Tada J, Sawa T, Yamanaka N, Shono M, Akamatsu T, Tsumura K, Parvin M N, Kanamori N, Hosoi K
Department of Physiology and Oral Physiology, Tokushima University School of Dentistry, 3-Kuramotocho, Tokushima-shi, Tokushima, 770-8504, Japan.
Biochem Biophys Res Commun. 1999 Dec 20;266(2):443-7. doi: 10.1006/bbrc.1999.1828.
A cDNA of rat aquaporin 5 (AQP5) was used to transfect to HSG (human salivary gland cells), and the trafficking mechanism was studied in vitro by confocal laser microscopy. The trafficking of AQP5 to the plasma membrane was induced by stimulation of AQP5-gene-transfected human salivary gland cells (HSGAQP5 cells) with thapsigargin, an inhibitor of endoplasmic Ca(2+)-ATPase, and or with A-23187, a calcium ionophore. Pretreatment of these cells with colchicine or vinblastine, microtubule inhibitors, prevented the trafficking induced by thapsigargin or A-23187. The trafficking event was not completely inhibited by cytochalasin B, a microfilament inhibitor. These results demonstrate that the trafficking of AQP5 vesicles to the plasma membrane is triggered by an increase in intracellular Ca(2+) and that the interaction of AQP5-containing vesicles with the cytoskeleton is involved in this trafficking.
用大鼠水通道蛋白5(AQP5)的互补DNA(cDNA)转染人唾液腺细胞(HSG),并通过共聚焦激光显微镜在体外研究其转运机制。用内质网Ca(2+) -ATP酶抑制剂毒胡萝卜素和/或钙离子载体A-23187刺激转染了AQP5基因的人唾液腺细胞(HSGAQP5细胞),可诱导AQP5向质膜的转运。用微管抑制剂秋水仙碱或长春花碱预处理这些细胞,可阻止毒胡萝卜素或A-23187诱导的转运。微丝抑制剂细胞松弛素B不能完全抑制转运事件。这些结果表明,细胞内Ca(2+)增加触发了含AQP5的囊泡向质膜的转运,且含AQP5的囊泡与细胞骨架的相互作用参与了这一转运过程。