Kindzelskii A L, Zhou M J, Haugland R P, Boxer L A, Petty H R
Department of Biological Sciences, Wayne State University, Detroit, Michigan 48202, USA.
Biophys J. 1998 Jan;74(1):90-7. doi: 10.1016/S0006-3495(98)77770-7.
To better understand the mechanism of leukocyte migration in complex environments, model extracellular matrices were prepared using gelatin, Hanks' solution, Bodipy-BSA (fluorescent upon proteolysis), and dihydrotetramethylrosamine or hydroethidine (fluorescent upon oxidation). Using quantitative microfluorometry, neutrophil-mediated extracellular pulses of reactive oxygen metabolites (ROMs) and pericellular proteolysis were periodically observed showing that these functions occur as quantal bursts. However, chronic granulomatous disease neutrophils, which do not produce ROMs, did not display ROM deposition. Matrices show an alternating pattern of green (proteolytic) and red (oxidative) fluorescence, indicating these functions are out of phase. Electric fields phase-matched with metabolic oscillations, which increase the amplitude of intracellular NAD(P)H oscillations, increase ROM deposition and pericellular proteolysis; this further supports the link between intracellular chemical oscillators and extracellular functions. This phase relationship may allow ROMs to inactivate protease inhibitors, followed by protease activation.
为了更好地理解白细胞在复杂环境中的迁移机制，使用明胶、汉克斯溶液、Bodipy-BSA（蛋白水解时发出荧光）以及二氢四甲基罗丹明或氢乙锭（氧化时发出荧光）制备了细胞外基质模型。通过定量显微荧光测定法，周期性观察到中性粒细胞介导的活性氧代谢产物（ROMs）的细胞外脉冲和细胞周围蛋白水解现象，表明这些功能以量子爆发的形式出现。然而，不产生ROMs的慢性肉芽肿病中性粒细胞并未显示出ROM沉积。基质呈现出绿色（蛋白水解）和红色（氧化）荧光交替的模式，表明这些功能不同步。与代谢振荡相位匹配的电场会增加细胞内NAD(P)H振荡的幅度，从而增加ROM沉积和细胞周围蛋白水解；这进一步支持了细胞内化学振荡器与细胞外功能之间的联系。这种相位关系可能使ROMs使蛋白酶抑制剂失活，随后激活蛋白酶。
