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Neuropeptides in intact and denervated tympanic membranes

An immunohistofluorescence study in the rat

Archives of oto-rhino-laryngology volume 246, pages 183–189 (1989)Cite this article

Summary

The distribution and origin of peptidecontaining and sympathetic nerve fibers were studied in the tympanic membrane of rats with intact innervation, and in rats following sympathectomy, vagotomy, or capsaicin treatment. Nerve fibers showing substance-P-like immunoreactivity (SP-LI), calcitonin gene-related peptide (CGRP)-LI, vasoactive intestinal polypeptide (VIP)-LI, enkephalin-LI, neuropeptide Y (NPY)-LI and tyrosine hydroxylase (TH)-LI were detected along blood vessels in the pars flaccida, the external auditory canal and in the fibrocartilaginous ring of the pars tensa. In the pars flaccida there were numerous fibers demonstrating SP-LI and CGRP-LI, while there were few such fibers in the pars tensa. In both portions of the tympanic membrane these fibers were present within and beneath the keratinized stratified squamous epithelium. In the pars flaccida, nerve fibers showing SP-LI and CGRP-LI were also seen near mast cells. Sympathectomy led to a loss of nerve fibers showing TH-LI and NPY-LI, whereas the other peptide-containing nerve fibers remained unaffected. Vagotomy did not reduce the immunoreactivity for any of the neuropeptides studied. Capsaicin treatment caused a reduction in nerve fibers displaying SP-LI and CGRP-LI. The abundance of nerve fibers showing SP-LI and CGRP-LI in the keratinized squamous epithelium indicates that the tympanic membrane is richly supplied with sensory nerves. The localization of nerve fibers exhibiting these latter substances in the vicinity of mast cells in the pars flaccida suggests that this part of the tympanic membrane is a site where neurogenic inflammation occurs.

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References

  1. Albiin N, Hellström S, Salén B, Stenfors L-E, Wirell S (1985) The vascular supply of the rat tympanic membrane. Anat Rec 212:17–22

    Google Scholar 

  2. Alm PE, Bloom GD, Hellström S, Stenfors L-E, Widemar L (1983) Mast cells in the pars flaccida of the tympanic membrane. A quantitative, morphological and biochemical study in the rat. Experientia 39:287–289

    Google Scholar 

  3. Bienenstock J, Tomioka M, Matsuda H, Stead RH, Quinonez G, Simon GT, Coughlin MD, Denburg JA (1987) The role of mast cells in inflammatory processes: evidence for nerve/mast cell interactions. Int Arch Allergy Appl Immunol 82:238–243

    Google Scholar 

  4. Brain SD, Williams TJ, Tippins JR, Morris HR, MacIntyre I (1985) Calcitonin gene-related peptide is a potent vasodilator. Nature 313:54–56

    Google Scholar 

  5. Dubowitz V, Brooke MH (1973) Muscle biopsy: a modern approach. Saunders, London, pp 20–23

    Google Scholar 

  6. Foreman JC (1987) Peptides and neurogenic inflammation. Br Med Bull 43:386–400

    Google Scholar 

  7. Foreman JC, Jordan CC, Oehme P, Renner H (1983) Structure-activity relationships for some substance P-related peptides that cause wheal and flare reaction in the human skin. J Physiol 335:449–465

    Google Scholar 

  8. Forsgren S, Söderberg L (1987) Immunohistochemical procedures for the demonstration of peptide- and tyrosine hydroxylase containing nerve fibers in cryostat sections of unfixed rapidly frozen tissue stored for long periods of time. Histochemistry 87:561–568

    Google Scholar 

  9. Fritz P, Müller J, Reiser, H, Saal SG, Hadam M, Tuezek HV, Wegner G, Laschner W (1986) Avidin-peroxidase. A new mast cell staining method. Acta Histochem (Jena) 32:235–239

    Google Scholar 

  10. Goldie P, Hellström S (1988) Autonomic nerves and middle ear fluid production. An experimental study in the rat. Acta Otolaryngol (Stockh) 106:10–18

    Google Scholar 

  11. Goldie P, Hellstöm S, Idahl L-Å (1989) Middle ear effusion induced by various inflammatory mediators and neuropeptides. An experimental study in the rat. Acta Otolaryngol (Stockh) (in press)

  12. Hellström S, Salén B, Stenfors L-E (1982) The site of initial production and transport of effusion materials in otitis media serosa. Acta Otolaryngol (Stockh) 93:435–440

    Google Scholar 

  13. Lundberg JM, Hökfelt T, Kewenter J, Pettersson G, Ahlman H, Edin R, Dahlström A, Nilsson G, Terenius L, Uvnäs-Wallensten K, Said S (1979) Substance P-, VIP-, and enkephalin-like immunoreactivity in the human vagus nerve. Gastroenterology 77:468–471

