Summary
The ventral medulla oblongata of rats was analyzed with a double-labelling immunofiuorescence technique using guinea pig antibodies directed against serotonin (5-HT) and rabbit antisera directed against enkephalin (ENK). Numerous cells in the region of nucleus raphe obscurus, nucleus raphe pallidus and nucleus raphe magnus showed immuno-staining for either 5-HT or ENK. A substantial number of cells showed positive immunostaining for both 5-HT and ENK. 5-HT/ENK double-labelled cells were most frequently encountered in an area that extended from the rostral aspect of the inferior olivary nucleus to the pontomedullary border. This region corresponds anatomically to nucleus raphe magnus/nucleus paragigantocellularis. In addition, a number of the 5-HT/ENK-containing cells were retrogradely labelled with Fluoro-Gold dye that had been injected into the thoracic spinal cord several days prior to perfusion. Schematic drawings showing the anatomical distribution of 5-HT/ENK colocalization are provided.
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Armstrong DM, Miller RJ, Beaudet A, Pickel VM (1984) Enkephalin-like immunoreactivity in rat area postrema: ultrastructural localization and coexistence with serotonin. Brain Res 310: 269–278
Basbaum AI, Fields HL (1984) Endogenous pain control systems: brainstem spinal pathways and endorphin circuitry. Ann Rev Neurosci 7: 309–338
Bowker RM, Steinbusch HWM, Coulter JD (1981) Serotonergic and peptidergic projections to the spinal cord demonstrated by a combined retrograde HRP histochemical and immunocytochemical staining method. Brain Res 211: 412–416
Burnstock G (1976) Do some nerve cells release more than one transmitter? Neuroscience 1: 239–248
Carlsson A, Falck B, Fuxe K, Hillarp N (1964) Cellular localization of monoamines in the spinal cord. Acta Physiol Scand 60: 112–119
Caverson MM, Ciriello J, Calaresu FR (1983) Direct pathway from cardiovascular neurons in the ventrolateral medulla to the region of the intermediolateral nucleus of the upper thoracic cord: an anatomical and electrophysiological investigation in the cat. J Auton Nerv Syst 9: 451–475
Chan-Palay V, Jonsson G, Palay SL (1978) Serotonin and substance P coexist in neurons of the rat's central nervous system. Proc Natl Acad Sci USA 75: 1582–1586
Coons AH (1958) Fluorescent antibody methods. In: Danielli JF (ed) General cytochemical methods. Academic Press, New York, pp 399–422
Cuello AC (ed) (1982) Co-transmission. MacMillan, London
Dahlström A, Fuxe K (1965) Evidence for the existence of monoamine neurons in the central nervous system. II. Experimentally induced changes in the intraneuronal amine levels of the bulbospinal neuron systems. Acta Physiol Scand Suppl 247: 1–36
Dampney RAL, Goodchild AK, Robertson LG, Montgomery W (1982) Role of ventrolateral medulla in vasomotor regulation: a correlative anatomical and physiological study. Brain Res 249: 223–235
Fields HL, Basbaum AI, Clanton CH, Anderson SD (1977) Nucleus raphe magnus inhibition of spinal cord dorsal horn neurons. Brain Res 126: 441–453
Finley JCW, Maderdrut JL, Petrusz P (1981) The immunohistochemical localization of enkephalin in the central nervous system of the rat. J Comp Neurol 198: 541–565
Fuxe K (1965) Evidence for the existence of monoamine neurons in the central nervous system. IV. Distribution of monoamine nerve terminals in the central nervous system. Acta Physiol Scand 64, Suppl 247: 37–85
Glazer EJ, Steinbusch H, Verhofstad A, Basbaum AI (1981) Serotonin neurons in nucleus raphe dorsalis and paragigantocellularis of the cat contain enkephalin. J Physiol (Paris) 77: 241–245
Hunt SP, Lovick TA (1982) The distribution of serotonin, metenkephalin and B-lipoprotein-like immunoreactivity in neuronal perikarya of the cat brainstem. Neurosci Lett 30: 139–145
Hökfelt T, Elde R, Johansson O, Terenius L, Stein L (1977) The distribution of enkephalin-immunoreactive cell bodies in the rat central nervous system. Neurosci Lett 5: 25–31
