Polarized expression of the GFP-tagged rat V(1a) vasopressin receptor.
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.
Campos
D M
DM
Department of Physiology, Universidad Austral de Chile, Valdivia, Chile.
Reyes
C E
CE
Sarmiento
J
J
Navarro
J
J
González
C B
CB
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D011485:Protein Binding; D051381:Rats; D017483:Receptors, Vasopressin / Q000096:biosynthesis* / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection
D018076:DNA, Complementary; D008164:Luminescent Proteins; D017483:Receptors, Vasopressin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002118:Calcium