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KMID : 0390320130230010069
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Chungbuk Medical Journal
2013 Volume.23 No. 1 p.69 ~ p.77
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Development of the Method to Measure Protein Synthesis in Neuronal Processes
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Jeong Hyun-Ju
Kim Hyong-Kyu
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Abstract
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Purpose : Local protein synthesis in neuronal processes has a key role in cellular responses to extracellular stimuli, such as synaptic plasticity. As there is not any biochemical method for measuring only local translation till now, the study of that has been restricted. In this study, we tried to develope a new method to measure local translation only and to evaluate it. Materials and Methods : Rat hippocampal neurons were incubated on the membrane insert containing 8 μm-wide pores for neuronal processes to be physically isolated from the cell bodies. A reporter system was developed by using untranslated regions (UTR) of Kv4.2 mRNA and luciferase gene, and then it was used in isolation of RNA-binding proteins which were involved in local translation of Kv4.2 potassium channel. The role of isolated RNA-binding protein in local protein synthesis was examined by taking advantage of the culture system. Results : We developed a new culture system to measure local protein synthesis in neuronal processes. We isolated a RNA-binding protein involved in local translation of Kv4.2 potassium channel and confirmed it by using the measuring system. Conclusion : We established a measuring system for local protein synthesis.
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KEYWORD
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Local protein synthesis, Neuronal process, Reporter, RNA-binding protein
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