Summary
By use of the indirect immunofluorescence technique the distribution of calcitonin gene-related peptide (CGRP)-like immunoreactivity (LI) has been analyzed in cervical and lumbar dorsal root ganglia of untreated and colchicine-treated rats. In addition, lumbar ganglia were examined 2 weeks after transection of the sciatic nerve. The occurrence of CGRP-positive cells in relation to ganglion cells containing substance P-, somatostatin-, galanin-, cholecystokinin (CCK)-, and vasoactive intestinal polypeptide (VIP)/peptide histidine isoleucin (PHI)-LI has been evaluated on consecutive sections as well as using elution-restaining and double-staining techniques.
CGRP-LI was observed in many ganglion cells of all sizes ranging in diameter from 15 μm to 65 μm. Thus, this peptide occurs also in the large primary sensory neurons. In contrast to the sensory peptides described to date, CGRP-positive cells constituted up to 50% of all and 70% of the medium-sized neurons, thus being the most frequently occurring peptide in sensory neurons so far encountered. Subpulations of CGRP-positive neurons were shown to contain substance P-, somatostatin-, or galanin-LI and some CGRP-positive neurons contained both substance P- and galanin-LI. In fact, most substance P-, somatostatin- and galanin-positive cell bodies were CGRP-immunoreactive. The coexistence analysis further revealed that galanin and substance P often coexisted and that some cells contained both substance P- and somatostatin-LI, whereas no coexistence between galanin and somatostatin has as yet been seen. VIP/PHI-LI was only shown in a few cells in untreated or colchicine-treated rats. However, after transcetion of the sciatic nerve numerous VIP/PHI-positive cells were observed, some of which also contained CGRP-LI.
The present results indicate that a CGRP-like peptide is present in a wide range of primary sensory neurons probably not related to specific sensory modalities. Often this peptide coexists with other biologically active peptides. Taken together these findings suggest that CGRP may have a generalized function.
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References
Ainsworth A, Hall P, Wall PD, Allt G, MacKenzie ML. Gibson S, Polak JM (1981) Effects of capsaicin applied locally to adult peripheral nerve. II. Anatomy and enzyme and peptide chemistry of peripheral nerve and spinal cord. Pain 11:379–388
Andres KH (1961) Untersuchung über den Feinbau von Spinalganglien. Z Zellforsch 55:1–48
Amara SG, Jonas V, Rosenfeld MG, Ong ES, Evans RM (1982) Alternative RNA-processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature(London) 298:240–244
Arvidsson J, Ygge J, Grant G (1986) Cell loss in lumbar dorsal root ganglia and transganglionic degeneration after sciatic nerve resection in the rat. Brain Res 373:15–21
Basbaum AI, Glazer EJ (1983) Immunoreactive vasoactive intestinal polypeptide is concentrated in the sacral spinal cord: a possible marker for pelvic afferent fibers. Somatosens Res 1:69–82
Basbaum AI, Cruz L, Weber E (1986) Immunoreactive dynorphin B in primary afferent fibers of the cat. J Neurosci 6:127–133
Botticelli LJ, Cox BM, Goldstein A (1982) Immunoreactive dynorphin in mammalian spinal cord and dorsal root ganglia. Proc Nat Acad Sci USA 78:7783–7786
Bray D (1970) Surface movements during the growth of single explanted neurons. Proc Natl Acad Sci USA 65:905–910
Brodin E, Lindefors N, Dalsgaard C-J, Theodorsson-Norheim E, Rosell S (1986) Tachykinin mulitplicity in rat central nervous system as studied using antisera raised against substance P and neurokinin A. Regul Pept 13:253–272
