Characterization of type A and type B CCK receptor binding sites in rat vagus nerve

Eric S. Corp, Jennifer McQuade, Timothy H. Moran, Gerard P. Smith

Research output: Contribution to journalArticle

Abstract

We employed quantitative receptor autoradiography to analyze pharmacological properties of 125I-Bolton Hunter cholecystokinin (CCK-8)-labeled binding sites in sections of rat cervical vagus nerve that had been ligated 24 h prior to extraction. Binding densities were detected in segments of nerve proximal and distal to the ligature. Analysis was confined to proximal segments. Saturation and competitive binding studies were carried out using sulphated CCK-8 and two selective CCK receptor antagonists: MK-329, to define type-A (CCKA) binding sites; and, L-365,260, to define type-B (CCKB) binding sites. Sulphated CCK-8 was the most potent inhibitor of vagal 125I-CCK binding (IC50 = 2 nM). Nonlinear curve fitting analysis of the CCK binding data favored the presence of a single class of vagal CCK receptors KDi = 1 nM). However, both MK-329 (IC50 = 18 nM) and L-365,260 (IC50 = 45 nM) completed for vagal 125I-CCK binding indicating the presence of CCKA and CCKB binding sites. Co-analysis of the antagonist binding data suggested that CCKA and CCKB receptors were transported in equal concentration within the vagus. MK-329 bound with high affinity to CCKA sites (Ki = 3 nM) and low-affinity to CCKB sites (Ki = 462 nM) while L-365,260 bound with high affinity to CCKB sites (Ki = 10 nM) and low-affinity to CCKA sites (Ki = 775 nM). These same ligands were used to characterize the specificity of 125I-CCK binding in the medial and lateral divisions of the nucleus of the solitary tract (NTS), two regions innervated by primary vagal afferents carrying CCK receptors. Sulphated CCK-8 was the most potent inhibitor of 125I-CCK binding in both regions of the NTS (IC50 = 0.6 nM, medial; IC50 = 0.4 nM, lateral). In the medial NTS, MK-329 was a potent inhibitor (IC50 = 21.8 nM) while MK-329 was a weak inhibitor (IC50 = 341 nM) of 125I-CCK binding. In contrast, in the lateral NTS, L-365,260 was a potent inhibitor (IC50 = 21.8 nM) while MK-329 was a weak inhibitor (IC> 1,0000 nM) of 125I-CCK binding. These results are consistent with the existence of two populations of vagal afferent fibers projecting to different regions of the NTS. One, containing CCKA receptors, projects principally to the medial NTS, while the other containing CCKB receptors, projects principally to the lateral NTS.

Original languageEnglish (US)
Pages (from-to)161-166
Number of pages6
JournalBrain research
Volume623
Issue number1
DOIs
StatePublished - Sep 24 1993

Keywords

  • Area postrema
  • Cholecystokinin receptor antagonist
  • Feeding
  • L-365,260
  • MK-329
  • Nucleus of the solitary tract
  • Receptor autoradiography
  • Visceral afferent

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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