ANALYSIS OF CELL-DIVISION USING FLUORESCENTLY LABELED ACTIN AND MYOSIN IN LIVING PTK2-CELLS
ANALYSIS OF CELL-DIVISION USING FLUORESCENTLY LABELED ACTIN AND MYOSIN IN LIVING PTK2-CELLS
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DOI：
10.1002/cm.970140207
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发表时间：
1989-01-01
影响因子：
--
通讯作者：
SANGER, JW
中科院分区：
文献类型：
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作者：
SANGER, JM;MITTAL, B;SANGER, JW
肌动蛋白和肌球蛋白的轻链用荧光染料标记，并注射到间期PtK2细胞中，以研究注射的细胞随后进入有丝分裂和分裂时发生的肌动蛋白和肌球蛋白分布的变化。第一个变化发生时，应力纤维在前期细胞开始解体。在这个从细胞中心开始的过程中，单个纤维缩短，在少数纤维中，可以看到相邻的荧光肌球蛋白带移动得更近。在大多数细胞中，应力纤维解体是由中期完成的，导致在整个细胞质中的荧光蛋白的扩散分布与最大的浓度存在于有丝分裂纺锤体。肌动蛋白和肌球蛋白浓度的第一个证据发生在分裂后期，就在开沟可以检测到。最初，荧光强度和荧光环的宽度随着环收缩而增加。在不对称定位的有丝分裂纺锤体的细胞中，蛋白质浓度和开沟首先在最接近有丝分裂纺锤体赤道的皮质区域明显。随着胞质分裂在这种不对称分裂的细胞中进行，荧光肌动蛋白和肌球蛋白出现在细胞的另一侧，就在那里可以看到开沟活动之前。在胞质分裂结束时，肌球蛋白和肌动蛋白集中在中间体的膜下，随后在中间体的两端组织成两个环。
Actin and the light chains of myosin were labeled with fluorescent dyes and injected into interphase PtK2 cells in order to study the changes in distribution of actin and myosin that occurred when the injected cells subsequently entered mitosis and divided. The first changes occurred when stress fibers in prophase cells began to disassemble. During this process, which began in the center of the cell, individual fibers shortened, and in a few fibers, adjacent bands of fluorescent myosin could be seen to move closer together. In most cells, stress fiber disassembly was complete by metaphase, resulting in a diffuse distribution of the fluorescent proteins throughout the cytoplasm with the greatest concentration present in the mitotic spindle. The first evidence of actin and myosin concentration in a cleavage ring occurred at late anaphase, just before furrowing could be detected. Initially, the intensity of fluorescence and the width of the fluorescent ring increased as the ring constricted. In cells with asymmetrically positioned mitotic spindles, both protein concentration and furrowing were first evident in the cortical regions closest to the equator of the mitotic spindle. As cytokinesis progressed in such asymmetrically dividing cells, fluorescent actin and myosin appeared at the opposite side of the cell just before furrowing activity could be seen there. At the end of cytokinesis, myosin and actin were concentrated beneath the membrane of the midbody and subsequently became organized in two rings at either end of the midbody.