一种使用哺乳动物组织培养细胞研究胞质分裂的通透细胞模型。
A permeabilized cell model for studying cytokinesis using mammalian tissue culture cells.
作者信息
Cande W Z
出版信息
J Cell Biol. 1980 Nov;87(2 Pt 1):326-35. doi: 10.1083/jcb.87.2.326.
Abstract
PtK1 cells lysed late in cell division in a medium containing the nonionic detergent Brij 58 and polyethylene glycol with continue to undergo cleavage after lysis. Maintenance of cleavage after lysis is dependent on the composition of the lysis medium; the pH must be around neutrality, MgATP must be present, and the free Ca++ concentration should be 1 microM for optimal constriction to occur. Cleavage can be stopped and reinitiated by raising and lowering the Ca++ levels in the lysis medium. Cleavage in the permeabilized cell is blocked by addition of phalloidin, cytochalasin B, and N-ethylmaleimide-modified myosin subfragment-1 to the lysis medium. This represents the first cell model system for studying cleavage since the pioneering studies of Hoffman-Berling in 1954.
摘要
PtK1细胞在含有非离子去污剂Brij 58和聚乙二醇的培养基中于细胞分裂后期裂解后仍会继续进行卵裂。裂解后卵裂的维持取决于裂解培养基的成分；pH必须接近中性，必须存在MgATP，并且游离Ca++浓度应为1微摩尔才能发生最佳收缩。通过提高和降低裂解培养基中的Ca++水平，可以停止并重新启动卵裂。向裂解培养基中添加鬼笔环肽、细胞松弛素B和N-乙基马来酰亚胺修饰的肌球蛋白亚片段-1可阻断通透细胞中的卵裂。自1954年霍夫曼-伯林的开创性研究以来，这是第一个用于研究卵裂的细胞模型系统。
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