TRYPAN BLUE AS A MARKER OF PLASMA-MEMBRANE PERMEABILITY IN ALLOXAN-TREATED MOUSE ISLET CELLS
TRYPAN BLUE AS A MARKER OF PLASMA-MEMBRANE PERMEABILITY IN ALLOXAN-TREATED MOUSE ISLET CELLS
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DOI：
10.1007/bf03349305
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发表时间：
1979-01-01
影响因子：
5.4
通讯作者：
TALJEDAL, IB
中科院分区：
文献类型：
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作者：
GRANKVIST, K;LERNMARK, A;TALJEDAL, IB
将非inbredob/ob小鼠的胰岛细胞悬液与台盼蓝孵育。显微镜光度法显示，表面上有活力的细胞完全排除了染料，而没有活力的细胞的细胞核通过饱和过程积累了台盼蓝。在含有0.1%或更少台盼蓝的介质中，细胞核与介质的染料梯度大于30：1。染料与核结合的表观亲和常数为3.1×104L/mol。白蛋白部分抑制了核染色。培养液中台盼蓝含量超过0.5%即为毒物。无白蛋白时，0.5或20 mmo1/L四氧嘧啶、1 mmo1/Ln-乙基马来酰亚胺或0.1 mmo1/L氯化汞对磺酸，而20 mmo1/L链脲佐菌素则不能增加0.1%台盼蓝染色胰岛细胞的频率。在四氧嘧啶或乙基马来酰亚胺处理的细胞中，细胞核的吸光度也增加，但在氯化汞-对磺酸处理的细胞中，核的吸光度没有增加。结论：四氧嘧啶可迅速增加小鼠β-细胞质膜通透性。四氧嘧啶的这种作用似乎比链脲佐菌素的任何这种作用都更剧烈。
Suspensions of pancreatic islet cells from noninbredob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant for nuclear binding of the dye was 3.1 × 104l/mol. Albumin partially inhibited the nuclear staining. More than 0.5% Trypan Blue in the medium was toxicper se. In the absence of albumin, 0.5 or 20 mmol/l alloxan, 1 mmol/lN-ethylmaleimide, or 0.1 mmol/l chloromercuribenzene-p-sulphonic acid, but not 20 mmol/l streptozotocin, increased the frequency of islet cells stained with 0.1% Trypan Blue. The absorbance of nuclei was also increased in cells treated with alloxan orN-ethylmaleimide, but not in those treated with chloromercuribenzene-p-sulphonic acid. It is concluded that alloxan rapidly increases the permeability of the plasma membrane in mouse β-cells. This action of alloxan appears to be more acute than any such effect of streptozotocin.