Buchan AMJ, Sikora LKJ, Levy JG, McIntosh CHS, Dyck I, Brown JC (1985) An immunocytochemical investigation with monoclonal antibodies to somatostatin. Histochemistry 83:175–180
Charnay Y, Chayvialle J-A, Said SE, Dubois PM (1985) Localization of vasoactive intestinal peptide immunoreactivity in human foetus and newborn infant spinal cord. Neuroscience 14:195–205
Ch'ng JLC, Christofides ND, Anand P, Gibson SJ, Allen YS, Su HC, Tatemoto K, Morrison JFB, Polak JM, Bloom SR (1985) Distribution of galanin immunoreactivity in the central nervous system and the responses of galanin-containing neuronal pathways to injury. Neuroscience 16:343–354
Conrath-Verrier M, Dietl M, Tramu G (1984) Cholecystokinin-like immunoreactivity in the dorsal horn of the spinal cord of the rat: a light and electron microscopic study. Neuroscience 13:871–885
Coons AH (1958) Fluorescent antibody methods. In: Danielli JF (ed) General Cytochemical methods. Academic Press, New York, pp 399–422
Cuello AC, Del Fiacco M, Paxinos G (1978) The central and peripheral ends of the substance P-containing sensory neurons in the rat trigeminal system. Brain Res 152:499–510
Dahlström, A (1968) Effect of colchicine on amine storage granules in sympathetic nerves of rat. Eur J Pharmacol 5:111–113
Dalsgaard CJ, Vincent SR, Hökfelt T, Lundberg JM, Dahlström A, Schultzberg M, Dockray GJ, Cuello AC (1982) Coexistence of cholecystokininand substance P-like peptides in neurons of the dorsal root ganglia of the rat. Neurosci Lett 33:159–163
Dockray GJ, Williams RG, Zhu W-Y (1981) Development of region-specific antisera for the C-terminal tetrapeptide of gastrin/ cholecystokinin and their use in studies of immunoreactive forms of cholecystokinin in rat brain. Neurochem Int 3:281–288
Dodd J, Jahr CE, Jessell TM (1984) Neurotransmitters and neuronal markers at sensory synapses in the dorsal horn. In: Kruger L, Liebeskind JC (eds) Advances in pain research and therapy, Vol. 6. Raven Press, New York, pp 105–121
Duce IR, Keen P (1977) An ultrastructural classification of the neuronal cell bodies of the rat dorsal root ganglion using zinc iodide-osmium impregnation. Cell Tissue Res 185:263–277
Fahrenkrug J, Pedersen JH (1984) Development and validation of a specific radioimmunoassay for PHI in plasma. Clin Chim Acta 143:183–192
Fahrenkrug J, Schaffalitzky de Muckadell OB (1977) Radioimmunoassay of vasoactive intestinal polypeptide (VIP) in plasma. J Lab Clin Med 89:1379–1388
Fahrenkrug J, Schaffalitzky de Muckadell OB (1978) Distribution of vasoactive intestinal polypeptide (VIP) in the porcine central nervous system. J Neurochem 31:1445–1451
Frey P (1985) Cholecystokinin octapeptide (CCK 26–33), nonsulfated octapeptide and tetrapeptide (CCK 30–33) in rat brain: analysis by high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA). Neurochem Int 5:811–815
Fuji K, Senba E, Fujii S, Nomura I, Wu J-Y, Ueda Y, Tohyama M (1985) Distribution, ontogeny and projections of cholecystokinin-8, vasoactive intestinal polypeptide and γ-aminobutyrate-containing neuron systems in the rat spinal cord: an immunohistochemical analysis. Neuroscience 14:881–894
Gibbins IL, Furness JB, Costa M, MacIntyre I, Hillyard CJ, Girgis S (1985) Co-localization of calcitonin gene-related peptide-like immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs. Neurosci Lett 57:125–130
Gibson JG, Polak JM, Bloom SR, Wall PD (1981) The distribution of nine peptides in rat spinal cord with special emphasis on the substantia gelatinosa and on the area around the central canal (lamina X). J Comp Neurol 201:65–79
Gibson SJ, McGregor G, Bloom SR, Polak JM, Wall PD (1982) Local application of capsaicin to one sciatic nerve of the adult rat induces a marked depletion in the peptide content of the lumbar dorsal horn. Neuroscience 7:3153–3162
