Differential modulation by GTPγS of agonist and inverse agonist binding to h5‐HT 1A receptors revealed by [ 3 H]‐WAY100,635

The interaction of serotonergic ligands at human (h) 5‐HT 1A receptors expressed in Chinese hamster ovary cells was examined with the selective ‘neutral’ 5‐HT 1A antagonist [ 3 H]‐WAY100,635. Its binding was saturable ( K D =0.056 n M ) with a B max (3.65 pmol mg −1 ) significantly higher than that of two other selective 5‐HT 1A radioligands: the partial agonist, [ 3 H]‐S15535 (2.77 pmol mg −1 ) and the agonist, [ 3 H]‐8‐OH‐DPAT (2.02 pmol mg −1 ). The influence of GTPγS (100 μ M ) on the binding affinity of 15 serotonergic agonists, partial agonists, antagonists and inverse agonists was investigated in competition binding experiments with [ 3 H]‐WAY100,635. Agonists, including 5‐HT, 8‐OH‐DPAT and buspirone, displayed biphasic isotherms which shifted to the right in the presence of GTPγS. In contrast, isotherms of the inverse agonists, methiothepin, (+)butaclamol and spiperone, were shifted to the left in the presence of GTPγS. Unlabelled WAY100,635 was the only ligand that was unaffected by GTPγS, consistent with ‘neutral’ antagonist properties. The magnitude of affinity changes induced by GTPγS for 13 ligands was highly correlated ( r =0.98) with their efficacy (positive and negative) previously determined by [ 35 S]GTPγS binding. In contrast, the napthylpiperazine derivative and high efficacy agonist, S14506, displayed only a modest GTPγS shift, in accordance with previous indications of ‘atypical’ binding properties of this ligand. A further full agonist, S14671, which is chemically closely‐related to S14506, also displayed a minimal GTPγS shift, underpinning this observation. In conclusion, [ 3 H]‐WAY100,635 constitutes a useful neutral antagonist radioligand for the characterization of drug actions at h5‐HT 1A receptors. GTPγS‐induced affinity changes of agonist and inverse agonist competition isotherms generally correlate well with ligand efficacy, with the notable exception of two chemically‐similar agents, S14506 and S14671, which are efficacious agonists, yet relatively insensitive to h5‐HT 1A receptor/G‐protein coupling changes.British Journal of Pharmacology (2001) 132 , 518–524; doi: 10.1038/sj.bjp.0703832