Open AccessCloning of ligand-specific cell lines via gene transfer: identification of a D2 dopamine receptor subtype.
Author(s) -
Richard D. Todd,
Tejvir S. Khurana,
P. Sajovic,
Kevin Stone,
Karen L. O’Malley
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.24.10134
Subject(s) - spiperone , microbiology and biotechnology , expression cloning , transfection , cloning (programming) , gene , cell culture , receptor , intracellular , biology , flow cytometry , dopamine receptor , chemistry , genetics , complementary dna , computer science , programming language
Using rat genomic DNA, we have established a transfected mouse fibroblast cell line that expresses a spiperone binding site with the pharmacological characteristics of a D2 dopamine receptor. The expressed D2 receptors are the product of a gene that is distinct from that reported by Bunzow et al. [Bunzow, J. R., Van Tol, H. H. M., Granoly, D. K., Albert, P., Salon, J., Christie, M., Machida, C. A., Neve, K. A. & Civelli, O. (1988) Nature (London) 336, 783-787]. Flow cytometry with the Ca2+-sensitive dye indo-1 demonstrated that activation of the expressed D2 sites resulted in increases in intracellular calcium that were dependent on the influx of external Ca2+. These general cloning procedures should be applicable to the production of cell lines expressing a variety of genes for which only functional assays are available.