ChemiSCREEN™ Rat Recombinant V1_a Vasopressin Receptor Calcium-Optimized Stable Cell Line

ChemiSCREEN™ Rat Recombinant V1_a Vasopressin Receptor Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length rat V_1a cDNA

Arginine vasopressin (AVP) is a 9 amino acid peptide that functions as an antidiuretic, vasoconstrictor and neurotransmitter. The three vasopressin receptors, V_1A, V_1B and V₂, are GPCRs; V_1A and V_1B couple to Gq and calcium release, whereas V₂ couples to Gs. V1 receptors regulate fluid homeostasis by stimulating contraction of glomerular mesangial cells and renal vascular smooth muscle cells (Birnbaumer, 2000). Chemicon's cloned rat V_1A-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant V_1A expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to enhance coupling of the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between V_1A and its ligands.

Calcium flux assay, ligand binding assays

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The protein encoded by this gene acts as receptor for arginine vasopressin. This receptor belongs to the subfamily of G-protein coupled receptors which includes AVPR1B, V2R and OXT receptors. Its activity is mediated by G proteins which stimulate a phosphatidylinositol-calcium second messenger system. The receptor mediates cell contraction and proliferation, platelet aggregation, release of coagulation factor and glycogenolysis.

FUNCTION: SwissProt: P37288 # Receptor for arginine vasopressin. The activity of this receptor is mediated by G proteins which activate a phosphatidyl- inositol-calcium second messenger system. Has been involved in social behaviors, including affiliation and attachment.
SIZE: 418 amino acids; 46800 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P37288 ## Belongs to the G-protein coupled receptor 1 family.
MISCELLANEOUS: Differences in regional receptor expression in the brain as well as differences in social behavior may result from a highly variable repetitive sequence in the 5' flanking region of AVPR1A. One such allelic variant has been linked to autism.

Rat

EC50 for calcium mobilization by vasopressin: 0.583 nM.

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10⁶ cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

V1A, rat

P37288

peptide

Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO₂. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca⁺⁺ and Mg⁺⁺-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.

Vasopressin

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A

2 x 10⁶ cells/vial

• AVPR1A
• AVPR1
• V1aR

GPCRs

ChemiSCREEN™ Rat Recombinant V1_a Vasopressin Receptor Calcium-Optimized Stable Cell Line

NM_000706.3

DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin