CCK-induced Contraction Research Articles

Molecular cloning of cholecystokinin (CCK) and CCK-A receptor and mechanism of CCK-induced gastrointestinal motility in Suncus murinus

Cholecystokinin (CCK) is a peptide hormone mainly secreted by small intestinal endocrine I-cells and functions as a regulator of gallbladder contraction, gastric emptying, gastrointestinal (GI) motility, and satiety. The cellular effects of CCK in these peripheral tissues are predominantly mediated via CCK-A receptors which are found in smooth muscles, enteric neurons, and vagal afferent neurons in humans and animal models. Although various functions of CCK have been reported to be neurally mediated, it can also stimulate contraction via the CCK receptor on the smooth muscle. However, the entire underlying neural and cellular mechanisms involved in CCK-induced GI contractions are not clearly understood. Here, we first determined the cDNA and amino acid sequences of CCK and CCK-A receptor along with the distributions of cck mRNA and CCK-producing cells in house musk shrew (Suncus murinus, the laboratory strain named as suncus) and examined the mechanism of CCK-induced contraction in the GI tract. Mature suncus CCK-8 was identical to other mammalian species tested here, and suncus CCK-A receptor presented high nucleotide and amino acid homology with that of human, dog, mouse, and rat, respectively. Suncus CCK mRNA and CCK-producing cells were found mainly in small intestine and colon. In the organ bath study, CCK-8 induced dose-dependent contractions in the suncus stomach, duodenum, and jejunum, and these contractions were inhibited by atropine and CCK-A receptor antagonist. These results suggest that CCK-8-induced contraction is mediated in the myenteric cholinergic neural network and that CCK-A receptor is partly responsible for CCK-8-induced contractions. This study indicates that suncus is a useful animal model to study the functions of CCK involved in GI motility.

Mast Cell-Mediated Changes in Smooth Muscle Contractility during Mouse Giardiasis

Giardia intestinalis is a significant cause of diarrheal disease worldwide. Infections in animal models have been shown to cause changes in gastrointestinal transit that depend on adaptive immune responses and are mediated, in part, through neuronal nitric oxide synthase. Nitric oxide is an inhibitory neurotransmitter, and we therefore investigated potential excitatory pathways that might be involved in the response to Giardia infection. Infected mice exhibited increased spontaneous and cholecystokinin (CCK)-induced contractions of longitudinal smooth muscle. In contrast, enhanced contractile responses were not observed in response to acetylcholine, 5-hydroxytryptamine, or the protease-activated receptor-1 agonist peptide TFFLR. Giardia-induced changes in smooth muscle function appear to be mediated primarily by mast cells, as both spontaneous and CCK-induced contractions were blocked by pretreatment with either ketotifen or compound 48/80. Together, these data support a model in which CCK release triggers mast cell degranulation, leading to increases in smooth muscle contractility. These contractions, coupled with nitric oxide-mediated muscle relaxation, promote intestinal transit and parasite elimination.

Adaptation to low-protein diet increases inhibition of gastric emptying by CCK

Chronic nutritional disorders such as protein malnutrition are associated with delayed gastric emptying and increased postprandial cholecystokinin (CCK) levels. This study investigated the mechanisms involved in gastric emptying adaptation to low-protein diet. Two groups of 12 rats were adapted to a low-protein (LPD) or standard diet (SD) for 3 weeks. As compared to rats fed a SD, in rats adapted to a LPD gastric emptying was delayed, whereas postprandial CCK levels were increased. LPD enhanced antral muscle contractile response to CCK and cerulein without altering response to acetylcholine. This increased contractility was associated with up-regulation of CCK-A receptor mRNA levels in antral muscle. Our data suggest that modulation of gastric emptying after adaptation to a low-protein diet involves up-regulation of both CCK-A receptors and CCK-induced contraction of antral smooth muscle.

