Detection and characterization of a 5HT1D serotonin receptor-GTP binding protein interaction in porcine and bovine brain.

Radioligand binding studies were performed to characterize serotonin 5HT1D binding sites in porcine and bovine brain. 3H-5HT binding, in the presence of 1 microM (+/-)pindolol (to block 5HT1A and 5HT1B receptors) and 100 nM mesulergine (to block 5HT1C receptors), was specific, saturable, and of high affinity. In porcine and bovine cortex and striatum the majority of 5HT1 sites (80%-90%) were of the 5HT1D subtype. In competition experiments 8-OH-DPAT, TFMPP, mesulergine, DOB, and ICS 205-930 had low affinity for 3H-5HT-labeled 5HT1D sites, indicating that the pharmacology of the 5HT1D site is distinct from previously identified 5HT1A, 5HT1B, 5HT1C, 5HT2, and 5HT3 sites. Guanyl nucleotides, GTPgammaS, and Gpp(NH)p, and divalent cations potently modulated the binding of 3H-5HT to 5HT1D sites in porcine and bovine striatum. Mg++ ions increased the number and affinity of 3H-5HT-labeled 5HT1D sites, while guanyl nucleotides decreased the number of 3H-5HT-labeled 5HT1D sites. These results demonstrate the presence of the 5HT1D receptors in porcine striatum and bovine cortex and provide direct demonstration that the radioligand binding assay for the 5HT1D receptor can monitor the interaction of this receptor with a GTP-binding protein.