Posner B I, Verma A K, Patel B A, Bergeron J J
J Cell Biol. 1982 Jun;93(3):560-7. doi: 10.1083/jcb.93.3.560.
In previous studies we have shown that 125I-labeled prolactin is taken up by a receptor-dependent process and concentrated in an intact form in Golgi elements from female rat liver (J. Biol. Chem., 1979, 254:209-214). In this study we have examined the effect of colchicine on this uptake process into Golgi elements. Colchicine [25 mumol (10 mg)/100 gm body wt] was injected intraperitoneally in adult female rats, and hepatic Golgi fractions were prepared at 1, 2, and 3 h postinjection. The enzyme recoveries and morphological appearance of fractions from colchicine-treated and control (alcohol alone) animals were similar. At times greater than 1 h after colchicine there was a marked (greater than 60%) inhibition of uptake of 125I-ovine prolactin (125I-oPRL) into Golgi light and intermediate fractions but no inhibition of uptake into Golgi heavy and plasmalemma elements. At times from 2 to 45 min postinjection, 125I-oPRL was extracted from Golgi elements and found to be largely intact as judged by rebinding to receptors. The inhibitory effect of colchicine was seen at doses ranging from 0.25 mumol to 25 mumol/100 g body wt. Vincristine also inhibited 125I-oPRL uptake into the Golgi light and intermediate fractions but lumicolchicine had no inhibitory effect. There was a smaller effect of colchicine both at early (1 h) and later (3 h) times on the extent and pattern of 125I-insulin uptake. Colchicine treatment did not produce a significant change in lactogen receptor levels in the Golgi fractions. These results demonstrate that colchicine treatment inhibited the transfer of prolactin into Golgi vesicular elements. The much smaller effect on insulin uptake suggests that there may be differences in the manner in which the two hormones are handled in the course of internalization.
在先前的研究中我们已经表明,125I标记的催乳素通过受体依赖性过程被摄取,并以完整形式在雌性大鼠肝脏的高尔基体中浓缩(《生物化学杂志》,1979年,254:209 - 214)。在本研究中,我们检测了秋水仙碱对这种摄取进入高尔基体过程的影响。将秋水仙碱[25 μmol(10 mg)/100 g体重]腹腔注射到成年雌性大鼠体内,并在注射后1、2和3小时制备肝脏高尔基体组分。秋水仙碱处理组和对照组(仅注射酒精)动物的组分的酶回收率和形态外观相似。秋水仙碱处理后1小时以上,125I - 羊催乳素(125I - oPRL)摄取到高尔基体轻组分和中间组分中受到显著抑制(超过60%),但摄取到高尔基体重组分和质膜组分中未受抑制。在注射后2至45分钟期间,125I - oPRL从高尔基体组分中被提取出来,并且通过与受体重新结合判断发现其大部分保持完整。秋水仙碱在0.25 μmol至25 μmol/100 g体重的剂量范围内均可见抑制作用。长春新碱也抑制125I - oPRL摄取到高尔基体轻组分和中间组分中,但光秋水仙碱没有抑制作用。秋水仙碱在早期(1小时)和晚期(3小时)对125I - 胰岛素摄取的程度和模式的影响较小。秋水仙碱处理并未使高尔基体组分中催乳素受体水平发生显著变化。这些结果表明,秋水仙碱处理抑制了催乳素向高尔基体囊泡组分的转运。对胰岛素摄取的影响小得多,这表明两种激素在内化过程中的处理方式可能存在差异。