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newly-synthesized proteins are required for visual experience dependent behavioral plasticity in the developing brain

Newly-synthesized proteins are required for visual experienc...

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中国神经科学学会第十二届全国学术会议

作者： Xianjie Qi Zhaoyi Guo Lijun Zheng Juanmei Gao Yuan Liao Wanhua Shen Zhejiang Key Laboratory of Organ Development and Regeneration College of Life and Environmental Sciences Hangzhou Normal University

Neural circuit formation and plasticity are largely dependent on the protein synthesis in the central nervous system. We have shown that newly-synthesized proteins are necessary for the behavioral plasticity in the developing Xenopus tadpoles. The visual avoidance behavior is greatly improved in response to 4 hour visual conditioning at stage 47 tadpoles. In particular, the enhancement of the visual avoidance index is largely blocked by a protein synthesis inhibitor, indicating that newly-synthesized proteins are required for the behavioral plasticity. To determine the identity of the newly-synthesized proteins, the tectal brain was injected with a methionine analogue of Azidohomoalanin（AHA） and tectal brains were homogenized to perform click chemistry reaction and Western blot. We find that the newly-synthesized proteins are significantly increased in the visual conditioned tadpoles compared to the control tadpoles. The increment of newly-synthesized proteins are also observed by the experiment of Dot blot. To screen the identity of newly-synthesized proteins, we combined the technologies of biorthogonal noncanonical amino acid tagging（BONCAT）and the label-free mass spectrometry. We find that hundreds of newly-synthesized proteins are dramatically increased in the visual conditioned optic tecta. It is interesting to find that the synaptic proteins of Rab3 family members of Rab3 a, Rab3 b and Rab3 c are differentially regulated in response to visual conditioning. Rab3 b and Rab3 c are significantly increased, while Rab3 a was dramatically decreased in the visual stimulated tectal brains by Western blot. The functional role of Rab3 family members in the visual experience-dependent behavioral plasticity is to be determined in our following experiments. These data suggested that BONCAT combined with Mass spectrometry largely improves the sensitivity for identifying newlysynthesized proteins in the central nervous system. Rab3 family members may play differential roles in mediating visual experience-dependent behavio

关键词： click chemistry behavioral plasticity newly-synthesized proteins Rab3

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