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Beschreibung

Millipore's Ready-to-Assay GPCR frozen cells are designed for simple, rapid calcium assays with no requirement for intensive cell culturing. Millipore has optimized the freezing conditions to provide cells with high viability and functionality post-thaw. The user simply thaws the cells and resuspends them in media, dispenses cell suspension into assay plates and, following over night recovery, assays for calcium response. Arginine vasopressin (AVP) is a 9 amino acid peptide that functions as an antidiuretic, vasoconstrictor and neurotransmitter. The three vasopressin receptors, V1A, V1B and V2, are GPCRs, V1A and V1B couple to Gq and calcium release, whereas V2 couples to Gs (Birnbaumer, 2000). The V1B receptor is expressed prominently in the anterior pituitary, where it mediates vasopressin-induced release of ACTH (Tanoue et al., 2004). A selective antagonist of V1B has recently been developed and shown to reduce depression, anxiety, and aggression in rodents (Blanchard et al, 2005, Griebel et al., 2002). Millipore's cloned human V1B-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant V1B expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between V1B and its ligands.