Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 379, Issue 4, pp 389–395 | Cite as

Suitability of muscarinic acetylcholine receptor antibodies for immunohistochemistry evaluated on tissue sections of receptor gene-deficient mice

  • Gitte Jositsch
  • Tamara Papadakis
  • Rainer V. Haberberger
  • Miriam Wolff
  • Jürgen Wess
  • Wolfgang KummerEmail author
Short Communication


Acetylcholine (ACh) is a major regulator of visceral function exerting pharmacologically relevant effects upon smooth muscle tone and epithelial function via five types of muscarinic receptors (M1R-M5R). In this paper, we assessed the specificity of muscarinic receptor (MR) antibodies in immunohistochemical labelling on tissue sections by analysing specimens from wild-type and respective gene-deficient mice. Of 24 antibodies evaluated in this study, 16 were tested at 18 different conditions each, and eight of them in 21 different protocols, resulting in a total number of 456 antibody/protocol combinations. Each of them was tested at four antibody dilutions at minimum, so that finally, at least 1,824 conditions were evaluated. For each of them, dorsal root ganglia, urinary bladder and cross-sections through all thoracic viscera were investigated. In all cases where the antigen was available, at least one incubation condition was identified in which only select cell types were immunolabelled in the positive control but remained unlabelled in the pre-absorption control. With two exceptions (M2R antibodies), however, all antibodies produced identical immunohistochemical labelling patterns in tissues taken from corresponding gene-deficient mice even when the pre-absorption control in wild-type mice suggested specificity. Hence, the present data demonstrate the unpleasant fact that reliable immunohistochemical localisation of MR subtypes with antibodies is the exception rather than the rule. Immunohistochemical detection of MR subtype localisation in tissue sections of peripheral organs is limited to the M2R subtype utilising the most commonly used methodological approaches.


Acetylcholine Muscarinic receptor Immunohistochemistry Specificity Receptor knockout strains Mouse 



We thank Ms K. Michael for skilful help in preparing the figures. Thus study was supported by the DFG (Excellence Cluster Cardio-Pulmonary System and KO 1398/5-1).


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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Gitte Jositsch
    • 1
  • Tamara Papadakis
    • 1
  • Rainer V. Haberberger
    • 2
  • Miriam Wolff
    • 1
  • Jürgen Wess
    • 3
  • Wolfgang Kummer
    • 1
    Email author
  1. 1.Institute of Anatomy and Cell BiologyJustus-Liebig-University GiessenGiessenGermany
  2. 2.Department of Anatomy & HistologyFlinders University of South AustraliaAdelaideAustralia
  3. 3.Laboratory of Bioorganic ChemistryNational Institute of Diabetes and Digestive and Kidney DiseasesBethesdaUSA

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