Campos D M, Reyes C E, Sarmiento J, Navarro J, González C B
Department of Physiology, Universidad Austral de Chile, Valdivia, Chile.
Biochem Biophys Res Commun. 2001 Nov 30;289(2):325-8. doi: 10.1006/bbrc.2001.5981.
We investigated the targeting of the V(1a) receptor fused with the green fluorescence protein (V(1a)R-GFP) in polarized MDCK cells. Cells expressing V(1a)R-GFP displayed binding to vasopressin (AVP) and AVP-induced calcium responses, similar to cells expressing the wild-type V1a receptor. Interestingly, as with the wild-type V(1a)R, V(1a)R-GFP is preferentially distributed in the basolateral side of MDCK cells as monitored by confocal microscopy. Furthermore, AVP induced internalization of GFP-tagged receptors. Therefore, the GFP-tagged V(1a) receptor retains all the sorting signals of the wild-type receptor and offers an excellent system to elucidate the mechanisms of cell trafficking of V(1a) receptors.
我们研究了与绿色荧光蛋白融合的V(1a)受体(V(1a)R-GFP)在极化的MDCK细胞中的靶向作用。表达V(1a)R-GFP的细胞表现出与加压素(AVP)的结合以及AVP诱导的钙反应,这与表达野生型V1a受体的细胞相似。有趣的是,通过共聚焦显微镜监测发现,与野生型V(1a)R一样,V(1a)R-GFP优先分布在MDCK细胞的基底外侧。此外,AVP诱导了GFP标记受体的内化。因此,GFP标记的V(1a)受体保留了野生型受体的所有分选信号,并为阐明V(1a)受体的细胞转运机制提供了一个极好的系统。