INCREASED MITOCHONDRIAL UPTAKE OF RHODAMINE-123 DURING LYMPHOCYTE STIMULATION
INCREASED MITOCHONDRIAL UPTAKE OF RHODAMINE-123 DURING LYMPHOCYTE STIMULATION
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DOI：
10.1073/pnas.78.4.2383
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
MELAMED, MR
中科院分区：
文献类型：
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作者：
DARZYNKIEWICZ, Z;STAIANOCOICO, L;MELAMED, MR
带正电的罗丹明类似物罗丹明123特异性地在活细胞的线粒体中积累。在植物血凝素刺激下，个体[人]淋巴细胞发生成母转化，用流式细胞术检测罗丹明123的摄取情况。在淋巴细胞刺激期间，观察到细胞积累罗丹明123的能力增加了几倍。在细胞刺激的第3天，最大的染料摄取与DNA合成（S期最大细胞数）和有丝分裂活性的高峰时间一致。在受刺激的淋巴细胞之间存在很大的细胞间差异，与未受刺激的淋巴细胞相比，一些细胞的荧光增加了15倍。罗丹明123摄取的增加也与细胞RNA含量的增加有关；染料摄取和细胞大小（通过光散射测量）之间的相关性不太明显。通过紫外显微镜观察，在胚胎形成过程中，染料摄取的增加在很大程度上是由于线粒体/细胞数量的增加。不能排除受刺激细胞中每个线粒体或每单位线粒体膜中罗丹明123结合的额外增加。罗丹明123可作为一种线粒体探针，用于区分循环细胞和静止细胞，并可用于分选功能不同的细胞亚群。
The positively charged rhodamine analog rhodamine 123 accumulates specifically in the mitochondria of living cells. The uptake of rhodamine 123 by individual [human] lymphocytes undergoing blastogenic transformation in cultures stimulated by phytohemagglutinin was measured by flow cytometry. A several-fold increase in cell ability to accumulate rhodamine 123 was observed during lymphocyte stimulation. Maximal dye uptake, seen on the 3rd day of cell stimulation, coincided in time with the peak of DNA synthesis (maximal number of cells in the S phase) and mitotic activity. A large intercellular variation among stimulated lymphocytes, with some cells having fluorescence increased as much as 15 times in comparison with nonstimulated lymphocytes, was observed. The increased uptake of rhodamine 123 also correlated with the increase in cellular RNA content; the correlation between the dye uptake and cell size (measured by light scatter) was less apparent. As observed by UV microscopy, the increased dye uptake during the blastogenesis was due, to a large extent, to an increase in number of mitochondria/cell. An additional increase in rhodamine 123 binding per mitochondrion or per unit of mitochondrial membrane in stimulated cells could not be excluded. Rhodamine 123 may be used as a supravital mitochondrial probe, discriminating between cycling and quiescent cells and having application in sorting functionally distinct cell subpopulations.