Abstract
At present our knowledge of enteric peptide-containing neurons in man is limited. In this study we have used human appendices removed at surgery to examine the peptidergic innervation by immunocytochemistry, immunochemistry, and pharmacological in vitro experiments. Immunocytochemistry revealed a variety of peptide-containing nerve fiber populations in the human appendix. VIP/PHI-, VIP/PHI/NPY-, SP/NKA-, galanin-, and enkephalin-containing nerve fibers were numerous; CGRP- and GRP- containing nerve fibers were moderate in number, while only scattered NPY-, enkephalin/BAM-, and somatostatin-containing nerve fibers could be found. No CCK-, dynorphin A-, or dynorphin B- immunoreactive nerve fibers could be detected. The coexistence of VIP/PHI, SP/NKA, and enkaphalin/BAM can be anticipated from the known sequence of their respective precursors. However, the coexistence of VIP/PHI and NPY was unexpected but corroborates previous observations in other species. Interestingly, SP and CGRP did not seem to coexist in nerve fibers of the human appendix. Immunochemistry (RIA and HPLC) confirmed the presence of VIP, NPY, SP, galanin, CGRP, GRP, enkephalin, and somatostatin. Motor activity studies suggest that acetylcholine plays a major role in the electrically evoked contractions, since atropine suppressed these contractions. Galanin (10−8-10−6 M) and GRP (10−9-10−7 M) caused concentration-dependent contractions that were unaffected by tetrodotoxin and thus probably reflect a direct action on smooth muscle receptors. GRP (10−9 M) enhanced the electrically induced cholinergic contraction (to 193±24%), while met-enkephalin (10−6 M) reduced it (to 54±6%). Both peptides failed to affect the contractile response to exogenous acetylcholine and probably act to modulate the release of acetylcholine. NPY, VIP, CGRP, SP, and somatostatin failed to induce contraction or to affect the electrically evoked contractions.
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References
Hökfelt T, Johansson O, Ljungdahl å, Lundberg JM, Schultzberg M: Peptidergic neurones. Nature 284:515–521, 1980
Sundler F, Håkanson R, Leander S: Peptidergic nervous systems in the gut. Clin Gastroenterol 9:517–543, 1980
Costa M, Furness JB, Llewellyn-Smith IJ: Histochemistry of the enteric nervous system.In Physiology of the Gastrointestinal Tract, 2nd ed, LR Johnson (ed). New York, Raven Press, 1987, pp 1–31
Polak JM, Bloom SR: Distribution, origin, and pathology of the gut peptidergic innervation.In Cellular Basis of Chemical Messengers in the Digestive Tract. M Grossman, MAB Brazier, J Lechago (eds). New York, Academic Press, 1981, pp 267–282
Makhlouf GM: Enteric neuropeptides: Role in neuromuscular activity of the gut. Trends Pharmacol Sci 6:214–218, 1985
Ashley DJB: Observations on the epidemiology of appendicitis. Gut 8:533–538, 1967
Creed F: Life events and appendectomy. Lancet 1:1381–1385, 1981
Masson P: Neuronal proliferations in the vermiform appendix.In Cytology of the Nervous System, Vol III, W Penfield (ed). 1932, pp 1095–1130
Auböck L, Ratzenhofer M: “Extraepithelial enterochromaffin cell-nerve fibre complexes” in the normal human appendix, and in neurogenic appendicopathy. J Pathol 136:217–226, 1982
Höfler H, Kasper M, Heitz PU: The neuroendocrine system of normal human appendix, ileum and colon, and in neurogenic appendicopathy. Virchows Arch A (Pathol Anat) 399:127–140, 1983
Ekblad E, Håkanson R, Sundler F: VIP and PHI coexist with an NPY-like peptide in intramural neurones of the small intestine. Regul Pept 10:47–55, 1984
Yanaihara N, Yanaihara C, Mochizuki T, Imura K, Fujita T, Iwanaga T: Immunoreactive GRP. Peptides 2(Suppl 2): 185–192, 1981
