Neural plasticity requires protein synthesis, but the identity of newly synthesized proteins generated in response to plasticity-inducing stimuli remains unclear. We used in vivo bio-orthogonal noncanonical amino acid tagging (BONCAT) with the methionine analog azidohomoalanine (AHA) combined with the multidimensional protein identification technique (MudPIT) to identify proteins that are synthesized in the tadpole brain over 24 hr. We induced conditioning-dependent plasticity of visual avoidance behavior, which required N-methyl-D-aspartate (NMDA) and Ca(2+)-permeable alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, alphaCaMKII, and rapid protein synthesis. Combining BONCAT with western blots revealed that proteins including alphaCaMKII, MEK1, CPEB, and GAD65 are synthesized during conditioning. Acute synthesis of CPEB during conditioning is required for behavioral plasticity as well as conditioning-induced synaptic and structural plasticity in the tectal circuit. We outline a signaling pathway that regulates protein-synthesis-dependent behavioral plasticity in intact animals, identify newly synthesized proteins induced by visual experience, and demonstrate a requirement for acute synthesis of CPEB in plasticity.
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Azidohomoalanine, MudPIT, Xenopus
Principal Investigators: (in alphabetical order) |
Hollis T. Cline, The Scripps Research Institute, USA |
Submitting User: | dmcclat |
Wanhua Shen, Han-Hsuan Liu, Lucio Schiapparelli, Daniel McClatchy, Hai-yan He, John R.Yates III, Hollis T.Cline.
Acute synthesis of CPEB is required for plasticity of visual avoidance behavior in Xenopus.
Cell Reports 6 , 737–747, February 27, 2014.
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