Bashford C L, Chance B, Lloyd D, Poole R K
Biophys J. 1980 Jan;29(1):1-11. doi: 10.1016/S0006-3495(80)85114-9.
The redox state of the mitochondria of Acanthamoeba castellanii and Schizosaccharomyces pombe was assessed with a flying-spot fluorometer (Chance et al. 1978. Am. J. Physiol. 235:H 809) that provides excitation appropriate for oxidized flavoprotein or reduced pyridine nucleotide. Fluorescence signals could be resolved from the thin films of cultures that were only one cell deep. In both organisms anoxia was associated with an increased pyridine nucleotide and decreased flavoprotein fluorescence. The addition of mitochondrial uncoupling agents increased the flavoprotein fluorescence and the fluorometer was able to resolve uncoupler-sensitive and uncoupler-insensitive fractions of S. pombe cultures. In both synchronous and asynchronous cultures of A. castellanii and S. pombe the mitochondrial redox state oscillates with a period of 4.5 +/- 1.0 min. Oscillations with much longer period, of the order of an hour, are observed in synchronous cultures and these oscillations correlate with similar oscillations in respiratory rate, uncoupler sensitivity, and adenine nucleotide pool sizes. The results are consistent with the hypothesis that synchronous cultures of A. castellanii and S. pombe oscillate between the ADP-limited (state 4) and ADP-sufficient (state 3) respiratory states, i.e., exhibit in vivo respiratory control.
用飞点荧光计(Chance等人,1978年。《美国生理学杂志》235卷:H809)评估了卡氏棘阿米巴和粟酒裂殖酵母线粒体的氧化还原状态,该荧光计能提供适合氧化黄素蛋白或还原吡啶核苷酸的激发光。荧光信号可以从仅一个细胞厚度的培养薄膜中分辨出来。在这两种生物体中,缺氧都与吡啶核苷酸增加和黄素蛋白荧光减少有关。添加线粒体解偶联剂会增加黄素蛋白荧光,并且该荧光计能够分辨粟酒裂殖酵母培养物中对解偶联剂敏感和不敏感的部分。在卡氏棘阿米巴和粟酒裂殖酵母的同步和非同步培养物中,线粒体氧化还原状态都以4.5±1.0分钟的周期振荡。在同步培养物中观察到周期长得多(约一小时)的振荡,并且这些振荡与呼吸速率、解偶联剂敏感性和腺嘌呤核苷酸池大小的类似振荡相关。这些结果与以下假设一致,即卡氏棘阿米巴和粟酒裂殖酵母的同步培养物在ADP限制(状态4)和ADP充足(状态3)的呼吸状态之间振荡,即在体内表现出呼吸控制。