Gibson SJ, Polak JM, Anand P, Blank MA, Morrison JFB, Kelly JS, Bloom SR (1984a) The distribution and origin of VIP in the spinal cord of six mammalian species. Peptides 5:201–207
Gibson SJ, Polk JM, Bloom SR, Sabate IM, Mulderry PM, Ghatei MA, Morrison JFB, Kelly JS, Rosenfeld MG (1984b) Calcitonin gene-related peptide (CGRP)-immunoreactivity in the spinal cord of man and of eight species. J Neurosci 4:3101–3111
Gibson SJ, McCrossan MV, Polak JM (1985) A sub-population of calcitonin gene-related peptide (CGRP)-immunoreactive neurons in the dorsal root ganglia also display substance P, somatostatin or galanin immunoreactivity. In: Proc. XII International Anatomical Congress, Abstract A 232
Hanko J, Hardebo JE, Kåhrström J, Owman C, Sundler F (1985) Calcitonin gene-related peptide is present in mammalian cerebrovascular nerve fibres and dilates pial and peripheral arteries. Neurosci Lett 57:91–95
Hökfelt T, Johansson O, Luft R, Arimura A (1975b) Immunohistochemical evidence for the presence of somatostatin, a powerful inhibitory peptide, in some primary sensory neurons. Neurosci Lett 1:231–235
Hökfelt T, Elde R, Johansson O, Luft R, Nilsson G, Arimura A (1976) Immunohistochemical evidence for separate populations of somatostatin and substance P-containing primary neurons in the rat. Neuroscience 1:131–136
Hökfelt T, Johansson O, Ljungdahl Å, Lundberg JM, Schultzberg M (1980) Peptidergic neurons. Nature London 284:515–521
Hökfelt T, Schultzberg M, Lundberg JM, Fuxe K, Mutt V, Fahrenkrug J, Said SI (1982) Distribution of vasoactive intestinal polypeptide in the central and peripheral nervous systems as revealed by immunocytochemistry. In: Said SI (ed) Vasoactive intestinal peptide. Raven Press, New York, pp 65–90
Hökfelt T, Vincent S, Dalsgaard CJ, Skirboll L, Johansson O, Schultzberg M, Lundberg J, Rosell S, Pernow B, Janscó G (1982) Distribution of substance P in brain and periphery and its possible role as a co-transmitter. In: Porter R, O'Connor M (Eds) Substance P in the nervous system, Ciba Foundation Symposium, Vol 91. Pitman, London, pp 84–106
Hökfelt T, Skirboll L, Everitt BJ, Meister B, Brownstein M, Jacobs T, Faden A, Kuga S, Goldstein M, Markstein R, Dockray G, Rehfeld J (1985) Distribution of cholecystokinin-like immunoreactivity in the nervous system with special reference to coexistence with classical neurotransmitters and other neuropeptides. In: Vanderhaeghen JJ, Crawley J (eds) Neuronal cholecystokinin. Ann NY Acad Sci, New York, pp 255–274
Honda CN, Rethelyi M, Petrusz P (1983) Preferential immunohistochemical localization of vasoactive intestinal polypeptide (VIP) in the sacral spinal cord of the cat: light and electron microscopic observations. J Neurosci 3:2183–2196
Hyldén JLK, Wilcox GL (1981) Intrathecal substance P elicits a caudally directed biting and scratching behaviour in mice. Brain Res 217:212–215
Jacobs JM, Carmichael N, Cavanagh JB (1975) Ultrastructural changes in the dorsal root and trigeminal ganglia of rats poisoned with methyl mercury. Neuropathol Appl Neurobiol 1:1–19
Jancsó G, Hökelt T, Lundberg JM, Kiraly E, Halasz N, Nilsson G, Terenius L, Rehfeld J, Steinbusch H, Verhofstad A, Elde R, Said S, Brown M (1981) Immunohistochemical studies on the effect of capsaicin on peptide and monoamine neurons using antisera to substance P, gastrin/CCK, somatostatin, VIP, enkephalin, neurotensin and 5-hydroxytryptamine. J Neurocytol 10:963–980
Johnson DG, de C Nogueira Araujo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Meth 43:349
