Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver

Elsing, Christoph; Hübner, Christian A.; Fitscher, B.A.; Kassner, Astrid; Stremmel, Wolfgang

doi:10.1002/hep.510250404

Cited by 25 publications

(20 citation statements)

References 48 publications

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“…The 5Ј sequence of the primers started at base pair 997 (5Ј-GGAGTCCCTCACATCCTC-CGA-3Ј) and the 3Ј sequence started at base pair 1471 (5Ј-CGC-CAGCGCCTCTTGTCCCAG-3Ј) with an expected fragment length of 474 bp. For the molecular analysis of the M1 muscarinic ACh receptor in pure preparations of cholangiocytes, we used the same RT-PCR conditions as Elsing et al (37): 30 cycles of denaturation at 95ЊC for 1 min annealing at 65ЊC for 1 min, and extension at 72ЊC for 1 min. Rat brain and yeast transfer RNA were used as positive and negative controls, respectively.…”

Section: In Vitro Molecular Analysismentioning

confidence: 99%

Role and mechanisms of action of acetylcholine in the regulation of rat cholangiocyte secretory functions.

Alvaro¹,

Alpini²,

Jezequel³

et al. 1997

J. Clin. Invest.

125

159

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We investigated, in isolated bile duct units (IBDU) and cholangiocytes isolated from normal rat liver, the occurrence of acetylcholine (ACh) receptors, and the role and mechanisms of ACh in the regulation of the Cl Ϫ /HCO 3 Ϫ exchanger activity. The Cl Ϫ /HCO 3 Ϫ exchanger activity was evaluated measuring changes in intracellular pH induced by acute Cl Ϫ removal/readmission. M3 subtype ACh receptors were detected in IBDU and isolated cholangiocytes by immunofluorescence, immunoelectron microscopy, and reverse transcriptase PCR. M1 subtype ACh receptor mRNA was not detected by reverse transcriptase PCR and M2 subtype was negative by immunofluorescence. ACh (10 M) showed no effect on the basal activity of the Cl Ϫ /HCO 3 Ϫ exchanger. When IBDU were exposed to ACh plus secretin, ACh significantly ( P Ͻ 0.03) increased the maximal rate of alkalinization after Cl Ϫ removal and the maximal rate of recovery after Cl

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Section: In Vitro Molecular Analysismentioning

confidence: 99%

Role and mechanisms of action of acetylcholine in the regulation of rat cholangiocyte secretory functions.

Alvaro¹,

Alpini²,

Jezequel³

et al. 1997

J. Clin. Invest.

125

159

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“…5 IP 3 -dependent Ca 2ϩ signals stimulate bile canalicular contractions, an effect blocked by nitric oxide. 6 In intrahepatic bile duct epithelial cells, stimulation of purinergic receptors with adenosine triphosphate results in Ca 2ϩ signals and increases membrane Cl Ϫ and K ϩ permeabilities 7 ; stimulation of the muscarinic acetycholine receptor triggers IP 3 -dependent Ca 2ϩ oscillations 8 and enhances the effect of secretin on the Cl Ϫ /HCO 3 Ϫ exchanger. 9 Three isoforms of IP 3 R have been identified in humans and rodents.…”

