Autoradiographic localization of [125I-Tyr8]-bradykinin receptor binding sites in the guinea pig spinal cord

Abstract

The present study aimed to localize and characterize [125I-Tyr8]-BK binding sites in all major segments of the guinea pig spinal cord using in vitro quantitative receptor autoradiography. [125I-Tyr8]-BK specific binding sites were localized predominantly in superficial layers of the dorsal horn, with lamina II depicting the highest labelling. The density of specific binding in laminae I and III was moderate, whereas in other areas, i.e., laminae IV-X, lower amounts of labelling were noticed. The B2 receptor antagonists D-Arg[Hyp3,Thi5,D-Tic7,Oic8]-BK (Hoe 140), D-Arg[Hyp3,D-Phe7,Leu8]-BK, Tyr0,D-Arg[Hyp3,D-Phe7,Leu8]-BK, D-Arg[Tyr3,D-Phe7,Leu8]-BK, D-Arg[Hyp2,Thi5,8,D-Phe7]-BK, D-Arg[Hyp3,Leu8]-BK and D-Arg[Hyp3,Gly6,Leu8]-BK as well as unlabelled [Tyr8]-BK inhibited [125I-Tyr8]-BK binding with respective Ki values of 0.04, 12.4, 23.4, 34.5, 43.5, 33.5, 23.0, and 0.6 nM while B1 related molecules (Tyr0,des-Arg10-kallidin and [Leu8]-des-Arg9-BK) did not significantly inhibit [125I-Tyr8]-BK binding up to micromolar concentrations. These results indicate that the specific [125I-Tyr8]-BK binding sites present in the guinea pig spinal cord belong to the B2 receptor subtype. The high density of B2 binding sites in the substantia gelatinosa provides an anatomical evidence in favour of a role for BK as a modulator of nociceptive information.