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Anatomical and neurochemical features of the extrinsic and intrinsic innervation of the striated muscle in the porcine esophagus: evidence for regional and species differences

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Abstract.

Studies of the intrinsic and extrinsic innervation patterns of esophageal motor endplates (MEPs) are mainly confined to small rodents. Therefore, an immunocytochemical, denervation and tracing study was conducted on the pig, an experimental model in which the distribution of the striated esophageal muscle portion more closely resembles the human situation. The purpose of this study was to analyze the origin and neurochemical content of the nerve fibers participating in the myoneural synapse. Fifteen 6-week-old domestic pigs were studied by immunohistochemistry combined with α-bungarotoxin labeling to define the co-innervation patterns of nitrergic and peptidergic nerve terminals in MEPs. Some animals were subjected to unilateral infra- or supranodose vagotomy to determine the origin of the nerve terminals in MEPs. Special attention was paid to the interregional differences in terms of co-innervation rates, and these findings were compared with literature data on small mammals. Double stainings revealed that most of the nNOS-immunoreactive (ir) terminals in MEPs co-stained for VIP, GAL and NPY, but not for PACAP and L-ENK. PACAP- and L-ENK-ir terminals were coarser than nNOS-ir terminals, and largely co-localized VAChT. High percentages of MEPs at the cervical level were contacted by PACAP- (~94%) and L-ENK-ir (~78%) terminals, but the proportion of both decreased in the rostrocaudal direction. Vagotomy significantly reduced their presence in MEPs at the thoracic and abdominal levels, while nNOS-ir terminals observed in approximately 30% of the MEPs were unaffected by vagotomy. Immunostainings on brainstem cryosections after retrograde tracing from the cervical esophagus showed that a large number of FB-positive cells in the nucleus ambiguus were PACAP-ir (~72%). C-kit-positive interstitial cells of Cajal were seen adjacent to the striated muscle fibers, apparently without direct relationship to MEPs. Similar to mouse esophagus, intrinsic nitrergic fibers were found to run close to, or even spiral around, these interstitial cells, an association that might point to a role as specialized spindle proprioceptors. In conclusion, the cholinergic terminals—part of which coexpress PACAP and/or L-ENK—that innervate MEPs in the porcine esophagus have a vagal origin, whereas the nNOS/VIP/GAL/NPY-ir fibers co-innervating these MEPs are intrinsic in nature. The regional differences observed along the esophageal length pertain to the neurochemical content of the vagal motor innervation of the MEPs.

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References

  • Biancani P, Harnett KM, Behar J (1999) Esophageal motor function. In: Yamada T (ed) Textbook of gastroenterology. JB Lippincott Co., Philadelphia, pp 157–187

  • Brouns I, Van Nassauw L, Van Genechten J, Majewski M, Scheuermann DW, Timmermans JP, Adriaensen D (2002) Triple immunofluorescence staining with antibodies raised in the same species to study the complex innervation pattern of intrapulmonary chemoreceptors. J Histochem Cytochem 50:575–582

    CAS  PubMed  Google Scholar 

  • Burns AJ, Herbert TM, Ward SM, Sanders KM (1997) Interstitial cells of Cajal in the guinea-pig gastrointestinal tract as revealed by c-kit immunohistochemistry. Cell Tissue Res 290:11–20

    CAS  PubMed  Google Scholar 

  • Calupca MA, Vizzard MA, Parsons RL (2000) Origin of pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive fibers innervating guinea-pig parasympathetic cardiac ganglia. J Comp Neurol 423:26–39

    CAS  PubMed  Google Scholar 

  • Collman PI, Tremblay L, Diamant NE (1993) The central vagal efferent supply to the esophagus and lower esophageal sphincter of the cat. Gastroenterology 104:1430–1438

    CAS  Google Scholar 

  • De Laet A, De Man JG, Cornelissen W, Schrödl F, Scheuermann DW, van Bogaert P-P, Neuhuber WL, Pelckmans PA, Timmermans J-P (1999) An electrophysiological and pharmacological study of the innervation of the striated musculature of the rat oesophagus. Neurogastroenterol Mot 11:257

    Google Scholar 

  • De Laet A, Cornelissen W, Adriaensen D, van Bogaert P-P, Scheuermann DW, Timmermans J-P (2002) Ca2+ involvement in the action potential generation of myenteric neurones in the rat oesophagus. Neurogastroenterol Mot 14:161–172

    Article  Google Scholar 

  • Félix B, Légeb M-E, Albe-Fessard D (1999) Stereotaxic atlas of the pig brain, 1st edn. Elsevier Science, The Netherlands

  • Fox EA, Phillips RJ, Byerly MS, Baronowsky EA, Chi MM, Powley TL (2002) Selective loss of vagal intramuscular mechanoreceptors in mice mutant for steel factor, the c-Kit receptor ligand. Anat Embryol 205:325–342

    PubMed  Google Scholar 

  • Geboes K, Desmet V (1978) Histology of the esophagus. Front Gastrointest Res 3:1–17

