Effects of ionophores and metabolic inhibitors on the mitochondrial membrane potential within isolated hepatocytes as measured with the safranine method.
Effects of ionophores and metabolic inhibitors on the mitochondrial membrane potential within isolated hepatocytes as measured with the safranine method.
复制标题
用番红法测量离子载体和代谢抑制剂对离体肝细胞内线粒体膜电位的影响。
DOI：
10.1042/bj1920183
复制
发表时间：
1980
期刊：
影响因子：
--
通讯作者：
J. Järvisalo
中科院分区：
文献类型：
--
作者：
K. Åkerman;J. Järvisalo
缬氨霉素或KCN与寡霉素联用后，在有番红存在的肝细胞悬液中缓慢出现一个差光谱，其吸收峰位于526 nm处。当细胞在37 ℃下在含有番红的培养基中孵育时，在波长对524-484 nm处发生吸光度的缓慢降低。在将细胞添加到含有番红的培养基后20-30分钟内完成吸光度的变化。此时，外部介质的番红浓度显著降低。番红信号被缬氨霉素、羰基氰化物对三氟甲氧基苯腙或KCN与寡霉素的组合完全逆转。这些处理对细胞ATP浓度或跨质膜的36 Cl-平衡电位没有任何直接影响。在碘乙酸存在下，在加入KCN后，可诱导痕量的缓慢逆转，但单独使用寡霉素则不行。鱼藤酮，与寡霉素组合，不逆转番红信号，除非当KF和碘乙酸都存在时，在这种情况下，可以看到一个缓慢的逆转。随后加入杜醌使信号恢复到与单独鱼藤酮存在时相同的水平。结果表明，在分离的肝细胞的存在下，番红的光谱响应是一个缓慢的渗透番红到细胞内线粒体，番红分子的聚集发生作为一个响应的线粒体膜电位的结果。
A difference spectrum with a peak of absorbance at 526nm appears slowly upon addition of valinomycin or KCN in combination with oligomycin to a hepatocyte suspension in the presence of safranine. When the cells are incubated at 37 degrees C in a medium containing safranine, a slow decrease in the absorbance occurs at the wavelength pair 524-484 nm. The change in absorbance is completed within 20-30 min after additions of cells to a medium containing safranine. At this time the safranine concentration of the outer medium is considerably decreased. The safranine signal is completely reversed by valinomycin, carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone or KCN in combination with oligomycin. None of these treatments have any immediate effect on cellular ATP concentrations or the 36Cl- equilibrium potential across the plasma membrane. In the presence of iodoacetate a slow reversal of the trace can be induced upon addition of KCN, but not of oligomycin alone. Rotenone, in combination with oligomycin, does not reverse the safranine signal except when both KF and iodoacetate are present, in which case a slow reversal is seen. A subsequent addition of duroquinone brings back the signal to the same level as in the presence of rotenone alone. The results indicate that the spectral response of safranine in the presence of isolated hepatocytes is a result of a slow penetration of safranine into intracellular mitochondria, where aggregation of safranine molecules occurs as a response to the mitochondrial membrane potential.