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Neurochemical pattern of the complex innervation of neuroepithelial bodies in mouse lungs

  • Inge Brouns
  • Fusun Oztay
  • Isabel Pintelon
  • Ian De Proost
  • Robrecht Lembrechts
  • Jean-Pierre Timmermans
  • Dirk AdriaensenEmail author
Original Paper

Abstract

As best characterized for rats, it is clear that pulmonary neuroepithelial bodies (NEBs) are contacted by a plethora of nerve fiber populations, suggesting that they represent an extensive group of multifunctional intraepithelial airway receptors. Because of the importance of genetically modified mice for functional studies, and the current lack of data, the main aim of the present study was to achieve a detailed analysis of the origin and neurochemical properties of nerve terminals associated with NEBs in mouse lungs. Antibodies against known selective markers for sensory and motor nerve terminals in rat lungs were used on lungs from control and vagotomized mice of two different strains, i.e., Swiss and C57-Bl6. NEB cells were visualized by antibodies against either the general neuroendocrine marker protein gene-product 9.5 (PGP9.5) or calcitonin gene-related peptide (CGRP). Thorough immunohistochemical examination of NEB cells showed that some of these NEB cells also exhibit calbindin D-28 k (CB) and vesicular acetylcholine transporter (VAChT) immunoreactivity (IR). Mouse pulmonary NEBs were found to receive intraepithelial nerve terminals of at least two different populations of myelinated vagal afferents: (1) Immunoreactive (ir) for vesicular glutamate transporters (VGLUTs) and CB; (2) expressing P2X2 and P2X3 ATP receptors. CGRP IR was seen in varicose vagal nerve fibers and in delicate non-vagal fibers, both in close proximity to NEBs. VAChT immunostaining showed very weak IR in the NEB-related intraepithelial vagal sensory nerve terminals. nNOS- or VIP-ir nerve terminals could be observed at the base of pulmonary NEBs. While a single NEB can be contacted by multiple nerve fiber populations, it was clear that none of the so far characterized nerve fiber populations contacts all pulmonary NEBs. The present study revealed that mouse lungs harbor several populations of nerve terminals that may selectively contact NEBs. Although at present the physiological significance of the innervation pattern of NEBs remains enigmatic, it is likely that NEBs are receptor–effector end-organs that may host complex and/or multiple functional properties in normal airways. The neurochemical information on the innervation of NEBs in mouse lungs gathered in the present study will be essential for the interpretation of upcoming functional data and for the study of transgenic mice.

Keywords

Sensory airway receptors Myelinated vagal afferents C-fibers ATP receptors Glutamate NEBs Airways 

Notes

Acknowledgments

This work was supported by the following research grants: Fund for Scientific Research-Flanders (G.0085.04 and G.0081.08 to D.A.), NOI-BOF 2003 and GOA-BOF 2007 (to D.A.), and KP-BOF 2006 (to I·B.) from the University of Antwerp. We are grateful to Prof. G. Burnstock (Royal Free & University College Medical School) for his invaluable input in the ATP receptor studies, and acknowledge J. Van Genechten for his help with the study of the nitrergic innervation of mouse NEBs. We thank R. Spillemaeckers, G. Svensson, F. Terloo, S. De Geyter and G. Vermeiren for technical assistance, J. Van Daele and D. De Rijck for help with microscopy, imaging and illustrations, D. Vindevogel for aid with the manuscript, and H. De Pauw and S. Kockelberg for secretarial help.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Inge Brouns
    • 1
  • Fusun Oztay
    • 2
  • Isabel Pintelon
    • 1
  • Ian De Proost
    • 1
  • Robrecht Lembrechts
    • 1
  • Jean-Pierre Timmermans
    • 1
  • Dirk Adriaensen
    • 1
    Email author
  1. 1.Laboratory of Cell Biology and Histology, Department of Veterinary SciencesUniversity of AntwerpAntwerpBelgium
  2. 2.Department of Biology, Science FacultyIstanbul UniversityIstanbulTurkey

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