    CAS  PubMed  Google Scholar 

  • Hannibal J, Ekblad E, Mulder H, Sundler F, Fahrenkrug J (1998) Pituitary adenylate cyclase activating polypeptide (PACAP) in the gastrointestinal tract of the rat: distribution and effects of capsaicin or denervation. Cell Tissue Res 291:65–79

    CAS  PubMed  Google Scholar 

  • Hayakawa T, Takanaga A, Tanaka K, Maeda S, Seki M (2002) Organization and distribution of the upper and lower esophageal motoneurons in the medulla and the spinal cord of the rat. Okajimas Folia Anat Jpn 78:263–279

    PubMed  Google Scholar 

  • Kuramoto H, Endo Y (1995) Galanin-immunoreactive nerve terminals innervating the striated muscle fibers of the rat esophagus. Neurosci Lett 188:171–174

    CAS  PubMed  Google Scholar 

  • Kuramoto H, Kato Y, Sakamoto H, Endo Y (1996) Galanin-containing nerve terminals that are involved in a dual innervation of the striated muscles of the rat esophagus. Brain Res 734:186–192

    CAS  PubMed  Google Scholar 

  • Kuramoto H, Kawano H, Sakamoto H, Furness JB (1999) Motor innervation by enteric nerve fibers containing both nitric oxide synthase and galanin immunoreactivities in the striated muscle of the rat esophagus. Cell Tissue Res 295:241–245

    CAS  PubMed  Google Scholar 

  • Legradi G, Shioda S, Arimura A (1994) Pituitary adenylate cyclase-activating polypeptide-like immunoreactivity in autonomic regulatory areas of the rat medulla oblongate. Neurosci Lett 176:193–196

    CAS  PubMed  Google Scholar 

  • Morikawa S, Komuro T (1998) Distribution of myenteric NO neurons along the guinea-pig esophagus. J Autonom Nerv Syst 74:91–99

    Article  CAS  Google Scholar 

  • Neuhuber WL, Wörl J, Berthoud H-R, Conte B (1994) NADPH-diaphorase-positive nerve fibers associated with motor endplates in the rat esophagus—new evidence for co-innervation of striated muscle by enteric neurons. Cell Tissue Res 276:23–30

    CAS  PubMed  Google Scholar 

  • Neuhuber WL, Eichhorn U, Wörl J (2001) Enteric co-innervation of striated muscle fibers in the esophagus: just a 'hangover'? Anat Rec 262:41–46

    CAS  PubMed  Google Scholar 

  • Patapoutian A, Wold BJ, Wagner RA (1995) Evidence for developmentally programmed transdifferentiation in mouse esophageal muscle. Science 270:1818–1821

    CAS  PubMed  Google Scholar 

  • Reichel B, Wörl J, Neuhuber WL (1995) NPY-positive nerve fibers on motor endplates in the rat oesophagus stain also for VIP and NADPH-diaphorase. J Anat 187:226A

    Google Scholar 

  • Rodrigo J, Polak JM, Fernandez L, Ghatei MA, Mulderry P, Bloom SR (1985) Calcitonin gene-related peptide immunoreactive sensory and motor nerves of the rat, cat, and monkey esophagus. Gastroenterology 88:444–451

    CAS  PubMed  Google Scholar 

  • Rodrigo J, Pedrosa JA, Alvarez FJ, Bentura ML, Uttenthal O, Martinez-Murillo R, Polak JM (1994) Presence of calcitonin gene-related peptide in intraepithelial nerve fibers and motor end-plates of the cat esophagus: a light and electron microscopic study. J Auton Nerv Syst 49:21–31

    Google Scholar 

  • Rodrigo J, Uttenthal LO, Peinado MA, Esteban FJ, Fernandez AP, Serrano J, deVelasco JM, Santacana M, Bentura ML, MartinezMurillo R, Pedrosa JA (1998) Distribution of nitric oxide synthase in the esophagus of the cat and monkey. J Auton Nerv Syst 70:164–179

    CAS  PubMed  Google Scholar 

  • Rumessen JJ, de Kerchove d'Exaerde A, Mignon S, Bernex F, Timmermans J-P, Schiffmann SN, Panthier JJ, Vanderwinden JM (2001) Interstitial cells of Cajal in the striated musculature of the mouse esophagus. Cell Tissue Res 306:1–14

    CAS  PubMed  Google Scholar 

  • Sang Q, Young HM (1997) Development of nicotinic receptor clusters and innervation accompanying the change in muscle phenotype in the mouse esophagus. J Comp Neurol 386:119–136

    CAS  PubMed  Google Scholar 

  • Sang Q, Young HM (1998) The origin and development of the vagal and spinal innervation of the external muscle of the mouse esophagus. Brain Res 809:253–268

    CAS  PubMed  Google Scholar 

  • Sang Q, Ciampoli D, Greferath U, Sommer L, Young HM (1999) Innervation of the esophagus in mice that lack MASH1. J Comp Neurol 408:1–10

