Cloning of ligand-specific cell lines via gene transfer: identification of a D2 dopamine receptor subtype.
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.
Todd
R D
RD
Department of Psychiatry, Washington University School of Medicine, Saint Louis, MO 63110.
Khurana
T S
TS
Sajovic
P
P
Stone
K R
KR
O'Malley
K L
KL
D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002999:Clone Cells; D004247:DNA / Q000235:genetics; D005796:Genes; D015698:Genomic Library; D007700:Kinetics; D007739:L Cells / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D051381:Rats; D011954:Receptors, Dopamine / Q000235:genetics* / Q000378:metabolism; D017448:Receptors, Dopamine D2; D015183:Restriction Mapping; D013134:Spiperone; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D015345:Oligonucleotide Probes; D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2; D013134:Spiperone; D004247:DNA; D013937:Thymidine Kinase; D002118:Calcium