Contribution of different phospholipases and arachidonic acid metabolites in the response of gallbladder smooth muscle to cholecystokinin

Guinea pig gallbladder muscle strips were used to investigate the contribution of different sources of diacylglicerol (DAG) in the cholecystokinin (CCK)-induced contraction. The involvement of arachidonic acid (AA) in this response was also investigated. Three distinct pathways for DAG production were investigated with specific phospholipase (PL) inhibitors. U-73122 (10 μM) was used for inhibition of phosphoinositide-specific-PLC (PI-PLC), D-609 (100 μM) for phosphatidylcholine specific-PLC (PC-PLC), and propranolol (100 μM) for phospholipase D (PLD). Separate or combined inhibition of each of these enzymes showed that the CCK-induced output of DAG involves the parallel activation of each of these phospholipases. Thus, after inhibition of a PL subtype, the remaining subtypes were able to functionally compensate in mediating CCK-induced contraction. Inhibition of AA production via DAG-lipase or phospholipase A 2 (PLA 2) was accomplished using RHC-80267 (40 μM), mepacrine (100 μM) and 4-BPB (100 μM). These inhibitors diminished contractile response, indicating that AA is an important modulator of CCK-induced contraction. Indomethacin (10 μM) and nordihydroguaiaretic acid (NDGA, 100 μM), which inhibit subsequent steps in AA metabolism through the cyclooxygenase and 5-lipooxygenase pathways, also inhibited contractions. Taken together, these results show that CCK redundantly activates PC-PLC, PI-PLC and PLD, to produce DAG, which in turn stimulates PKC and provides a substrate for the generation of AA. sPLA 2 is also a source of AA, whose metabolites are, in part, responsible for determining the magnitude of the CCK-evoked contraction.

Effects of bile acids on the muscle functions of guinea pig gallbladder.

Hydrophobic bile acids impair gallbladder emptying in vivo and inhibit gallbladder muscle contraction in response to CCK-8 in vitro. This study was aimed at determining the mechanisms of muscle cell dysfunction caused by bile acids in guinea pig gallbladders. Muscle cells were obtained by enzymatic digestion. Taurochenodeoxycholic acid (TCDC), a hydrophobic bile acid, caused a contraction of up to 15% and blocked CCK-induced contraction. Indomethacin abolished the TCDC-induced contraction. Hydrophilic bile acid tauroursodeoxycholic acid (TUDC) had no effect on muscle contraction but prevented the TCDC-induced contraction and its inhibition on CCK-induced contraction. Pretreatment with NADPH oxidase inhibitor PH2I, xanthine oxidase inhibitor allopurinol, and free-radical scavenger catalase also prevented TCDC-induced contraction and its inhibition of the CCK-induced contraction. TCDC caused H2O2 production, lipid peroxidation, and increased PGE2 synthesis and activities of catalase and SOD. These changes were significantly inhibited by pretreatment of PH2I or allopurinol. Inhibitors of cytosolic phospholipase A2 (cPLA2), protein kinase C (PKC), and mitogen-activating protein kinase (MAPK) also blocked the TCDC-induced contraction. It is concluded that hydrophobic bile acids cause muscle cell dysfunction by stimulating the formation of H2O2 via activation of NADPH and xanthine oxidase. H2O2 causes lipid peroxidation and activates cPLA2 to increase PGE2 production, which, in turn, stimulates the synthesis of free-radical scavengers through the PKC-MAPK pathway.

Pharmacological characterization of cholecystokinin receptors mediating contraction of human gallbladder and ascending colon

