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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. The endogenous catecholamines, epinephrine and norepinephrine have profound effects on smooth muscle activity, cardiac function, carbohydrate and fat metabolism, hormone secretion, neurotransmitter release, and central nervous system actions. These activities are mediated by GPCRs belonging to two subfamilies, the ?- and ?-adrenoceptors (Bylund et al., 1994). The three members of the alpha1 subclass of adrenoceptors, alpha1A, alpha1B and alpha1D, couple to Gq, and promote contraction of vascular and urinary tract smooth muscle, relaxation of intestinal smooth muscle, increased contractile force in the heart, and glycogenolysis and gluconeogenesis in the liver. The different subtypes have overlapping distributions and variably contribute to these effects depending on species and tissue, the alpha1A subtype plays a prominent role in urogenital smooth muscle contraction and renal artery contraction (Hrometz et al., 1999, Ruffolo and Hieble, 1999). Activation of alpha1 adrenoceptors also influences cell proliferation, alpha1A inhibits cell growth by arresting progression at the G1/S transition (Shibata et al., 2003). The alpha1A subtype undergoes alternative splicing to generate four variants that differ at their C-termini, although these variants appear to be functionally identical (Chang et al., 1998). Millipore's cloned human alpha1A -expressing cell line is made in the Chem-1 host, which supports high levels of recombinant alpha1A expression on the cell surface for functional detection via the calcium signaling pathway. Thus, the cell line is an ideal tool for screening agonists, antagonists and modulators at alpha1A adrenergic receptors.