Brian P
authors: BP
authors: English
authors: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Jonathan B
authors: JB
authors: Grimm
authors: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Luke D
authors: LD
authors: Lavis
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461; robert.singer@einstein.yu.edu.|Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461.|Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461.|Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors: Robert H
authors: 0000-0002-6725-0093
authors: RH
authors: Singer
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002465:Cell Movement; D003593:Cytoplasm / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism; D049229:Dendritic Spines / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D034443:RNA Transport / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D013569:Synapses / Q000378:metabolism
chemical_list: D000199:Actins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid
keywords: HaloTag; RNA localization; glutamate uncaging; single molecule; β-actin