Cholecystokinin (CCK) produces contractions of gallbladder and colon in a number of different species. Although the effects of CCK on the human gallbladder are relatively well documented, the CCK receptors in the human colon have not been clearly characterised. Therefore, in this study, the CCK receptors in the human gallbladder and colon were compared using pharmacological techniques. Contraction of specimens of the human tissue was measured using in vitro organ bath bioassay. The effect of selective concentrations of CCK 1 and CCK 2 receptor antagonists (L-364,718 and JB93182, respectively) was determined on agonist concentration–effect ( E/[ A]) curves obtained by cumulative dosing with sulphated CCK. The CCK 1 antagonist L-364,718 produced a rightward shift of the CCK-8S [ E/[ A] curve in the human gallbladder (p A 2=9.15±0.26) and ascending colon (p A 2=9.20±0.33). In both tissues, the CCK 2 receptor antagonist, JB93182, had no effect on the CCK E/[ A] curves. In addition, in the colon, pentagastrin responses were inhibited by L-364,718 but unaffected by JB93182. These data indicate that the CCK-induced contraction of the human colon and gallbladder smooth muscle is mediated solely through the CCK 1 receptor subtype, and the antagonist affinity estimates are consistent with those previously obtained in experiments on animal tissue.

Different affinity states of CCK 1 receptors on pancreatic acini and gastric smooth muscle in the rat

It has recently been shown that—after chronic cholecystokinin (CCK) treatment—an adaptation of pancreatic secretory but not gastric motor function does occur. Recent studies indicate that the CCK 1-receptor exists in two (i.e. high and low) affinity states, which could be distinguished by the CCK-analogue JMV-180. CCK occupancy of high and low affinity sites is thought to be related to the initiation of different intracellular events and consequent biological responses. Affinity states of CCK 1-receptors on pancreas and gastrointestinal (GI) smooth muscle could be different and this can offer an explanation for the different effects of CCK on pancreatic and gastric growth. We therefore studied the affinity states of CCK 1-receptors on isolated rat pancreatic acini and gastric smooth muscle preparations. When acini were incubated with increasing concentrations of CCK-8, a biphasic (i.e. stimulation followed by inhibition) effect on amylase release was observed. JMV-180 caused only stimulation of enzyme release and combined JMV-180 and CCK stimulation (at submaximal doses) resulted in an additive secretory response. CCK-8 induced contractions of pyloric, antral and fundic muscle in a concentration-dependent manner. The response was monophasic, reaching a plateau. JMV-180 had only a very weak effect on these preparations. On the contrary, it inhibited CCK-induced contractions in a competitive manner, the concentration–response curve to CCK being shifted to the right by the CCK analogue. Our data suggest that the affinity states of CCK 1-receptors on rat pancreatic and gastric tissue are different. On pancreatic acini CCK 1-receptors exist in both high- and low-affinity states whose occupation is followed by the sequence of intracellular events leading to growth. In contrast, occupation of low affinity receptors (the only ones present in the GI smooth muscle) does not lead to cell proliferation. This difference therefore explains the different adaptive response of the pancreas and the stomach to chronic CCK administration. Furthermore, different affinity states of CCK 1-receptors may mediate different functions of the digestive tract.

Signal transduction pathways mediating CCK-induced gallbladder muscle contraction.

The signal transduction that mediates CCK-induced contraction of gallbladder muscle was investigated in the cat. Contraction was measured by scanning micrometry in single muscle cells isolated enzymatically with collagenase. Production of D-myo-inositol 1,4, 5-trisphosphate (IP3) and sn-1,2-diacylglycerol (DAG) was quantitated using HPLC and TLC, respectively. Protein kinase C (PKC) activity was determined by measuring the phosphorylation of a specific substrate peptide from myelin basic protein, Ac-MBP-(4-14). CCK-induced contraction was blocked by incubation in strontium medium, pertussis toxin (PTx), and antibodies against Gialpha3 or betagamma-subunits but was not blocked by Ca2+-free medium or by antibodies against Gq/11alpha, Gialpha1-2, or Goalpha. The contraction induced by CCK was inhibited by the phospholipase C (PLC) inhibitor U-73122, anti-PLC-beta3 antibody, and the IP3 receptor antagonist heparin but was not inhibited by the the phospholipase D inhibitor propranolol or antibodies against PLC-beta1 or PLC-beta2. Western blot analysis of gallbladder muscle revealed the presence of PLC-beta2 and PLC-beta3 but not PLC-beta1. CCK caused a 94% increase in IP3 generation and an 86% increase in DAG generation. A low dose of CCK caused PKC translocation, and CCK-induced contraction was blocked by the PKC inhibitor H-7. A high dose of CCK, however, caused no PKC translocation, and its contraction was blocked by the calmodulin antagonist CGS9343B. In conclusion, CCK contracts cat gallbladder muscle by stimulating PTx-sensitive Gi 3 protein coupled with PLC-beta3, producing IP3 and DAG. Low doses activate PKC, whereas high doses activate calmodulin.