Ju G, Hökfelt T, Fischer JA, Frey P, Rehfeld JF, Dockray GJ (1986) Does cholecystokinin-like immunoreactivity in primary sensory neurons represent calcitonin gene-related peptide? Neurosci Lett 68:305–310
Kawatani M, Lowe IP, Nedelhaft I, Morgan C, de Groat WC (1983) Vasoactive intestinal polypeptide in the visceral afferent pathways to the sacral spinal cord of the cat. Neurosci Lett 42:311–316
Kessler JA, Adler JE, Bohn MC, Black IB (1981) Substance P in principal sympathetic neurons: regulation by impulse activity. Science 214:335–336
Knyihar-Csillik E, Csillik B (1981) FRAP: histochemistry of the primary nociceptive neuron. Prog Histochem Cytochem 14:1–137
Kuwayama Y, Stone RA (1986) Neuropeptide immunoreactivity of pericellular baskets in the guinea pig trigeminal ganglion. Neurosci Lett 64:169–172
Larsson L-I, Rehfeld JF (1979) Localization and molecular heterogeneity of cholecystokinin in the central and peripheral nervous system. Brain Res 165:201–218
Leah JD, Cameron AA, Kelly WL, Snow PJ (1985) Coexistence of peptide immunoreactivity in sensory neurons of the cat. Neuroscience 16:683–690
Lee Y, Takami K, Kawai Y, Girgis S, Hillyard CJ, MacIntyre I, Emson PC (1985a) Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P. Neuroscience 15:1227–1237
Lee Y, Kawai Y, Shiosaka S, Takami K, Kiyama H, Hillyard CJ, Girgis S, MacIntyre I, Emson PC, Tohyama M (1985b) Coexistence of calcitonin gene-related peptide and substance Plike peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis. Brain Res. 330:194–196
Le Greves P, Nyberg F, Terenius L, Hökfelt T (1985) Calcitonin gene-related peptide is a potent inhibitor of substance P degradation. Eur J Pharmacol 115:309–311
Lembeck F (1953) Zur Frage der zentralen Übertragung afferenter Impulse. III. Mitteilung. Das Vorkommen und die Bedeutung der Substanz P in den dorsalen Wurzeln des Rückenmarks. Naunyn-Schmiedeberg's Arch Pharmacol 219:197–213
Liebermann AR (1976) Sensory ganglia. In: Landon DN (ed) The peripheral nerve. Chapman and Hall, London, pp 188–278
Lorén I, Alumets J, Håkanson RH, Sundler F (1979) Distribution of gastrin and cholecystokinin-like peptides in rat brain. Histochemistry 59:249–257
Lundberg JM, Hökfelt T, Nilsson G, Terenius L, Rehfeld JF, Elde R, Said S (1978) Peptide neurons in the vagus, splanchnic and sciatic nerves. Acta Physiol Scand 104:499–501
Lundberg JM, Franco-Cereceda A, Hua X, Hökfelt T, Fischer JA (1985) Co-existence of substance P and calcitonin gene-related peptide-like immunoreactivities in sensory nerves in relation to cardiovascular and bronchoconstrictor effects of capsaicin. Eur J Pharmacol 108:315–319
Maderdrut JL, Yaksh TL, Petrusz P, Go VLW (1982) Origin and distribution of cholecystokinin-containing nerve terminals in the lumbar dorsal horn and nucleus caudalis of the cat. Brain Res 243:363–368
Marley PD, Nagy JE, Emson PC, Rehfeld JF (1982) Cholecystokinin in the rat spinal cord: distribution and lack of effect of neonatal capsaicin treatment and rhizotomy. Brain Res 238:494–498
Matsuyama T, Wanaka A, Yoneda S, Kimura K, Kamada T, Girgis S, MacIntyre I, Emson PC, Tohyama M (1986) Two distinct calcitonin gene-related peptide-containing peripheral nervous systems: distribution and quantitative difference between the iris and cerebral artery with special reference to substance P. Brain Res 373:205–212
McGregor GP, Gibson SJ, Sabate IM, Blank MA, Christofides ND, Wall PD, Polak JM, Bloom SR (1984) Effect of peripheral nerve section and nerve crush on spinal cord neuropeptides in the rat: increased VIP and PHI in the dorsal horn. Neuroscience 13:207–216
Morgan C, de Groat WC, Jannetta PJ (1982) Leucine enkephalin and substance-P identified in trigeminal ganglion neurons innervating the cornea in the cat. Neuroscience [Abstr] 8:474