    Google Scholar 

  14. Lundberg JM, Terenius L, Hökfelt T, Martling C-M, Tatemoto K, Mutt V, Polak J, Bloom SR (1982) Neuropeptide Y (NPY)-like immunoreactivity in peripheral noradrenergic neurons and effects of NPY on sympathetic function. Acta Physiol Scand 116:477–480

    Google Scholar 

  15. Lundberg JM, Torsell L, Sollevi A, Pernow J, Theodorsson Norheim E, Änggård A, Hamberger B (1985) Neuropeptide Y and sympathetic vascular control in man. Regul Pept 13:41–52

    Google Scholar 

  16. Lundberg JM, Lundblad L, Martling C-R, Saria A, Stjärne P, Änggård A (1987) Coexistence of multiple peptides and classic transmitters in airway neurons: functional and pathophysiologic aspects. Am Rev Respir Dis 136:S16-S22

    Google Scholar 

  17. Mulderry PK, Ghatei MA, Rodrigo J, Allen JM, Rosenfeld MG, Polak JM, Bloom SR (1985) Calcitonin gene-related peptide in cardiovascular tissues of the rat. Neuroscience 14:947–954

    Google Scholar 

  18. Murphy R, Furness JB, Beardsley AM, Costa M (1982) Characterisation of substance P-like immunoreactivity in peripheral sensory nerves and enteric nerves by high pressure liquid chromatography and radioimmunoassay. Regul Rept 4:203–212

    Google Scholar 

  19. Paparella MM (1980) Definition and classification. In: Proceedings of the Second International Symposium on Recent Advances in Otitis Media with Effusion. Ann Otorhinolaryngol (Suppl 68) 89:3–4

    Google Scholar 

  20. Pitrowski W, Foreman JC (1986) Some effects of calcitonin gene-related peptide in human skin and on histamine release. Br J Dermatol 114:37–46

    Google Scholar 

  21. Torre JS de la, Surgeon JW (1976) A methodological approach to rapid and sensitive monoamine histofluorescence using a modified glyoxylic acid technique: the SPG method. Histochemistry 49:81–93

    Google Scholar 

  22. Uddman R, Sundler F, Emson P (1984) Occurrence and distribution of neuropeptide-Y-immunoreactive nerves in the respiratory tract and middle ear. Cell Tissue Res 237:321–327

    Google Scholar 

  23. Uddman R, Luts A, Sundler F (1985) Occurrence and distribution of calcitonin gene-related peptide in the mammalian respiratory tract and middle ear. Cell Tissue Res 241:551–555

    Google Scholar 

  24. Uddman R, Edvinsson L, Ekblad E, Håkansson R, Sundler F (1986) Calcitonin gene-related peptide (CGRP): perivascular distribution and vasodilatory effects. Regul Pept 15:1–23

    Google Scholar 

  25. Uddman R, Grunditz T, Larsson A, Sundler F (1988) Sensory innervation of the ear drum and middle ear mucosa: retrograde tracing and immunocytochemistry. Cell Tissue Res 252:141–146

    Google Scholar 

  26. Wharton J, Gulbenkian S, Mulderry PK, Ghatei MA, McGregor GP, Bloom SR, Polak JM (1986) Capsaicin induces a depletion of calcitonin gene-related peptide (CGRP)-immunoreactive nerves in the cardiovascular system of the guinea pig and rat. J Auton Nerv Syst 16:289–309

    Google Scholar 

  27. Widemar L, Hellström S, Schultzberg U, Stenfors L-E (1985) Autonomic innervation of the tympanic membrane. An immunocytochemical and histofluorescence study. Acta Otolaryngol (Stockh) 100:58–65

    Google Scholar 

  28. William TJ (1982) Vasoactive intestinal polypeptide is more potent than prostaglandin E as a vasodilator and oedema potentiator in rabbit skin. Br J Pharmacol 77:505–509

    Google Scholar 

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Affiliations

  1. Department of Anatomy, University of Umeå, S-90187, Umeå, Sweden

    P. Goldie, S. Hellström & S. Forsgren

  2. Departments of Otorhinolaryngology & Head and Neck Surgery, University of Umeå, S-90187, Umeå, Sweden

    S. Hellström

Authors
  1. P. Goldie
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  2. S. Hellström
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  3. S. Forsgren
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Goldie, P., Hellström, S. & Forsgren, S. Neuropeptides in intact and denervated tympanic membranes. Arch Otorhinolaryngol 246, 183–189 (1989). https://doi.org/10.1007/BF00453658

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  • DOI: https://doi.org/10.1007/BF00453658

Key words

  • Tympanic membrane
  • Mast cells
  • Substance P
  • Calcitonin gene-related peptide
  • Autonomic nervous system