Signal transduction pathways associated with contraction during development of the feline gastric antrum

BACKGROUND & AIMS: Unlike adult antral cells, feline newborn antral cells are unable to contract in response to agonists in the absence of extracellular calcium or in response to exogenous inositol 1,4,5- triphosphate (IP3) after permeabilization. Changes in intracellular pathways that are associated with these differences were examined.METHODS: In adult and kitten antrum isolated smooth muscle cell contraction, levels of 1,2-diacylglycerol (DAG) and IP3 were assessed in response to cholecystokinin (CCK).RESULTS: CCK-induced contraction was transient in the adult and sustained in the kitten. U73122 blocked contraction in adult antral cells but not kitten antral cells. In adult antral tissue, CCK (10(-7) mol/L) caused an early transient increase in the level of DAG, whereas in the newborn antrum, CCK (10(-7) mol/L) caused a sustained increase in the DAG level for up to 4 minutes. IP3 showed an early increase in both age groups. Newborn contraction is associated with an initial increase in IP3 and sustained elevation of DAG levels, whereas in adult antral cells, there is a transient increase in both IP3 and DAG.CONCLUSIONS: The relative inaccessibility of intracellular calcium stores in the newborn is associated with age- related differences in signal transduction pathways.(Gastroenterology 1997 Aug;113(2):507-13)

Different pathways mediate cholecystokinin actions in cholelithiasis.

Smooth muscle from gallbladders with cholesterol stones exhibits impaired response to cholecystokinin (CCK). This study investigated whether the impaired response is mediated by different signal-transduction pathways responsible for CCK-induced contraction in prairie dog and human gallbladders with cholesterol stones. Gallbladder muscle cells were isolated enzymatically to study contraction. Protein kinase C (PKC) activity was measured by examining the phosphorylation of a specific substrate peptide from myelin basic protein Ac-MBP-(4-14). Gallbladder muscle cells from high-cholesterol-fed prairie dogs contracted less in response to CCK octapeptide (CCK-8) than those from the control group. However, inositol-1,4,5-trisphosphate (IP3), diacylglycerol, and guanosine 5'-O-(3-thiotriphosphate) induced the same magnitudes of contraction in these two groups. In control prairie dog and human gallbladders, the maximal contraction caused by 10(-8) M CCK-8 was blocked by the calmodulin antagonist CGS9343B but not by the PKC inhibitor H-7. Conversely, in gallbladders with cholesterol stones from prairie dogs or human patients, the maximal contraction induced by 10(-8) M CCK-8 was blocked by H-7 and chelerythrine but not by CGS9343B. In these gallbladders CCK-8 caused a significant PKC translocation from the cytosol to the membrane. High CCK concentrations may activate the calmodulin-dependent pathway in functionally normal gallbladder muscle and the PKC-dependent pathway in muscle from gallbladders with cholesterol stones. The defect of gallbladder muscle after cholesterol feeding and stones might reside in the steps before G protein activation.

Effect of modulating voltage-dependent calcium channels on cholecystokinin and acetylcholine-induced contractions of the guinea pig gallbladder

