Skip to main content
Log in

Ultrastructural evidence for the coexistence of calcitonin gene-related peptide and substance P in secretory vesicles of peripheral nerves in the guinea pig

  • Published:
Journal of Neurocytology

Summary

Using double immunogold staining procedures, calcitonin gene-related peptide (CGRP)-like and substance P (SP)-like immunoreactivities were localized at the ultrastructural level to guinea pig trigeminal ganglia, dorsal root ganglia and peripheral nerve fibres associated with the vascular system. CGRP-like and SP-like immunoreactivities were found consistently in large granular secretory vesicles (70–100 nm in diameter), and both peptide immunoreactivities were co-localized to the same vesicle in both sensory ganglion cells and within axons and their terminals in the adventitia and adventitial-medial border of the superior mesenteric artery. These results suggest that CGRP and SP are co-stored and may be released together from peripheral axons in the guinea pig.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Amara, S. G., Arriza, J. L., Leff, S. E., Swanson, L. W., Evans, R. M. &Rosenfeld, M. G. (1985) Expression in brain of a messenger RNA encoding a novel neuropeptide homologous to calcitonin gene-related peptide.Science 229, 1094–7.

    Google Scholar 

  • Amara, S. G., Jonas, V., Rosenfeld, M. G., Ong, E. S. &Evans, R. M. (1982) Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products.Nature 298, 240–4.

    Google Scholar 

  • Baumgarten, H. G., Holstein, A. F. &Owman, C. H. (1970) Auerbach's plexus of mammals and man: electron microscopical identification of three different types of neuronal processes in myenteric ganglia of the large intestine from rhesus monkeys, guinea pigs and man.Zeitschrift für Zellforschung und mikroskopische Anatomie 106, 376–97.

    Google Scholar 

  • Bendayan, M. (1982) Double immunocytochemical labelling applying the protein A-gold technique.Journal of Histochemistry and Cytochemistry 30, 81–5.

    Google Scholar 

  • Brain, S. D. &Williams, T. J. (1985) Inflammatory oedema induced by synergism between calcitonin gene-related (CGRP) and mediators of increased vascular permeability.British journal of Pharmacology 86, 855–60.

    Google Scholar 

  • Brain, S. D., Williams, T. J., Tippins, J. R., Morris, H. R. &MacIntyre, I. (1985) Calcitonin gene-related peptide is a potent vasodilator.Nature 313, 54–6.

    Google Scholar 

  • Chan-Palay, V. &Palay, S. L. (1977) Ultrastructural identification of substance P cells and their processes in rat sensory ganglia and their terminals in the spinal cord by immunocytochemistry.Proceedings of the National Academy of Sciences USA 74, 4050–4.

    Google Scholar 

  • Cook, R. D. &Burnstock, G. (1976) The ultrastructure of Auerbach's plexus in the guinea pig. I. Neuronal elements.Journal of Neurocytology 5, 171–94.

    Google Scholar 

  • Edvinsson, L., McCulloch, J., Rosell, S. &Udman, R. (1982) Antagonism by (D-Pro2, D-Trp7,9)-substance P of the cerebrovascular dilatation induced by substance P.Acta physiologica scandinavica 116, 411–16.

    Google Scholar 

  • Fehér, E., Burnstock, G., Varndell, I. M. &Polak, J. M. (1986) Calcitonin gene-related peptide-containing nerve fibres in the small intestine of the guinea pig; electron microscopic immunocytochemistry.Cell and Tissue Research (in press).

  • Fischer, J., Forssman, W. G., Hökfelt, T., Lundberg, J. M., Reinecke, M., Tschopp, F. A. &Wiesenfeld-Hallin, Z. (1985) Immunoreactive calcitonin gene-related peptide and substance P: coexistence in sensory neurones and behavioural interaction after intrathecal administration in the rat.Journal of Physiology 362, 29P.

    Google Scholar 

  • Gamse, R. &Saria, A. (1985) Potentiation of tachykinin-induced plasma protein extravasation by calcitonin gene-related peptide.European Journal of Pharmacology 114, 61–6.

