Actin localization in fixed dividing cells stained with fluorescent heavy meromyosin.
Actin localization in fixed dividing cells stained with fluorescent heavy meromyosin.
复制标题
肌动蛋白在固定分裂细胞中的定位，用荧光重粒肌球蛋白染色。
DOI：
10.1016/0014-4827(78)90030-7
复制
发表时间：
1978
影响因子：
3.7
通讯作者：
T. Pollard
中科院分区：
文献类型：
--
作者：
I. Herman;T. Pollard
我们重复了Sanger的[1]实验，用荧光重肌球蛋白（HMM）染色分裂细胞，在技术上做了一些重要的修改，并证实了他的一些观察结果，但不是全部。最重要的是，我们证实荧光- hmm集中在有丝分裂纺锤体中，即使在染色前用福尔马林固定的细胞中也是如此。这些实验中使用的荧光hmm通过离子交换色谱纯化，这一步消除了所有可检测的非特异性染色。荧光-HMM染色特异性的对照是与未标记的HMM竞争，以及肌动蛋白-荧光-HMM与焦磷酸镁相互作用的化学抑制。我们的荧光- hmm染色模式在三个方面与桑格的不同。我们没有观察到着丝点染色，也没有在卵裂沟中染色，而我们发现荧光- hmm在后期集中在区间。
We have repeated Sanger's [1] experiments on the staining of dividing cells with fluorescent-heavy meromyosin (HMM), with some important modifications in technique, and confirmed some, but not all, of his observations. Most importantly, we confirmed that fluorescent-HMM is concentrated in the mitotic spindle, even in cells that are fixed with formalin prior to staining. The fluorescent-HMM used in these experiments was purified by ion exchange chromatography, a step which eliminates all detectable non-specific staining. Controls for the specificity of the fluorescent-HMM staining were competition with unlabeled HMM and chemical inhibition of actin-fluorescent-HMM interaction with Mg-pyrophosphate. Our fluorescent-HMM staining patterns differed from Sanger's in three ways. We observed no staining of kinetochores and no concentration of staining in the cleavage furrow, while we found that fluorescent-HMM was concentrated in the interzone during anaphase.