Brodin E, Alumets J, Håkanson R, Leander S, Sundler F: Immunoreactive substance P in chicken gut: Distribution, development and possible functional significance. Cell Tissue Res 216:455–469, 1981
Brodin E, Lindefors N, Theodorsson-Norheim E, Peterson L, Bartfai T, ögren S-O, Rosell S: Tachykinins in rat central nervous system: Distribution, molecular forms, release and effects of chronic treatment with antidepressent drugs.In Tachykinin Antagonists, Proceedings, Fernström Symposium, Vol 6. R Håkanson, F Sundler (eds). Amsterdam, Elsevier, 1985, pp 3–14
Alumets J, Håkanson R, Sundler F, Chang KJ: Leuenkephalin-like material in nerves and enterochromaffin cells in the gut. Histochemistry 56:187–196, 1978
Ekblad E, Ekelund M, Graffner H, Håkanson R, Sundler F: Peptide containing nerve fibers in the stomach wall of rat and mouse. Gastroenterology 89:73–85, 1985
Sundler F, Moghimzadeh E, Håkanson R, Ekelund M, Emson P: Nerve fibers in the gut and pancreas of the rat displaying neuropeptide-Y immunoreactivity. Intrinsic and extrinsic origin. Cell Tissue Res 230:487–493, 1983
Ekblad E, Winther C, Ekman R, Håkanson R, Sundler F: Projections of peptide-containing neurons in rat small intestine. Neuroscience 20:169–188, 1987
Ekblad E, Rökaeus å, Håkanson R, Sundler F: Galanin nerve fibers in the rat gut: distribution, origin and projections. Neuroscience 16:355–363, 1985
Lorén I, Alumets J, Håkanson R, Sundler F: Distribution of gastrin- and CCK-like peptides in rat brain. An immunocytochemical study. Histochemistry 59:249–257, 1979
Sundler F, Brodin E, Ekblad E, Håkanson R, Uddman R: Sensory nerve fibers: Distribution of substance P, neurokinin A and calcitonin gene-related peptide.In Tachykinin Antagonists, Proceedings, Fernström Symposium, R Håkanson, F Sundler (eds). Amsterdam, Elsevier, 1985, pp 3–14
Coons AH, Leduc EH, Connolly JM: Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J Exp Med 102:49–60, 1955
Tramu G, Pillez A, Leonardelli J: An efficient method of antibody elution for the successive or simultaneous location of two antigens by immunocytochemistry. J Histochem Cytochem 26:322–324, 1978
Cuello AC, Milstein C, Couture R, Wright B, Priestley JV, Jarvis J: Characterization and immunocytochemical application of monoclonal antibodies against enkephalins. J Histochem Cytochem 32:947–957, 1984
Fahrenkrug J, Schaffalitzky de Muckadell OB: Radioimmunoassay of VIP in plasma. J Lab Clin Med 89:1379, 1977
Voigt KH, Weber E, Martin R: Neuropeptides, subcellular localization of ACTH and related peptides.In Structure and Activity of Natural Peptides. W Woelter, G Weitzel (eds). Berlin, de Gruyter, 1980, pp 115–140
Grunditz T, Ekman R, Håkanson R, Rerup C, Sundler F, Uddman R: Calcitonin gene-related peptide in thyroid nerve fibers and C cells: Effects on thyroid hormone secretion and response to hypercalcemia. Endocrinology 119:2313–2324, 1987
Ekman R, Widerlöv E, Walleus H, Lindström LH: Elevated levels of NPY-like material in CSF in depression. ISPN XVth International Congress, Vienna. July 15–19, 1984, p 73
Wallengren J, Ekman R, Sundler F: Occurrence and distribution of neuropeptides in the human skin. Acta Derm Venereol 67:185–192, 1987
Ishida Y, Krakawa N: Isometric and isotonic spontaneous contractions of guinea-pig taenia coli. Jpn J Pharmacol 24:925–927, 1974
Leander S, Håkanson R, Sundler F: Nerves containing substance P, vasoactive intestinal polypeptide, enkephalin or somatostatin in the guinea pig taenia coli. Cell Tissue Res 215:21–39, 1981
Bishop AE, Ferri GL, Probert L, Bloom SR, Polak JM: Peptidergic nerves. Scand J Gastroenterol (Suppl 71) 17:43–59, 1981
