title: Characterization of [125I]S(-)5-OH-PIPAT binding to dopamine D2-like receptors expressed in cell lines.
abstract: Recently, [125I]S(-)5-OH-PIPAT [5-hydroxy-2-(N-n-propyl-N-3'-iodo-2'-propenyl)aminotetralin], a derivative of S(-)5-OH-DPAT (5-hydroxy-N,N-di-n-propyl-aminotetralin), was reported to be a better radioiodinated dopamine D2-like receptor ligand than the previously reported iodinated ligand, [125I]R(+)7-OH-PIPAT. Therefore, in the present study, the binding profile of [125I]S(-)5-OH-PIPAT to D2-like receptors expressed in cell lines was established. High binding affinity (Kd = 0.3-0.4 nM) and NaCl sensitivity were displayed with this ligand in membranes of human embryonic kidney (HEK293) cells expressing either human D2 or rat D3 receptors and in Chinese hamster ovary (CHO) cells expressing human dopamine D4 receptors. Specific binding to D2 and D4 receptors was significantly increased in the presence of 2 mM MgCl2 and decreased in the presence of 100 microM 5'-guanylyl-imidodiphosphate (GMP-PNP). This finding is consistent with reports that 2-aminotetralin compounds display agonist properties. The specific binding to D3 receptors however, was not affected by either MgCl2 or GMP-PNP. This lack of GMP-PNP sensitivity for D3 receptors may result from inadequate G protein-receptor coupling in this cell line. The rank order of potency for inhibition of [125I]S(-)5-OH-PIPAT binding with various dopamine agents was consistent with reported values for D2, D3 and D4 receptors. In membranes prepared from Spodoptera frugiperda (Sf9) cells infected with baculovirus that contains DNA encoding D3 receptors, [125I]S(-)5-OH-PIPAT recognized only 70% of the receptor population labeled by [125I]NCQ298. This new ligand offers several unique advantages, including high specific activity, high binding affinity and selectivity for D2-like receptors, that make it an excellent probe for the investigation and the characterization of dopamine D2-like receptors.
authors.affiliation: Department of Pharmacology, University of Pennsylvania, Philadelphia 19104, USA. jvess@sunmac.spect.upenn.edu
authors.forename: J M
authors.initials: JM
authors.lastname: Vessotskie
authors.forename: M P
authors.initials: MP
authors.lastname: Kung
authors.forename: S
authors.initials: S
authors.lastname: Chumpradit
authors.forename: H F
authors.initials: HF
authors.lastname: Kung
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D018491:Dopamine Agonists / Q000494:pharmacology*; D018492:Dopamine Antagonists / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007457:Iodine Radioisotopes; D008024:Ligands; D011869:Radioligand Assay; D017448:Receptors, Dopamine D2 / Q000187:drug effects*
chemical_list: D018491:Dopamine Agonists; D018492:Dopamine Antagonists; D007457:Iodine Radioisotopes; D008024:Ligands; D017448:Receptors, Dopamine D2