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The detection of the non-M2 muscarinic receptor subtype in the rat heart atria and ventricles

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Abstract

Mammal heart tissue has long been assumed to be the exclusive domain of the M2 subtype of muscarinic receptor, but data supporting the presence of other subtypes also exist. We have tested the hypothesis that muscarinic receptors other than the M2 subtype are present in the heart as minor populations. We used several approaches: a set of competition binding experiments with pirenzepine, AFDX-116, 4-DAMP, PD 102807, p-F-HHSiD, AQ-RA 741, DAU 5884, methoctramine and tripinamide, blockage of M1 muscarinic receptors using MT7 toxin, subtype-specific immunoprecipitation experiments and determination of phospholipase C activity. We also attempted to block M1–M4 receptors using co-treatment with MT7 and AQ-RA 741. Our results show that only the M2 subtype is present in the atria. In the ventricles, however, we were able to determine that 20% (on average) of the muscarinic receptors were subtypes other than M2, with the majority of these belonging to the M1 subtype. We were also able to detect a marginal fraction (6 ± 2%) of receptors that, based on other findings, belong mainly to the M5 muscarinic receptors. Co-treatment with MT7 and AQ-RA 741 was not a suitable tool for blocking of M1–M4 receptors and can not therefore be used as a method for M5 muscarinic receptor detection in substitution to crude venom. These results provide further evidence of the expression of the M1 muscarinic receptor subtype in the rat heart and also show that the heart contains at least one other, albeit minor, muscarinic receptor population, which most likely belongs to the M5 muscarinic receptors but not to that of the M3 receptors.

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Acknowledgements

We would like to thank Prof. Lincoln Potter from University of Miami for his advice concerning MT7 and M5 muscarinic receptor detection, Dr. Maria-Laura Bolognesi from the University of Bologna for her advice with tripinamide properties, Prof. C. Melchiorre from the same University for his generous gift of this compound and Prof. K. Ladinsky from the Istituto De Angeli, S.p.A for his generous gift of compound AFDX-116. We appreciate the gift of AQ-RA 741 from Boehringer Ingelheim, Biberach, Germany (Dr. Doods and Dr. Mueller). The gift of PD 102807 from Parke-Davis is greatly appreciated. The work was supported mainly by grant GAUK 519/07 from Grant Agency of Charles University and in part by grant GAUK 289/07 from the same Grant Agency. Another support was obtained from grants MSMT CR LC554, MSM0021620849, GACR 304/08/0256 and by the Research project AV0Z 50110509. We gratefully acknowledge the technical assistance by Mrs. Dana Ungerová, Romana Ondřejová, Olga Martínková and Eva Šimáková.

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  1. Institute of Physiology, 1st Faculty of Medicine, Charles University, Albertov 5, 12800, Prague, Czech Republic

    Jaromir Myslivecek, Martin Klein & Martina Novakova

  2. Prague Psychiatric Centre, Ustavni 91, 181 03, Prague, Czech Republic

    Jan Ricny

  3. Institute of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, 14220, Prague, Czech Republic

    Jan Ricny

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  1. Jaromir Myslivecek
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  2. Martin Klein
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Correspondence to Jaromir Myslivecek.

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Myslivecek, J., Klein, M., Novakova, M. et al. The detection of the non-M2 muscarinic receptor subtype in the rat heart atria and ventricles. Naunyn-Schmied Arch Pharmacol 378, 103–116 (2008). https://doi.org/10.1007/s00210-008-0285-8

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