Mulderry PK, Ghatel 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
Nagy JI, Hunt SP (1982) Fluoride-resistant acid phosphatase-containing neurones in dorsal root ganglia are separate from those containing substance P or somatostatin. Neuroscience 7:89–97
Nagy JI, Goedert H, Hunt SP, Bond A (1982) The nature of the substance P-containing nerve fibres in taste papillae of the rat tongue. Neuroscience 73137–3151
Nairn RC (1969) Immunological tracing: general considerations. In: Nairn RC (ed) Fluorescent protein tracing. Livingstone LTD, Edinburgh London, pp 111–151
Otten U, Lorez HP (1982) Nerve growth factor increases substance P, cholecystokinin, and vasoactive intestinal polypeptide immunoreactivities in primary sensory neurones of newborn rats. Neurosci Lett 34:153–158
Pernow B (1953) Studies on substance P. Purification, occurrence and biological actions. Acta Physiol Scand 105 [Suppl] 29:1–90
Piercy MF, Dobry PJK, Schroeder LA, Einsphar FJ (1981) Behavioral evidence that substance P may be a spinal cord sensory neurotransmitter. Brain Res 210:407–412
Price J, Mudge AW (1983) A subpopulation of rat dorsal root ganglion neurons are catecholaminergic. Nature 301:241–243
Price J (1985) An immunohistochemical and quantitative examination of dorsal root ganglion neuronal sub-populations. J Neurosci 5:2051–2059
Rambourg A, Clemont Y, Beaudet A (1983) Ultrastructural features of six types of neurones in rat dorsal root ganglia. J Neurocytol 12:47–66
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
Rosenfeld MG, Mermod JJ, Amara SG, Swanson LW, Sawchenko PE, Rivier J, Vale WW, Evans RH (1983) Production of a novel neuropeptide encoded by the calcitonin gene via tissue specific RNA processing. Nature (London) 304:129–135
Schultzberg M, Williams RG, Dockray GJ (1981) Capsaicin depletes substance P, but not CCK-8 in dorsal spinal cord. Neurosci Lett [Suppl] 7:S321
Schultzberg M, Dockray GJ, Williams RG (1982) Capsaicin depletes CCK-like immunoreactivity detected by immunohistochemistry, but not that measured by radioimmunoassay in rat dorsal spinal cord. Brain Res 235:198–204
Senba E, Shiosaka S, Hara Y, Inagaki S, Sakanaka M, Takatsuki K, Kawai Y, Tohyama M (1982) Ontogeny of the peptidergic system in the rat spinal cord: immunohistochemical analysis. J Comp Neurol 208:54–66
Shehab SAS, Atkinson ME (1984) Sciatic nerve section has variable effects on primary afferent neuropeptides. J Anat (Lond) 139:725
Shehab SAS, Atkinson (1986) Vasoactive intestinal polypeptide increases in areas of the dorsal horn of the spinal cord from which other neuropeptides are depleted following peripheral axotomy. Exp Brain Res 62:422–430
Skofitsch G, Zamir N, Helke CJ, Savin JM, Jacobowitz DM (1985) Corticotropin releasing factor-like immunoreactivity in sensory ganglia and capsaicin sensitive neurons of the rat central nervous system: colocalization with other neuropeptides. Peptides 6:307–318
Skofitsch G, Jacobowitz DM (1985b) Calcitonin gene-related peptide coexists with substance P in capsaicin sensitive neurons and sensory ganglia of the rat. Peptides 6:747–754
Skofitsch G, Jacobowitz DM (1985c) Galanin-like immunoreactivity in capsaicin sensitive sensory neurons and ganglia. Brain Res Bull 15:191–195
Sweetnam PM, Neale JH, Barker JL, Goldstein A (1982) Localization of immunorcactive dynorphin in neurons cultured from spinal cord and dorsal root ganglia. Proc Natl Acad Sci USA 79:6742–6746
Takahashi T, Konishi S, Powcll D, Leeman SE, Otsuka M (1974) Identification of the motoneuron-depolarizing peptide in bovine dorsal root as hypothalamic substance P. Brain Res 73:59–69
