大鼠肾上腺嗜铬细胞的动作电位与量子化分泌

We have developed a new method, in which we use single real action potential waveform as basic unit to encode arbitrary action potentials represented as 4-parameter functions F (n,m,f,d). We input certain F (n,m,f,d) into voltage clamped Rat Adrenal Chromaffin cells (RACC) and record the cell secretion response. The method was applied to study of DRG neurons and resulted in the publication of the first Nature Neuroscience paper of China. With this method we studied action potential's regulation on RACC' s secretion, and discovered that the number of action potential bursts has important function on secretion, when the number and frequency of Action potential remain unaltered. The quantity of secretion when m=4 is double as much as that when m=2.

本研究利水蛭素C端12肽基因片段为连接肽基因融合方法构建Annexin V-scu-pA（144-411）融合基因，构建了大肠杆菌表达质粒pET-28-UKA，在大肠杆菌BL21（DE3）中用IPTG诱导表达，表达产物在复性后具有明显的纤溶活性。我们又用Bac-Bac昆虫杆状病毒表达系统在昆虫细胞sf中表达了融合蛋白基因Annexin V-scu-pA（144-411），获得了较高的表达量。活性研究表明，表达产物不仅具有低分子量尿激酶的纤溶活性，而且具有Annexin V的抗凝活性。研究还表明，用酵母表达系统来工业化获取此融合蛋白，仍有相当难度。本项目研究结果已申请国家发明专利一件，有望开发成具有临床应用价值的新物。