Agarwal Hitesh K, Janicek Radoslav, Chi San-Hui, Perry Joseph W, Niggli Ernst, Ellis-Davies Graham C R
Department of Neuroscience, Mount Sinai School of Medicine , New York, New York 10029, United States.
Department of Physiology, University of Bern , Bern CH 3012, Switzerland.
J Am Chem Soc. 2016 Mar 23;138(11):3687-93. doi: 10.1021/jacs.5b11606. Epub 2016 Mar 14.
We have designed a nitroaromatic photochemical protecting group that absorbs visible light in the violet-blue range. The chromophore is a dinitro derivative of bisstyrylthiophene (or BIST) that absorbs light very effectively (ε440 = 66,000 M(-1) cm(-1) and two-photon cross section of 350 GM at 775 nm). We developed a "caged calcium" molecule by conjugation of BIST to a Ca(2+) chelator that upon laser flash photolysis rapidly releases Ca(2+) in <0.2 ms. Using the patch-clamp method the optical probe, loaded with Ca(2+), was delivered into acutely isolated mouse cardiac myocytes, where either one- and two-photon uncaging of Ca(2+) induced highly local or cell-wide physiological Ca(2+) signaling events.
我们设计了一种硝基芳香族光化学保护基团，它能吸收紫蓝色范围内的可见光。发色团是双苯乙烯基噻吩（或BIST）的二硝基衍生物，其吸光效果非常好（ε440 = 66,000 M(-1) cm(-1)，在775 nm处的双光子截面为350 GM）。我们通过将BIST与Ca(2+)螯合剂共轭，开发了一种“笼状钙”分子，在激光闪光光解时，该分子能在<0.2毫秒内迅速释放Ca(2+)。使用膜片钳方法，将装载有Ca(2+)的光学探针导入急性分离的小鼠心肌细胞中，在那里，Ca(2+)的单光子和双光子解笼会诱导高度局部或全细胞范围的生理性Ca(2+)信号事件。