    Google Scholar 

  • Geuze, H. J., Slot, J. W., Scheffer, R. C. T. &Van Der Ley, P. A. (1981) Use of colloidal gold particles in double-labelling immunoelectron microscopy of ultrathin frozen tissue sections.Journal of Cell Biology 89, 653–65.

    Google Scholar 

  • Gibbins, I. L., Furness, J. B., Costa, M., MacIntyre, I., Hillyard, C. J. &Girgis, S. (1985) Co-localisation of calcitonin gene-related peptide-like immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs.Neuroscience Letters 57, 125–30.

    Google Scholar 

  • Gibson, S. J., Polak, J. M., Bloom, S. R., Sabate, I. M., Mulderry, P. M., Ghatei, M. A., McGregor, G. P., Morrison, J. F. B., Kelly, J. S., Evans, R. M. &Rosenfeld, M. G. (1984) Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and of eight other species.Journal of Neuroscience 4, 3101–11.

    Google Scholar 

  • Harmer, A., Schofield, J. G. &Keen, P. (1980) Cycloheximide-sensitive synthesis of substance P by isolated dorsal root ganglia.Nature 284, 267–9.

    Google Scholar 

  • Lee, Y., Kawai, Y., Shiosaka, S., Takami, K., Kiyama, H., Hillyard, C. J., Girgis, S., MacIntyre, I., Emson, P. C. &Tohyama, M. (1985) Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglia of the rat: immunohistochemical analysis.Brain Research 330, 194–6.

    Google Scholar 

  • Le Greves, P., Nyberg, F., Terenius, L. &Hökfelt, T. (1985) Calcitonin gene-related peptide is a potent inhibitor of substance P degradation.European Journal of Pharmacology 115, 309–11.

    Google Scholar 

  • Lundberg, J. M., Franco-Cereceda, A., Hua, X., Hökfelt, T. &Fischer, J. A. (1985) Coexistence of substance P- and calcitonin gene-related peptide-like immunoreactitivites in sensory nerves in relation to cardiovascular and bronchoconstrictor effects of capsaicin.European Journal of Pharmacology 108, 315–19.

    Google Scholar 

  • Mason, R. T., Peterfreund, R. A., Sawchenko, P. E., Corrigan, A. Z., Rivier, J. E. &Vale, W. W. (1984) Release of the predicted calcitonin gene-related peptide from cultured rat trigeminal ganglion cells.Nature 308, 653–5.

    Google Scholar 

  • Mulderry, P. K., Ghatei, M. A., Bishop, A. E., Allen, Y. S., Polak, J. M. &Bloom, S. R. (1985a) Distribution and chromatographic characterisation of CGRP-like immunoreactivity in the brain and gut of the rat.Regulatory Peptides 12, 133–43.

    Google Scholar 

  • Mulderry, P. K., Ghatei, M. A., Rodrigo, J., Allen, J. M., Rosenfeld, M. G., Polak, J. M. &Bloom, S. R. (1985b) Calcitonin gene-related peptide in cardiovascular tissues of the rat.Neuroscience 14, 947–54.

    Google Scholar 

  • Probert, L., De Mey, J. &Polak, J. M. (1981) Distinct subpopulation of enteric p-type neurones contain substance P and vasoactive intestinal polypeptide.Nature 294, 470–1.

    Google Scholar 

  • Probert, L., De Mey, J. &Polak, J. M. (1983) Ultrastructural localisation of four different neuropeptides within separate populations of p-type nerves in the guinea pig colon.Gastroenterology 85, 1094–104.

    Google Scholar 

  • Romano, E. L. &Romano, M. (1977) Staphylococcal protein A bound to colloidal gold: a useful reagent to label antigen-antibody sites in electron microscopy.Immunochemistry 14, 711–15.

    Google Scholar 

  • Rosenfeld, M. G., Mermod, J. J., Amara, S. G., Swanson, L. W., Rivier, J., Vale, W. W. &Evans, R. M. (1983) Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing.Nature 304, 129–35.

