Effect of adenosine 3',5'-cyclic monophosphate on volume and cytoskeleton of MDCK cells.
Effect of adenosine 3',5'-cyclic monophosphate on volume and cytoskeleton of MDCK cells.
复制标题
腺苷3,5-环单磷酸对MDCK细胞体积和细胞骨架的影响。
作者：
J. Mills;M. Lubin
我们研究了N6，2 ′-O-二丁酰腺苷3 ′，5 ′-环一磷酸（DBcAMP）对Madin-Darby犬肾（MDCK）细胞融合培养物的体积和细胞骨架的影响。90分钟暴露于1 mM DBcAMP导致体积减少20%，如通过[14 C]-尿素水空间测量的。暴露于异丁基甲基黄嘌呤（IBMX，0.1 mM）的细胞体积减少了24%。在对照培养F-肌动蛋白，发现通过与硝基苯并恶二唑-phallacidin染色，在纤维的细胞的基础上，在交界区作为一个环带，并在顶端的细胞表面作为斑驳的荧光。通过间接免疫荧光显示，在整个细胞中可见密集的微管。暴露于DBcAMP 90分钟，导致F-肌动蛋白纤维的变化成致密的束附近的细胞的周边。当细胞暴露于IBMX时，这种效果甚至更加惊人。细胞松弛素B破坏F-肌动蛋白，并导致类似于DBcAMP的体积减少。DBcAMP和IBMX均不影响微管的分布。此外，秋水仙素，完全破坏微管，没有改变MDCK细胞体积。结果表明，DBcAMP和F-actin在MDCK细胞的体积控制中起作用。
We examined the effect of N6,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate (DBcAMP) on the volume and cytoskeleton of confluent cultures of Madin-Darby canine kidney (MDCK) cells. A 90-min exposure to 1 mM DBcAMP resulted in a 20% reduction in volume as measured by [14C]-urea water space. The volume in cells exposed to isobutylmethylxanthine (IBMX, 0.1 mM) was reduced by 24%. In control cultures F-actin, revealed by staining with nitrobenzoxadiazole-phallacidin, was found at the base of the cell as fibers, at the junctional region as a circumferential band, and on the apical cell surface as a mottled fluorescence. A dense pattern of microtubules, revealed by indirect immunofluorescence, was seen throughout the cell. Exposure to DBcAMP for 90 min resulted in a change of F-actin fibers into dense bundles near the periphery of the cell. This effect was even more striking when cells were exposed to IBMX. Cytochalasin B disrupted F-actin and resulted in a volume reduction similar to that in DBcAMP. Neither DBcAMP nor IBMX affected the distribution of microtubules. Moreover, colchicine, which completely disrupted the microtubules, did not change MDCK cell volume. The results suggest that DBcAMP and F-actin play a role in volume control in MDCK cells.