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Extrinsic and intrinsic sources of calcitonin gene-related peptide immunoreactivity in the lamb ileum: a morphometric and neurochemical investigation

Cell and Tissue Research volume 323, pages 183–196 (2006)Cite this article

Abstract

To investigate extrinsic origins of calcitonin gene-related peptide immunoreactive (CGRP-IR) nerve fibres in the sheep ileum, the retrograde fluorescent tracer Fast Blue (FB) was injected into the ileum wall. Sections of thoraco-lumbar dorsal root ganglia (DRG) and distal (nodose) vagal ganglia showing FB-labelled neurons were processed for CGRP immunohistochemistry. The distribution of CGRP-IR in fibres and nerve cell bodies in the ileum was also studied. CGRP-IR enteric neurons were morphometrically analysed in myenteric (MP) and submucosal plexuses (SMP) of lambs (2–4 months). Sensory neurons retrogradely labelled with FB were scattered in T5-L4 DRG but most were located at the upper lumbar levels (L1-L3); only a minor component of the extrinsic afferent innervation of the ileum was derived from nodose ganglia. In the DRG, 57% of retrogradely labelled neurons were also CGRP-IR. In cryostat sections, a dense network of CGRP-IR fibres was observed in the lamina propria beneath the epithelium, around the lacteals and lymphatic follicles (Peyer's platches), and along and around enteric blood vessels. Rare CGRP-IR fibres were also present in both muscle layers. Dense pericellular baskets of CGRP-IR fibres were observed around CGRP-negative somata. The only CGRP-IR nerve cells were well-defined Dogiel type II neurons localised in the MP and in the external and internal components of the SMP. CGRP-IR neurons in the myenteric ganglia were significantly larger than those in the submucosal ganglia (mean profile areas: about 1,400 μm2 for myenteric neurons, 750 μm2 for submucosal neurons). About 6% of myenteric neurons and 25% of submucosal neurons were CGRP-IR Dogiel type II neurons. The percentages of CGRP-IR neurons that were also tachykinin-IR were about 9% (MP) and 42% (SMP), whereas no CGRP-IR neurons exhibited immunoreactivity for vasoactive intestinal peptide, nitric oxide synthase or tyrosine hydroxylase in either plexus. Thus, CGRP immunoreactivity occurs in the enteric nervous system of the sheep ileum (as in human small intestine and MP of pig ileum) in only one morphologically defined type of neuron, Dogiel type II cells. These are probably intrinsic primary afferent neurons.

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Acknowledgements

We thank Valeria Albanese and Beatrice Travostini for their invaluable assistance and advice in the preparation of tissue for immunohistochemistry.

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Affiliations

  1. Department of Veterinary Morphophysiology and Animal Productions, University of Bologna, 40064, Bologna, Ozzano Emilia, Italy

    Roberto Chiocchetti, Annamaria Grandis, Cristiano Bombardi, Maria L. Lucchi, Davide Tadini Dal Lago & Ruggero Bortolami

  2. Department of Anatomy and Cell Biology & Centre for Neuroscience, University of Melbourne, Parkville, Melbourne, Australia

    John B. Furness

Authors
  1. Roberto Chiocchetti
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  2. Annamaria Grandis
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  3. Cristiano Bombardi
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  4. Maria L. Lucchi
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  5. Davide Tadini Dal Lago
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  6. Ruggero Bortolami
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  7. John B. Furness
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Corresponding author

Correspondence to Roberto Chiocchetti.

Additional information

This work was supported by grants from the Ricerca Fondamentale Orientata (RFO) and Fondazione Del Monte di Bo e Ra.

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Chiocchetti, R., Grandis, A., Bombardi, C. et al. Extrinsic and intrinsic sources of calcitonin gene-related peptide immunoreactivity in the lamb ileum: a morphometric and neurochemical investigation. Cell Tissue Res 323, 183–196 (2006). https://doi.org/10.1007/s00441-005-0075-2

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  • DOI: https://doi.org/10.1007/s00441-005-0075-2

Keywords

  • CGRP
  • Enteric nervous system
  • Dorsal root ganglion
  • Fluorescent tracer
  • Immunohistochemistry
  • Morphometry
  • Sheep