Calcium Flux Assay

HTS223C - ChemiScreen™ D4 Dopamine Receptor Stable Cell Line

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HTS223C
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    The neurotransmitter dopamine is involved in a wide variety of neuroendocrine, emotional, cognitive and locomotor functions.  These activities of dopamine are mediated by a group of 5 G protein coupled receptors, 2 of which (D1 and D5) couple to Gs and 3 of which (D2, D3, D4) couple to Gi (Missale et al., 1998).  The D4 receptor is highly polymorphic, particularly in the third cytoplasmic loop, which contains from 2 to 7 repeats of 16 amino acids.  Individuals with 7 repeats have increased risk of developing ADHD (Thapar et al., 2005).  Mice engineered to lack D4 have reduced locomotor activity, increased sensitivity to drugs of abuse that elevate dopamine (cocaine, methamphetamine and ethanol), and reduced response to novelty (Rubinstein et al., 1997; Dulawa et al., 1999). Cloned human D4-expressing cell line is made in the Chem-5 host, which supports high levels of recombinant D4 expression on the cell surface and contains optimized levels of a recombinant promiscuous G protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists, antagonists and modulators at D4.

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    Item Unit of Measure: PK
    Contents: 2 vials of mycoplasma-free cells, 1 ml per vial.
    Storage: Vials are to be stored in liquid N2.
    Applications: Calcium Flux Assay
    Host Cell: Chem-5, an adherent rat hematopoietic cell line expressing endogenous G·15 protein as well as an exogenous proprietary promiscuous Gα protein.
    Exogenous Gene Expression: DRD4.4 cDNA (Accession Number: NM_000797; see CODING SEQUENCE below) expressed from a proprietary pHS plasmid.
    GMO: This product contains genetically modified organisms.
    Reference 1: 1. Chemel BR et al. (2006) WAY-100635 is a potent dopamine D4 receptor agonist. Psychopharmacology 188: 244-251.
    Reference 2: 2. Dulawa SC et al. (1999) Dopamine D4 receptor-knock-out mice exhibit reduced exploration of novel stimuli. J. Neurosci. 19: 9550-9556.
    Reference 3: 3. Millan MJ et al. (1998) S 18126 ([2-[4-(2,3-dihydrobenzo[1,4]dioxin-6-yl)piperazin-1-yl methyl]indan-2-yl]), a potent, selective and competitive antagonist at dopamine D4 receptors: an in vitro and in vivo comparison with L 745,870 (3-(4-[4-chloro
    Reference 4: 4. Missale C et al. (1998) Dopamine receptors: From structure to function. Physiol. Rev. 78:
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