The aim of this study was to investigate the role of extracellular Ca 2+ utilization in cholecystokinin (CCK) and acetylcholine-induced guinea pig gallbladder contractions by using agents that modulate influx of extracellular Ca 2+ through voltage-dependent calcium channels. METHODS: Guinea pig gallbladder muscle strips were studied isometrically at L max in vitro. Results: (1) Acetylcholine and CCK caused dose-dependent contractions, with ED max of 10 −4 to 10 −6 M, respectively. (2) Preventing influx of extracellular Ca 2+ by incubation in Ca 2+-free/0.1 mM EGTA solution inhibited the acetylcholine (10 −4 M)-induced contraction by 60 ± 3% compared to only 46 ± 5% ( P $ ̌ 0.05 ) for CCK (10 −6 M)-induced contraction. (3) Nifedipine (3 μM) inhibited the response to acetylcholine (10 −4 M) by 54 ± 3%, compared to only 34 ± 3% ( P < 0.01) for CCK (10 −6 M). (4) Bay K 8644 (10 −7 M) significantly increased ( P < 0.05) the contractile responses to low doses of each agonist: acetylcholine (10 −6 M) by 121 ± 44% and CCK (10 −9 M) by 94 ± 31%, but had no effect on the contraction to the ED max of each agonist. Conclusions: These studies demonstrate: (1) acetylcholine and CCK cause guinea pig gallbladder contraction by both intracellular Ca 2+ release and influx of extracellular Ca 2+ through voltage-dependent calcium channels; (2) the CCK-induced contraction is more dependent on intracellular Ca 2+ than is acetylcholine; and (3) acetylcholine and CCK-induced contractions can by modulated by manipulating influx of extracellular Ca 2+ through voltage-dependent calcium channels.

Cholecystokinin-coupled intracellular signaling in human gallbladder muscle

Background/Aims: It has been shown that cholecystokinin (CCK) contracts the gallbladder muscle by utilizing intracellular calcium, but the intracellular pathways have not been elucidated. The present study was designed to characterize the signal transduction pathways that mediate CCK-induced contraction of human gallbladder muscle. Methods: Single muscle cells were isolated from human gallbladders by enzymatic digestion with collagenase. Permeable cells were obtained by incubation with saponin. Protein kinase C (PKC) activity was determined by measuring the phosphorylation of a specific substrate peptide from myelin basic protein, Ac-MBP(4–14). Results: The inositol-1,4,5-tris-phosphate (IP3) antagonist heparin blocked the contractions induced by CCK. The PKC inhibitor H-7 blocked the contractions caused by low, but not high, concentrations of CCK and IP3. In contrast, the calmodulin inhibitor CGS9343B blocked the contractions induced by high, but not low, doses of CCK and IP3. Furthermore, exogenously activated calmodulin blocked the PKC-mediated contraction induced by diacylglycerol. Direct measurements of PKC activity showed that low, but not high, CCK concentrations caused PKC translocation. Conclusions: CCK contracts the gallbladder muscle via IP3-mediated calcium release. CCK activates the PKC pathway at low concentrations, whereas it activates the calmodulin pathway at high concentrations, which in turn inhibits the activation of PKC.

Galanin induces opposite effects via different intracellular pathways in smooth muscle cells from dog colon

Smooth muscle cells isolated by enzymatic digestion were used to determine the direct effects of galanin on circular and longitudinal muscle layers from dog proximal colon and to investigate the intracellular pathways involved in these effects. Effects of galanin were compared to those observed with other contracting [cholecystokinin octapeptide (CCK8)] and relaxing [vasoactive intestinal peptide (VIP)] agents. In longitudinal cells, galanin and CCK8 induced a contraction that was maximal at 1 n M galanin and 1 n M CCK8 and was 23.9 ± 4.5% and 23.4 ± 3.4%, respectively, of the length of resting cells. Incubation of cells in Ca 2+ -free medium or in the presence of nifedipine caused an inhibition of galanin-induced contraction whereas it had no effect on the contraction induced by CCK8. Vasoactive intestinal peptide, forskolin, and 8 bromo cAMP inhibited CCK-induced contraction but failed to inhibit contraction induced by galanin. The contraction induced by galanin was abolished; the CCK-induced contraction was unchanged by pertussis toxin. In circular cells, CCK8 induced a contraction that was maximal at 10 n M and was 24.2 ± 2.6%. Galanin had no effect by itself. When cells were preincubated (1 min) with galanin (10 f M–1 μ M), the CCK8-induced contraction was inhibited, with a maximal effect at 10 n M galanin. Likewise, VIP inhibited the CCK8-induced contraction with a maximal effect at 1 μ M. Preincubation of cells with somatostatin, N- ethylmaleimide , and (R)-p- cAMPS inhibited galanin- and VIP-induced relaxation. In conclusion, galanin induces a contraction of longitudinal smooth muscle cells that is dependent on an influx of extracellular calcium and an activation of a pertussis toxin G-protein. Galanin induces a relaxation of circular smooth muscle cells by activation of the adenylate cyclase complex and protein kinase A.