Ferri GL, Potti PL, Vezzadin P, Biliotti G, Bloom SR, Polak JM: Peptide-containing innervation of the human intestinal mucosa. Histochemistry 76:413–420, 1982
Larsson LT, Malmfors G, Sundler F: Peptidergic innervation in Hirschsprung's disease. Z Kinderchir 38:301–304, 1983
Sjölund K, Schaffalitzky de Muckadell OB, Fahrenkrug J, Håkanson R, Peterson BG, Sundler F: Peptide-containing nerve fibres in the gut wall in Crohn's disease. Gut 24:724–733, 1983
Taguchi Y, Tanaka K, Ikeda K, Matsubayashi S, Yanaihara N: Peptidergic innervation irregularities in Hirschsprung's disease. Virchows Arch A 401:223–235, 1983
Tsuto T, Okamura H, Fukui K, Obata-Tsuto HL, Terubayashi H, Yanaighara J, Iwai N, Majima S, Yanaihara N, Ibata Y: Immunohistochemical investigations of gut hormones in the colon of patients with Hirschsprung's disease. J Pediatr Surg 20:266–270, 1985
Keast JR, Furness JB, Costa M: Distribution of certain peptide-containing nerve fibers and endocrine cells in the gastrointestinal mucosa in five mammalian species. J Comp Neurol 236:403–422, 1985
Wattchow DA, Furness JB, Costa M: Distribution and coexistence of peptides in nerve fibers of the external muscle of the human gastrointestinal tract. Gastroenterology 95:32–41, 1988
Ito S, Iwanaga T, Yamada Y, Shibata A: Somatostatin-28 like immunoreactivity in the human gut. Horm Metab Res 14:500–501, 1982
Larsson LT, Malmfors G, Sundler F: Neuropeptide Y, calcitonin gene-related peptide and galanin in Hirschsprung's disease: An immunocytochemical study. J Pediatr Surg 23:342–345, 1988
Itoh N, Obata K, Yanaihara N, Okamoto H: Human preprovasoactive intestinal polypeptide contains a novel PHI-27-like peptide, PHM-27. Nature 304:547–549, 1983
Nawa H, Hirose T, Takashima H, Inayama S, Nakanishi S: Nucleotide sequences of cloned cDNAs for two types of bovine brain substance P precursor. Nature 306:31–36, 1983
Mizuno K, Minamino N, Kangawa K, Matsuo H: A new family of endogenous “big” met-enkephalins from bovine adrenal medulla: Purification and structure of docosa- (BAM-22P) and eicosapeptide (BAM-20P) with very potent opiate activity. Biochem Biophys Res Commun 97:1283–1290, 1980
Gibbins IL, Furness JB, Costa M, MacIntyre I, Hillyard CJ, Girgis S: Co-localization of calcitonin gene-related peptidelike immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs. Neurosci Lett 57:125–130, 1985
Burnstock G, Campell G, Rand MJ: The inhibitory innervation of the taenia of the guinea pig caecum. J Physiol 182:504–526, 1966
Leander S, Ekman R, Uddman R, Sundler F, Håkanson R: Neuronal cholecystokinin, gastrin-releasing peptide, neurotensin and Β-endorphin in the intestine of the guinea pig. Distribution and possible motor function. Cell Tissue Res 235:521–531, 1984
Bitar KN, Makhlouf GM: Specific opiate receptors on isolated mammalian gastric smooth muscle cells. Nature 297:72–74, 1982
Dockray G: Physiology of Enteric Neuropeptides.In Physiology of the Gastrointestinal Tract, 2nd ed. LR Johnson (ed). New York, Raven Press, 1987, pp 41–66
Bertaccini G: Peptides: Candidate hormones.In Handbook of Experimental Pharmacology. G Bertaccini (ed). Berlin, Springer-Verlag, 1982, pp 85–135
Ekblad E, Håkanson R, Sundler F, Wahlestedt C: Galanin: Neuromodulatory and direct contractile effects on smooth muscle preparations. Br J Pharmacol 86:241–246, 1985
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Ekblad, E., Arnbjörnsson, E., Ekman, R. et al. Neuropeptides in the human appendix. Digest Dis Sci 34, 1217–1230 (1989). https://doi.org/10.1007/BF01537270
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DOI: https://doi.org/10.1007/BF01537270