Characterization of cholecystokinin receptors on the human gallbladder.
BACKGROUND
Several studies examined in vivo and in vitro biologic activity of the human gallbladder in response to cholecystokinin (CCK). However, few studies have demonstrated directly the interaction of CCK with receptors on the human gallbladder, which is responsible for this biologic activity.


METHODS
To characterize CCK receptors on human gallbladder tissue, gallbladders were removed from human donor grafts that were being used for liver transplantation. The gallbladders were rapidly frozen and sectioned for measurement of binding of 125I-Bolton-Hunter-labeled-CCK-8 and were cut into strips for in vitro bioassay.


RESULTS
Binding of 125I-BH-CCK-8 to human gallbladder was saturable, specific, and dependent on time, pH, and temperature. The binding was inhibited only by cholecystokinin-related peptides including CCK-8 (IC50 10 +/- 1.0 nmol/L) (mean +/- SD), des(SO3) CCK-8 (IC50 0.9 +/- 0.2 mumol/L), and gastrin-17-I (IC50 9.0 +/- 2.0 mumol/L) or specific CCK receptor antagonist L-364,718. Computer analysis of binding of 125I-BH-CCK-8 to gallbladder tissue showed a single class of binding sites with high affinity for CCK-8. Autoradiography localized binding of 125I-BH-CCK-8 only to the smooth muscle layer of the gallbladder. In the bioassay des(SO3) CCK-8 (EC50 1.2 +/- 0.7 mumol/L) and gastrin-17-I (EC50 4.5 +/- 2.4 mumol/L) were 150- and 563-fold less potent than CCK-8 (EC50 8.0 +/- 2.2 nmol/L). The relative potencies of CCK agonists for inhibiting binding of 125I-BH-CCK-8 agreed closely with their relative potencies for causing gallbladder contraction. The dose-response curve for CCK-8 alone to induce gallbladder contraction was not significantly different from those caused by CCK-8 plus 1 mumol/L tetrodotoxin or 1 mumol/L atropine.


CONCLUSIONS
These results characterized the CCK receptors on smooth muscle of human gallbladder as sulfate dependent and causing gallbladder contraction.
Tokunaga
Y
Y
California Pacific Medical Center, San Francisco 94115.
Cox
K L
KL
Coleman
R
R
Concepcion
W
W
Nakazato
P
P
Esquivel
C O
CO
D001345:Autoradiography; D001570:Benzodiazepinones / Q000494:pharmacology; D001665:Binding Sites; D002217:Carbachol / Q000494:pharmacology; D020109:Devazepide; D004305:Dose-Response Relationship, Drug; D005704:Gallbladder / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D005755:Gastrins / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes / Q000378:metabolism; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism*; D011949:Receptors, Cholecystokinin / Q000187:drug effects / Q000378:metabolism*; D012016:Reference Values; D012633:Secretin / Q000494:pharmacology; D012701:Serotonin / Q000494:pharmacology; D012844:Sincalide / Q000031:analogs & derivatives / Q000494:pharmacology; D013373:Substance P / Q000494:pharmacology; D013696:Temperature; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
D001570:Benzodiazepinones; D005755:Gastrins; D006728:Hormones; D007457:Iodine Radioisotopes; D011949:Receptors, Cholecystokinin; D012633:Secretin; C079530:desulfated sincalide; D012701:Serotonin; D013373:Substance P; D014660:Vasoactive Intestinal Peptide; C013550:gastrin 17; D002217:Carbachol; D020109:Devazepide; D012844:Sincalide