Effect of loxiglumide (CR 1505) on CCK-induced contractions and 3H-acetylcholine release from guinea-pig gallbladder

Release of [ 3H]-acetylcholine ( 3H-ACh) and muscle contractions in response to cholecystokinin (CCK) were measured and recorded simultaneously from isolated guinea-pig gallbladder. Cholecystokinin octapeptide (CCK8) (10 −10–10 −7 M) enhanced the release of [ 3H]ACh and the contractions of the muscle. TTX (10 −6 M) inhibited the CCK-induced release of 3H-ACh by only 30%. In Ca 2+-free medium CCK8 had no effect. Loxiglumide, (CR 1505), a newly synthesized nonpeptide CCK-A-receptor antagonist, D.L.- (3,4-dichlorbenzoilamino) -5 - / N - (3-methoxypropyl)-pentylamino-5-oxo-pentanoic acid, antagonized both the ACh-releasing effect of CCK and the contractions in a dose-dependent manner. The affinity (pA 2) of CR 1505 to CCK-receptors, determined by the shift of the concentration-response curves for CCK8 was 8.36. It was 5 logarithmic orders higher than the pA 2 of proglumide. The IC 50 value of CR 1505 calculated by the CCK-induced release of 3H-ACh was 10 nM. The results suggest the existence not only of muscular CCK receptors but also neuronal receptors for CCK probably located on cholinergic nerves.

Intracellular pathways triggered by galanin to induce contraction of pig ileum smooth muscle cells.

1. In order to determine the intracellular mechanisms by which galanin induces contraction of isolated smooth muscle cells from pig ileum, we examined the effects of external Ca2+, relaxing agents, pertussis toxin and forskolin on the galanin-induced contraction and compared these effects to those observed on the cholecystokinin derivative CCK8-induced contraction. 2. Galanin induced a concentration-dependent cell contraction. The maximal contraction (24.5 +/- 2.1% of the length of resting cells) was observed at 1 nM of galanin. When cells were incubated in the simultaneous presence of concentrations of galanin (10 fM) and CCK8 (1 pM) which were ineffective alone, or galanin (10 fM) and acetylcholine (100 pM), a synergistic action was observed corresponding to a submaximal contraction. 3. Incubation of cells in Ca(2+)-free medium caused a significant decrease in galanin- but not in CCK-induced contraction. Nifedipine, a Ca2+ channel blocker, provoked a concentration-dependent inhibition of galanin-induced contraction while it had no effect on the contraction induced by CCK8. 4. Vasoactive intestinal polypeptide (VIP) and isoprenaline, known to induce cell relaxation through an increase in intracellular cAMP level, inhibited CCK-induced cell contraction at concentrations ranging from 1 pM to 1 microM but failed to inhibit cell contraction induced by galanin. 5. When cells were pre-incubated for 3 h in the presence of 200 ng/ml of pertussis toxin, the contraction induced by galanin was abolished while the CCK-induced contraction remained unchanged. On the contrary, 10 microM forskolin abolished the contraction induced by 10 nM CCK but had no effect on galanin-induced contraction. 6. These results indicate that galanin induces a concentration-dependent contraction of pig ileum smooth muscle by a direct myogenic effect. This effect of galanin involves the activation of a pertussis toxin-sensitive G protein, which results in an influx of Ca2+ into the cell. This intracellular pathway is insensitive to the relaxing effect of cAMP.

