Melby E L, Prydz K, Olsnes S, Sandvig K
Institute for Cancer Research, Norwegian Radium Hospital, Montebello, Oslo.
J Cell Biochem. 1991 Nov;47(3):251-60. doi: 10.1002/jcb.240470311.
The effect of monensin on endocytosis, transcytosis, recycling and transport to the Golgi apparatus in filter-grown Madin-Darby canine kidney (MDCK) cells was investigated using 125I-labeled ricin as a marker for membrane transport, and horseradish peroxidase (HRP) as a marker for fluid phase transport. Monensin (10 microM) stimulated transcytosis of both markers about 3-fold in the basolateral to apical direction. Transcytosis of HRP in the opposite direction, apical to basolateral, was reduced to approximately 50% of the control by monensin, whereas that of ricin was slightly increased. Recycling of markers endocytosed from the apical surface was reduced in the presence of monensin and there was an increased accumulation of both ricin and HRP in the cells. Transport of ricin to the Golgi apparatus increased to the same extent as the increase in intracellular accumulation. No change in recycling or accumulation was observed with monensin when the markers were added basolaterally, but transport of ricin to the Golgi apparatus increased almost 3-fold. Our results indicate that basolateral to apical transcytosis is increased in the absence of low endosomal pH, and they suggest that apical to basolateral transcytosis of a membrane-bound marker (ricin) is affected by monensin differently from that of a fluid phase marker (HRP).
使用125I标记的蓖麻毒素作为膜转运的标志物,辣根过氧化物酶(HRP)作为液相转运的标志物,研究了莫能菌素对滤膜生长的犬肾Madin-Darby(MDCK)细胞的内吞作用、转胞吞作用、再循环以及向高尔基体转运的影响。莫能菌素(10微摩尔)刺激两种标志物在基底外侧向顶端方向的转胞吞作用增加约3倍。莫能菌素使HRP在相反方向(顶端到基底外侧)的转胞吞作用降低至对照的约50%,而蓖麻毒素的转胞吞作用略有增加。在存在莫能菌素的情况下,从顶端表面内吞的标志物的再循环减少,并且细胞内蓖麻毒素和HRP的积累均增加。蓖麻毒素向高尔基体的转运增加幅度与细胞内积累的增加幅度相同。当标志物从基底外侧添加时,未观察到莫能菌素对再循环或积累的影响,但蓖麻毒素向高尔基体的转运增加了近3倍。我们的结果表明,在不存在低内涵体pH值的情况下,基底外侧到顶端的转胞吞作用增加,并且表明膜结合标志物(蓖麻毒素)从顶端到基底外侧的转胞吞作用受莫能菌素的影响与液相标志物(HRP)不同。