    Google Scholar 

  • Sikri, K. L., Hoyes, A. D., Barber, P. &Jagessar, H. (1981) Substance P-like immunoreactivity in the intramural nerve plexuses of the guinea pig ureter: a light and electron microscopical study.Journal of Anatomy 133, 425–42.

    Google Scholar 

  • Slot, W. J. &Geuze, H. T. (1984) Gold markers for single and double immunolabelling of ultrathin cryosections. InImmunolabelling for Electron Microscopy (edited byPolak, J. M. &Varndell, I. M.), pp. 129–42. Amsterdam: Elsevier Science Publishers.

    Google Scholar 

  • Tapia, F. J., Varndell, I. M., Probert, L., De Mey, J. &Polak, J. M. (1983) Double immunogold staining method for the simultaneous ultrastructural localisation of regulatory peptides.Journal of Histochemistry and Cytochemistry 31, 977–81.

    Google Scholar 

  • Terenghi, G., Polak, J. M., Ghatei, M. A., Mulderry, P. K., Butler, J. M., Unger, W. G. &Bloom, S. R. (1985) Distribution and origin of calcitonin gene-related peptide (CGRP) immunoreactivity in the sensory innervation of the mammalian eye.Journal of Comparative Neurology 233, 506–16.

    Google Scholar 

  • Terenghi, G., Polak, J. M., Probert, L., McGregor, G. P., Ferri, G. L., Blank, M. A., Butler, J. M., Unger, W. G., Zhang, S., Cole, D. F. &Bloom, S. R. (1982) Mapping, quantitative Distribution and origin of substance P- and VIP-Containing nerves in the uvea of guinea pig eye.Histochemistry 75, 399–417.

    Google Scholar 

  • Varndell, I. M., Tapia, F. J., Probert, L., Buchan, A. M. J., Gu, J., De Mey, J., Bloom, S. R. &Polak, J. M. (1982) Immunogold staining procedure for the localisation of regulatory peptides.Peptides 3, 259–72.

    Google Scholar 

  • Wang, B. L. &Larsson, L. I. (1985) Simultaneous demonstration of multiple antigens by indirect immunofluorescence or immunogold staining. Novel light and electron microscopical double and triple staining method employing primary antibodies from the same species.Histochemistry 83, 47–56.

    Google Scholar 

  • Wharton, J., Gulbenkian, S., Bloom, S. R. &Polak, J. M. (1985) The capsaicin-sensitive (afferent) innervation of the guinea pig cardiovascular system contains both calcitonin gene-related peptide (CGRP) and substance P (SP).Neuroscience Letters Suppl.22, S86.

    Google Scholar 

  • Wharton, J., Polak, J. M., McGregor, G. P., Bishop, A. E. &Bloom, S. R. (1981a) The distribution of substance P-like immunoreactive nerves in the guinea pig heart.Neuroscience 6, 2193–204.

    Google Scholar 

  • Wharton, J., Polak, J. M., Probert, L., De Mey, J., McGregor, G. P., Bryant, M. G. &Bloom, S. R. (1981b) Peptide-containing nerves in the ureter of the guinea pig and cat.Neuroscience 6, 969–82.

    Google Scholar 

  • Wiesenfeld-Hallin, Z., Hökfelt, T., Lundberg, J. M., Forssman, W. G., Reinecke, M., Tschopp, F. A. &Fischer, J. A. (1984) Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neurons in spinal cord and interact in spinal behavioural responses of the rat.Neuroscience Letters 52, 199–204.

    Google Scholar 

  • Yaksh, T. L., Jessell, T. M., Gamse, R., Mudge, A. W. &Leeman, S. E. (1980) Intrathecal morphine inhibits substance P release from mammalian spinal cordin vivo.Nature 286, 155–57.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gulbenkian, S., Merighi, A., Wharton, J. et al. Ultrastructural evidence for the coexistence of calcitonin gene-related peptide and substance P in secretory vesicles of peripheral nerves in the guinea pig. J Neurocytol 15, 535–542 (1986). https://doi.org/10.1007/BF01611735

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01611735

Keywords

Navigation