Synthesis and biological activity of CCK peptides with antagonist activity for CCK-A-receptor

The synthesis of a series of cholecystokinin analogues derived from the well-known antagonist Ge 410 (Suc-Tyr(SO 3)-Met-Gly-Trp-Met-Asp-β-phenethylamide) is reported. Replacements of l-Trp by d-Trp, Asp by Glu and Met by Nle were carried out and the resulting changes in biological activities investigated. All compounds synthesized were tested for their ability to inhibit CCK-induced contraction on isolated guinea pig ileum. SAR studies for these compounds are discussed.

Enkephalin is a competitive antagonist of cholecystokinin in the gastrointestinal tract, as predicted from prior conformational analysis.

Prior calculations based on ECEPP (Empirical Conformational Energies for Peptides Program) of the low energy minima for cholecystokinin (CCK) and Met-enkephalin have demonstrated that significant structural features of these two peptides are identical. This result suggested the possibility that Met-enkephalin, as well as other enkephalin analogues of similar structure, could associate with receptors for CCK. To test this theoretical result, we examined the ability of Met-enkephalin and its analogues to bind to peripheral CCK receptors in the rat gastrointestinal tract; in particular, we measured the ability of the opiate peptide to inhibit the effects of CCK in a physiological assay system which we have previously characterized: CCK-induced contraction of the isolated rat pyloric sphincter. We find that Met-enkephalin is an antagonist of the CCK-8-induced contraction, with a IC50 of 110 nM. Furthermore, antibodies against CCK were found to cross-react with Met-enkephalin and its analogues in a manner which suggests a distinct structure-activity relationship. These experimental results strongly support the theoretical results of conformational analysis showing structural similarity between enkephalin and CCK. They further suggest that enkephalins could modulate the response of CCK systems under physiological conditions.

VIP inhibits CCK-induced gallbladder contraction involving a β-adrenoceptor mediated pathway in the rainbow trout, Oncorhynchus mykiss, in vivo

The effects of VIP on CCK-induced contractions of the gallbladder were studied in the rainbow trout, Oncorhynchus mykiss, in vivo. Gallbladder pressures were measured via a catheter inserted into the gallbladder, and drugs were injected through a catheter in the dorsal aorta. CCK-induced contractions of the gallbladder wall were reduced in a dose-dependent manner by VIP. The effect of VIP was inhibited by the β-adrenergic antagonist sotalol, indicating an adrenergic link in the VIP inhibition of the CCK-induced contraction. It is suggested that activation of VIP nerves could facilitate postprandial filling/refilling of the gallbladder despite the increased plasma levels of CCK during this period.

Involvement of calcium and IP3 in the CCK-induced contraction of monkey iris

Involvement of calcium and IP3 in the CCK-induced contraction of monkey iris

The influence of bile salts on small intestinal motility in the guinea pig in vitro

The effect of bile salts on intestinal motility is unclear. In the current study, isometric contractions of the guinea pig terminal ileum were examined in vitro. Dose-response curves to known agonists cholecystokinin (CCK), bethanechol, and KCl were constructed alone and in the presence of atropine (10−6 |mol/L), tetrodotoxin (10−6 mol/L), and different bile salts, namely, taurodeoxycholate, tauroursodeoxycholate, taurocholate, glycodeoxycholate, and glycoursodeoxycholate. These bile salts, at levels as low as 5 and 50 μmol/L, significantly depressed (P < 0.05) CCK-induced contractions throughout the dose-response curves and were concentration dependent. This depressant effect was not dependent on the bile salt species or any apparent physicochemical differences between them. The inhibitory effect was also specific for certain agonists such as CCK (the action of which was partially mediated by cholinergic nerves, being depressed by atropine and abolished by tetrodotoxin), field stimulation, and nicotine. Bile salts had no effect on either bethanechol- or KCl-induced contractions. Such bile salt inhibition of excitatory, cholinergic, enteric neurons may slow transit through the ileum, enhancing the time for absorption and conserving the bile salt pool.