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confidence: 99%

Expression of inositol 1,4,5-trisphosphate receptor isoforms in rat cirrhosis

et al. 1999

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Ca 2؉ signals mediate the hepatic effects of numerous hormones and growth factors. Hepatic Ca 2؉ signals are elicited by the inositol trisphosphate receptor, an intracellular Ca 2؉ channel. Three isoforms of this receptor have been identified; they are expressed and regulated differently. We investigated the effect of liver fibrosis and cirrhosis on the hepatic expression of the inositol trisphosphate receptor isoforms. Two different rat models were used: bile duct ligation (fibrosis) and chronic exposure to CCl 4 /phenobarbital (cirrhosis). Messenger RNA levels were determined by ribonuclease protection assay (RPA), competitive polymerase chain reaction (PCR) followed by Southern blotting, and real-time quantitative PCR. Protein expression was assessed by Western blotting; tissue distribution was assessed by immunohistology. In control animals, isoform 2 was the predominant isoform, isoform 1 represented less than one third, and isoform 3 less than 1%. After bile duct ligation, expression of types 1 and 3 increased 1.9-and 5.7-fold, and expression of type 2 decreased 2.5-fold at the protein level. After exposure to CCl 4 /phenobarbital, expression of types 1, 2, and 3 were 2.4-, 0.9-, and 4.2-fold their expression in control animals. Type 2 was localized to the apical domain of hepatocytes, consistent with a role for Ca 2؉ signals in canalicular function. Type 3 was detectable in intrahepatic bile duct epithelial cells and not in hepatocytes, suggesting that Ca 2؉ signals may be regulated differently in these cells. Signaling through inositol trisphosphate receptor participates in the pathogenesis of cirrhosis, because this process affects the expression of its isoforms. (HEPATOLOGY 1999;30:1018-1026.)Development of cirrhosis implies not only remodeling of the liver architecture, but also dramatic changes in the functions of hepatic cells. These changes are orchestrated by a network of receptors, protein-kinases, and transducers that compose complex signaling pathways. Among them, the inositol 1,4,5-trisphosphate receptor (IP 3 R) plays a central role. 1 Numerous hormones and growth factors bind to receptors on plasma membrane to stimulate the production of inositol 1,4,5-trisphosphate (IP 3 ) and to trigger IP 3 -dependent Ca 2ϩ release through the Ca 2ϩ channel of the IP 3 R. 2 Ca 2ϩ signals regulate many cellular functions from gene expression and cellular division to metabolic pathways and cellular secretion. 1 Both hepatocytes and biliary epithelial cells demonstrate complex Ca 2ϩ signals, which rely essentially on IP 3 R, because the ryanodine receptor, another intracellular Ca 2ϩ channel, could not be detected in liver. 3 In response to hormonal stimulation, intracellular Ca 2ϩ concentration oscillates in hepatocytes and even forms Ca 2ϩ waves through the entire lobule. 4 Hepatocellular Ca 2ϩ signals start in a specific apical location and diffuse in an apical to basal manner. 5 IP 3 -dependent Ca 2ϩ signals stimulate bile canalicular contractions, an effect blocked by nitric oxide. 6 In intrahepatic bil...

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“…Isolated guinea pig liver perfusions were performed as described. 17 Isotonic Krebs-Henseleit bicarbonate buffer contained (in mmol/L) NaCl 120, KCl 5, CaCl 2 1.25, MgSO 4 0.65, NaHCO 3 25, KH 2 PO 4 1.17, glucose 5. Krebs-Henseleit bicarbonate buffer was gassed continuously with a mixture of 95% O 2 and 5% CO 2 and maintained at 37.0 Ϯ 1.0°C and pH 7.40 Ϯ 0.5.…”

Section: Methodsmentioning

confidence: 99%

“…To investigate acid-base carriers and intracellular cyclic adenosine monophosphate (cAMP), guinea pig BEC were isolated as previously described for isolation of rat BEC. 15,17 Livers were first perfused for 15 minutes with a gazed buffer of 0.2 mmol/L ethyleneglycol-bis-(␤-aminoethyl ether)-N-N tetraacetic acid, 0.143 mol/L NaCl, 7 mmol/L KCl, and 10 mmol/L HEPES-NaOH (pH 7.4). This medium was then replaced by a medium containing 0.1 mol/L NaCl, 7 mmol/L KCl, 5 mmol/L CaCl 2 , 50 mmol/L HEPES-NaOH (pH 7.6), 0.1% collagenase, and 0.1% hyaluronidase.…”

Section: Methodsmentioning

confidence: 99%

Sodium, Hydrogen exchange type 1 and bile ductular secretory activity in the guinea pig

2000

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Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver

Cited by 25 publications

References 48 publications

Role and mechanisms of action of acetylcholine in the regulation of rat cholangiocyte secretory functions.

Role and mechanisms of action of acetylcholine in the regulation of rat cholangiocyte secretory functions.

Expression of inositol 1,4,5-trisphosphate receptor isoforms in rat cirrhosis

Sodium, Hydrogen exchange type 1 and bile ductular secretory activity in the guinea pig

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