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曲拉通X-100对分离的肝细胞核的核膜的影响。

The influence of Triton X-100 on the nuclear envelope of the isolated liver cell nuclei.

作者信息

Frederiks W M, James J, Arnouts C, Broekhoven S, Morreau J

出版信息

Cytobiologie. 1978 Dec;18(2):254-71.

PMID:215470
Abstract

In order to obtain more precise information on an eventual presence of extra-membranous lipids in the interior of the nucleus, the effects of Triton X-100 on the lipid content and ultrastructure of isolated rat liver nuclei was investigated. Enzyme markers (a.o. glucose-6-phosphatase) were used to control impurities of the nuclear fractions biochemically along with transmission electron microscopy and qualitative and quantitative light microscopy to check the condition of the nuclei obtained. Treatment of the nuclear fraction with increasing concentrations of Triton X-100 resulted in a decrease of the phospholipid content down to 25% at a Triton X-100/protein ratio of 0.4. A further decrease to 8% was measured at a ratio of 1.5. Electron microscopy of nuclei of the latter group showed nuclei containing outer membrane fragments in 2.5% of their surfaces. The composition of lipids extracted from a nuclear fraction appeared to be markedly changed after treatment with Triton X-100 with an increase of the percentage of neutral lipids and the phospholipids diphosphatidyl-glycerol and spingomyelin. From the chemical and morphological data obtained, the conclusion was drawn that a substantial part of the lipids remaining in the isolated nuclei after treatment with Triton X-100 is localized in both membranes of the nuclear envelope. It cannot however, be excluded that a small portion would be present in the interior of the nuclei.

摘要

为了获取关于细胞核内部是否存在膜外脂质的更精确信息,研究了Triton X - 100对分离的大鼠肝细胞核脂质含量和超微结构的影响。使用酶标记物(如葡萄糖 - 6 - 磷酸酶)通过生化方法控制核组分的杂质,同时利用透射电子显微镜以及定性和定量光学显微镜检查所获得细胞核的状况。用浓度不断增加的Triton X - 100处理核组分,当Triton X - 100/蛋白质比例为0.4时,磷脂含量降低至25%。当比例为1.5时,进一步降至8%。对后一组细胞核进行电子显微镜观察显示,其细胞核表面有2.5%含有外膜碎片。用Triton X - 100处理后,从核组分中提取的脂质组成明显改变,中性脂质以及磷脂二磷脂酰甘油和鞘磷脂的百分比增加。根据所获得的化学和形态学数据得出结论,用Triton X - 100处理后残留在分离细胞核中的大部分脂质定位于核膜的两层膜中。然而,不能排除细胞核内部存在一小部分脂质的可能性。

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