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Signal transduction pathway of the muscarinic receptors mediating gallbladder contraction

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Abstract

In gallbladder smooth muscle, carbachol interacts with M3 receptors to mediate contraction. To examine components of the intracellular second messenger system that is coupled to these receptors we have tested whether carbachol stimulates the formation of inositol phosphates (IP) to cause contraction. Guinea pig gallbladder muscle strips were prelabeled with [3H]inositol and were incubated with 0.1 mmol/l carbachol, a concentration causing maximal contraction. [3H]inositol monophosphates, [3H]inositol bisphosphates and [3H]inositol trisphosphates and contraction were measured at various times (0–90 s). To examine whether a pertussis toxin-sensitive guanine nucleotide binding protein is coupled to the muscarinic receptors, guinea pigs were pretreated with pertussis toxin (180 μg/kg i.v./24 h). The effectiveness of pertussis toxin treatment was determined by measuring [32P]ADP-ribosylation of a −40/41 kDa protein from gallbladder homogenates. Carbachol caused a significant time-dependent increase in the formation of [3H]inositol monophosphates, [3H]inositol bisphosphates and [3H]inositol trisphosphates. The time course of [3H]inositol trisphosphate turnover caused by carbachol was biphasic, and was detectable at 15 s and maximal at 60 s; at 75 s and 90 s formation of [3H]inositol trisphosphates decreased, whereas the time course of carbachol-induced contraction of the gallbladder smooth muscle strips reached a plateau after 90 s. The effects of carbachol on [3H]inositol trisphosphates and on contraction were abolished by atropine. Pretreatment with pertussis toxin resulted in ADP-ribosylation of a ≈40/41 kDa protein from gallbladder cell membranes but did not affect the concentration-response or time course of carbachol-induced contraction. These results indicate that carbachol-induced contraction of gallbladder smooth muscle cells is accompanied by the activation of inositol phosphate turnover and does not involve a pertussis toxin-sensitive G-protein.

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Authors and Affiliations

  1. Medizinische Kernklinik und Poliklinik, Universitäts-Krankenhaus Eppendorf, Martinistrasse 52, D-20246, Hamburg, Germany

    Tammo von Schrenck, Burkhard Mackensen, Jörg Sievers, Silvia Mirau, Andreas Raedler & Heiner Greten

  2. Abteilung Allgemeine Pharmakologie, Universitäts-Krankenhaus Eppendorf, Martinistrasse 52, D-20246, Hamburg, Germany

    Ulrike Mende & Wilhelm Schmit

Authors
  1. Tammo von Schrenck
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  2. Burkhard Mackensen
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  3. Ulrike Mende
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  4. Wilhelm Schmit
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  5. Jörg Sievers
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  6. Silvia Mirau
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  7. Andreas Raedler
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  8. Heiner Greten
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Additional information

This article is based in part on the doctoral thesis of Burkhard Mackensen at the Faculty of Medicine, University of Hamburg, Germany. Some of the results were presented at the meeting of the American Gastroenterological Association (AGA) in San Francisco 1992 (von Schrenck et al. 1992)

Correspondence to: T. von Schrenck at the above address

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von Schrenck, T., Mackensen, B., Mende, U. et al. Signal transduction pathway of the muscarinic receptors mediating gallbladder contraction. Naunyn-Schmiedeberg’s Arch Pharmacol 349, 346–354 (1994). https://doi.org/10.1007/BF00170879

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  • DOI: https://doi.org/10.1007/BF00170879

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