    CAS  PubMed  Google Scholar 

  • Sörensen B, Neuhuber WL, Mayer B, Wörl J (1995) Nitrerge Co-innervation motorischer Endplatten im Ösophagus bei Mensch und Schwein. Ann Anat 177:214A

    Google Scholar 

  • Storr M, Geisler F, Neuhuber WL, Allesher HD (1999) Autonomic modulation of vagal input to the rat esophageal muscle influence of opiates. Falk Symposium No. 112 Poster abstract I-4

  • Storr M, Geisler F, Neuhuber WL, Schusdziarra V, Allescher HD (2000) Endomorphin-1 and -2, endogenous ligands for the µ-opioid receptor, inhibit striated and smooth muscle contraction in the rat oesophagus. Neurogastroenterol Mot 12:441–448

    CAS  PubMed  Google Scholar 

  • Storr M, Geisler F, Neuhuber WL, Schusdziarra V, Allescher HD (2001) Characterization of vagal input to the rat esophageal muscle. Auton Neurosci Basic Clin 91:1–9

    Article  CAS  Google Scholar 

  • Thuneberg L (1999) One hundred years of interstitial cells of Cajal. Microsc Res Techn 47:223–238

    Article  CAS  Google Scholar 

  • Thuneberg L, Peters S (2001) Toward a concept of stretch-coupling in smooth muscle. I. Anatomy of intestinal segmentation and sleeve contractions. Anat Rec 262:110–124

    Article  CAS  PubMed  Google Scholar 

  • Timmermans J-P (2001) Interstitial cells of Cajal: is their role in gastrointestinal function in view of therapeutic perspectives underestimated or exaggerated? Folia Morphol 60:1–9

    CAS  Google Scholar 

  • Timmermans J-P, Scheuermann DW, Stach W, Adriaensen D, De Groodt-Lasseel MHA, Polak JM (1989) Neuromedin U-immunoreactivity in the nervous system of the small intestine of the pig and its coexistence with substance P and CGRP. Cell Tissue Res 258:331–337

    CAS  PubMed  Google Scholar 

  • Timmermans J-P, Barbiers M, Adriaensen D, Cornelissen W, Schemann M, Scheuermann DW (1996) Neurochemical identification of cholinergic and nitrergic neurons in pig small intestine. Ann Anat 178:59–60

    Google Scholar 

  • Wörl J, Mayer B, Neuhuber WL (1994) Nitrergic innervation of the rat esophagus: focus on motor endplates. J Auton Nerv Syst 49:227–233

    PubMed  Google Scholar 

  • Wörl J, Mayer B, Neuhuber WL (1997) Spatial relationships of enteric nerve fibers to vagal motor terminals and the sarcolemma in motor endplates of the rat esophagus: a confocal laser scanning and electron-microscopic study. Cell Tissue Res 287:113–118

    PubMed  Google Scholar 

  • Wörl J, Fischer J, Neuhuber WL (1998) Nonvagal origin of galanin-containing nerve terminals innervating striated muscle fibers of the rat esophagus. Cell Tissue Res 292:453–461

    PubMed  Google Scholar 

  • Wörl J, Dütsch F, Neuhuber WL (2002) Development of neuromuscular junctions in the mouse esophagus: focus on establishment and reduction of enteric co-innervation. Anat Embryol 205:141–152

    PubMed  Google Scholar 

  • Wu M, Timmermans J-P, Scheuermann DW (1997) Cholinergic, nitrergic and peptidergic innervation of the porcine esophagus. Ann Anat 179:342–343

    Google Scholar 

  • Zagorodnyuk VP, Brookes SJH (2000) Transduction sites of vagal mechanoreceptors in the guinea pig esophagus. J Neurosci 20:6249–6255

    CAS  PubMed  Google Scholar 

  • Zhou DS, Desaki J, Komuro T (1996) Neuro-muscular junctions of longitudinal and circular muscle fibers of the guinea-pig esophagus and their relation to myenteric plexus. J Auton Nerv Syst 58:63–68

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Cell Biology and Histology, Department of Biomedical Sciences, University of Antwerp (RUCA), Groenenborgerlaan 171, 2020, Antwerp, Belgium

    M. Wu, D. Adriaensen & J.-P. Timmermans

  2. Laboratoire de Neurophysiologie, Faculté de Medicine, Université Libre de Bruxelles, Brussels, Belgium

    J.-M. Vanderwinden

  3. Department of Clinical Physiology, University of Warmia and Mazury, Olsztyn, Poland

    M. Majewski & J. Wojtkiewicz

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  1. M. Wu
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  2. M. Majewski
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  3. J. Wojtkiewicz
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  4. J.-M. Vanderwinden
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  5. D. Adriaensen
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  6. J.-P. Timmermans
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Correspondence to J.-P. Timmermans.

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Wu, M., Majewski, M., Wojtkiewicz, J. et al. Anatomical and neurochemical features of the extrinsic and intrinsic innervation of the striated muscle in the porcine esophagus: evidence for regional and species differences. Cell Tissue Res 311, 289–297 (2003). https://doi.org/10.1007/s00441-002-0696-7

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