MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT V2 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human V2 cDNA (GenBank Accession Number NM_000054.2) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM/F12, 10% FBS, 10 µg/mL puromycin
Background: V2, a G protein coupled receptor, also called arginine vasopressin receptor 2 (AVPR2), is a receptor for neurohypophyseal peptide [Arg8]-vasopressin. It is expressed in the kidney tubule, mainly in the membrane of cells of the distal convoluted tubule and collecting ducts, in fetal lung tissue and lung cancer. In the kidney, V2 receptor responds to vasopressin and activates mechanisms that concentrate urine and maintain water homeostasis in the body. Loss of function of the V2 receptor results in nephrogenic diabetes insipidus. The primary transduction mechanism is via adenylate cyclase stimulation to raise intracellular cAMP levels. Selective antagonists for V2 can be used to treat hyponatremia.
Application: Functional assays
Figure 1. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. a) Dose-dependent increase of intracellular cAMP upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). b) Dose-dependent inhibition of cAMP upon treatment with antagonist. Figure 3. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
van den Ouweland AM, Dreesen JC, Verdijk M, Knoers NV, Monnens LA, Rocchi M, van Oost BA (1992). “Mutations in the vasopressin type 2 receptor gene (AVPR2) associated with nephrogenic diabetes insipidus”. Nature Genetics 2(2):99-102.
Zmily HD, Daifallah S, Ghali JK (2011). “Tolvaptan, hyponatremia, and heart failure”. International Journal of Nephrology and Renovascular Disease 4:57-71.