title: Investigation of cholecystokinin receptors in the human lower esophageal sphincter.
abstract: OBJECTIVE
  To compare the binding of cholecystokinin (CCK)-8 to CCK receptors in sling and clasp fibers of the human lower esophageal sphincter.
  
  
  METHODS
  Esophageal sling and clasp fibers were isolated from eight esophagectomy specimens, resected for squamous cell carcinoma in the upper two thirds of the esophagus, which had been maintained in oxygenated Kreb's solution. Western blot was used to measure CCK-A and CCK-B receptor subtypes in the two muscles. A radioligand binding assay was used to determine the binding parameters of (3)H-CCK-8S to the CCK receptor subtypes. The specificity of binding was determined by the addition of proglumide, which blocks the binding of CCK to both receptor subtypes.
  
  
  RESULTS
  There was no significant difference between the sling and clasp fibers of the human lower esophageal sphincter in the amount of CCK-A [integrated optical density (IOD) value: 22.65 ± 0.642 vs 22.328 ± 1.042, P = 0.806] or CCK-B receptor protein (IOD value: 13.20 ± 0.423 vs 12.45 ± 0.294, P = 0.224) as measured by Western blot. The maximum binding of radio-labeled CCK-8S was higher in the sling fibers than in the clasp fibers (595.75 ± 3.231 cpm vs 500.000 ± 10.087 cpm, P < 0.001) and dissociation constant was lower (K(d): 1.437 ± 0.024 nmol/L vs 1.671 ± 0.024 nmol/L, P < 0.001). The IC₅₀ of the receptor specific antagonists were lower for the CCK-A receptors than for the CCK-B (P < 0.01).
  
  
  CONCLUSIONS
  CCK binding modulates the contractile function of the lower esophageal sphincter through differential binding to the CCK-A receptor on the sling and clasp fibers.
authors.affiliation: Jun-Feng Liu, Jian Zhang, Xin-Bo Liu, Department of Thoracic Surgery, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
authors.forename: Jun-Feng
authors.initials: JF
authors.lastname: Liu
authors.affiliation: Jun-Feng Liu, Jian Zhang, Xin-Bo Liu, Department of Thoracic Surgery, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Jun-Feng Liu, Jian Zhang, Xin-Bo Liu, Department of Thoracic Surgery, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
authors.forename: Xin-Bo
authors.initials: XB
authors.lastname: Liu
authors.affiliation: Jun-Feng Liu, Jian Zhang, Xin-Bo Liu, Department of Thoracic Surgery, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Drew
mesh_terms: D000368:Aged; D015153:Blotting, Western; D049630:Esophageal Sphincter, Lower / Q000473:pathology*; D016629:Esophagectomy; D004943:Esophagogastric Junction / Q000378:metabolism / Q000473:pathology*; D004947:Esophagus / Q000473:pathology; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D020128:Inhibitory Concentration 50; D007552:Isotonic Solutions; D008297:Male; D008875:Middle Aged; D011377:Proglumide / Q000737:chemistry; D043886:Receptor, Cholecystokinin A / Q000378:metabolism*; D043887:Receptor, Cholecystokinin B / Q000378:metabolism*; D012844:Sincalide / Q000378:metabolism*
chemical_list: D007552:Isotonic Solutions; C051865:Krebs-Ringer solution; D043886:Receptor, Cholecystokinin A; D043887:Receptor, Cholecystokinin B; D011377:Proglumide; D012844:Sincalide
keywords: Cholecystokinins; Cholecystokinins-A receptor; Cholecystokinins-B receptor; Clasp fibers; Lower esophageal sphincter; Radioligand binding; Sling fibers