Terenghi G, Polak JM, Ghatei MA, Mulderry PK, Butler JM, Unger WG, Bloom SR (1985) Distribution and origin of calcitonin gene-related peptide (CGRP) immunoreactiviy in the sensory innervation of the mammalian eye. J Comp Neurol 233:506–516
Terenghi G, Polak JM, Rodrigo J, Mulderry PK, Bloom SR (1986) Calcitonin gene-related peptide-immunorcactive nerves in the tongue, epiglottis and pharynx of the rat: occurrence, distribution and origin. Brain Res 365:1–14
Tramu G, Pillez A, Leonardelli J (1978) An efficient method of antibody elution for the successive or simultaneous location of two antigens by immunocytochemistry. J Histochem Cytochem 26:322–324
Tschopp FA, Tobler PH, Fischer JA (1984) Calcitonin gene-related peptide in the human thyroid, pituitary and brain. Mol Cell Endocrinol 36:53–57
Tschopp FA, Henke H, Petermann JB, Tobler PH, Janzer R, Hökfelt T, Lundberg JM, Cuello C, Fischer JA (1985) Calcitonin gene-related peptide and its binding sites in the human central nervous system and pituitary. Proc Natl Acad Sci USA 82:248–252
Tuchscherer MM, Seybold VS (1985) Immunohistochemical studies of substance P, cholecystokinin-octapeptide and somatostatin in dorsal root ganglia of the rat. Neuroscience 14:593–605
Uddman R, Edvinsson L, Ekman R, Kingman T, McCulloch J (1985a) Innervation of the feline cerebral vasculature by nerve fibers containing calcitonin gene-related peptide: trigeminal origin and co-existence with substance P. Neurosci Lett 62:131–136
Uddman R, Luts A, Sundler F (1985b) Occurrence and distribution of calcitonin gene-related peptide in the mammalian respiratory tract and middle ear. Cell Tissue Res 241:551–555
Vanderhaeghen J-J, Lotstra F, De Mey J, Gilles C (1980) Immunohistochemical localization of cholecystokininand gastrin-like peptides in the brain and hypophysis of the rat. Proc Natl Acad Sci USA 77:1190–1194
Wanaka A, Matsuyama T, Yoneda S, Kimura K, Kamada T, Girgis S, MacIntyre I, Emson PC, Tohyama M (1986) Origins and distribution of calcitonin gene-related peptide-containing nerves in the wall of the cerebral arteries of the guinea pig with special reference to the coexistence with substance P. Brain Res 369:185–192
Weihe E, Hartschuh W, Weber E (1985) Prodynorphin opioid peptides in small somatosensory primary afferents of guinea pig. Neurosci Lett 58:347–352
Wiesenfeld-Hallin Z, Hökfelt T, Lundberg JM, Forssmann WG, Reinecke M, Tschopp FA, Fischer (1984) Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neurons to the spinal cord and interact in spinal behavioral responses of the rat. Neurosci Lett 52:199–204
Woolf C, Wiesenfeld-Hallin Z (1986) Substance P and calcitonin gene related peptide synergistically modulate the gain of the nociceptive flexor withdrawal reflex in the rat. Neurosci Lett (in press)
Yokokawa K, Tohyama M, Shiosaka S, Shiotani Y, Sonoda T, Emson PC, Hillyard CJ, Girgis S, MacIntyre I (1986) Distribution of calcitonin gene-related peptide-containing fibers in the urinary bladder of the rat and their origin. Cell Tissue Res 244:271–278
Zamboni L, DeMartino C (1967) Buffered picric-acid formaldehyde: a new rapid fixative for electron-microscopy. J Cell Biol 35:148A
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Ju, G., Hökfelt, T., Brodin, E. et al. Primary sensory neurons of the rat showing calcitonin gene-related peptide immunoreactivity and their relation to substance P-, somatostatin-, galanin-, vasoactive intestinal polypeptide- and cholecystokinin-immunoreactive ganglion cells. Cell Tissue Res. 247, 417–431 (1987). https://doi.org/10.1007/BF00218323
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DOI: https://doi.org/10.1007/BF00218323