Hökfelt T, Ljungdahl, Steinbusch H, Verhofstad A, Nilsson G, Brodin E, Pernow B, Goldstein M (1978) Immunohistochemical evidence of substance P-like immunoreactivity in some 5- hydroxytryptamine-containing neurons in the rat central nervous system. Neuroscience 3: 517–538
Hökfelt T, Millhorn D, Seroogy K, Tsuruo Y, Ceccatelli S, Lindh B, Meister B, Melander T, Schalling M, Terenius L (1987) Coexistence of peptides with classical neurotransmitters. Experientia 43: 768–779
Johansson O, Hökfelt T, Pernow B, Jeffcoate SL, White N, Steinbusch HWM, Verhofstad AAJ, Emerson PC, Spindel E (1981) Immunohistochemical support for three putative transmitters in one neuron: coexistence of 5-hydroxytryptamine-, substance P-, and thyrotropin releasing hormone-like immunoreactivity in medullary neurons projecting to the spinal cord. Neuroscience 6: 1857–1881
Johnson DG, De C Nogueira Aruajo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Meth 43: 349–350
Leger L, Charney Y, Dubois PM, Jouvet M (1986) Distribution of enkephalin-immunoreactive cell bodies in relation to serotonin-containing neurons in the raphe nuclei of the cat: immunohistochemical evidence for the coexistence of enkephalins and serotonin in certain cells. Brain Res 362: 63–73
Light AR, Casale EJ, Menetrey DM (1986) The effects of focal stimulation in nucleus raphe magnus and periaqueductal gray on intracellularly recorded neurons in spinal laminae I and II. J Neurophysiol 56: 555–571
Loewy AD, McKellar S (1981) Serotonergic projections from the ventral medulla to the intermediolateral cell column in the rat. Brain Res 211: 146–152
Melander T, Hökfelt T, Rökaeus A, Cuello AC, Oertel WH, Verhofstad A, Goldstein M (1987) Coexistence of galaninlike immunoreactivity with catecholamines, 5-hydroxytryptamine, GABA and neuropeptides in the rat CNS. J Neurosci 6: 3640–3654
Merchenthaler I, Maderdrut JL, Dockray GJ (1986) Immunocytochemical localization of proenkephalin-derived peptides in the central nervous system of the rat. Neuroscience 17: 325–348
Millhorn DE (1986) Neural respiratory and circulatory interaction during chemoreceptor stimulation and cooling of ventral medulla in cats. J Physiol (Lond) 370: 217–231
Millhorn DE, Eldridge FL (1986) Role of ventrolateral medulla in regulation of respiratory and cardiovascular systems. J Appl Physiol 61: 1249–1263
Millhorn DE, Hökfelt T (1988) Chemical messengers and their coexistence in individual neurons. News in Physiol Sci 3: 1–5
Millhorn DE, Hökfelt T, Seroogy K, Oertel W, Verhofstad AAJ, Wu J-Y (1987) Immunohistochemical evidence for colocalization of gamma-aminobutyric acid and serotonin in neurons of the ventral medulla oblongata projecting to the spinal cord. Brain Res 410: 179–185
Millhorn DE, Hökfelt T, Seroogy K, Verhofstad AAJ (1988) Extent of colocalization of serotonin and GABA in neurons of the ventral medulla oblongata in rat. Brain Res 461: 169–174
Millhorn DE, Hökfelt T, Terenius L, Buchan A, Brown JC (1987) Somatostatin-and enkephalin-like immunoreactivities are frequently colocalized in neurons in the caudal brain stem of rat. Exp Brain Res 67: 420–428
Oliveras JL, Guilbaud G, Besson JM (1979) A map of serotonergic structures involved in stimulation producing analgesia in unrestrained freely moving cats. Brain Res 164: 317–322
Petrusz P, Merchenthaler I, Maderdrut JL (1985) Distribution of enkephalin-containing neuron in the central nervous system. In: Björklund A, Hökfelt T (eds) Handbook of chemical neuroanatomy, Vol 4. GABA and neuropeptides in the CNS, Part 1. Elsevier, Amsterdam, pp 273–334
Platt JL, Michael AF (1983) Retardation of fading and enhancement of intensity of immunofluorescence by p-phenylenediamine. J Histochem Cytochem 31: 840–842
Rivot JP, Chaouch A, Besson JM (1980) Nucleus raphe magnus modulation of responses of rat dorsal horn neurons to unmyelinated fiber inputs: partial involvement of serotonergic pathways. J Neurophysiol 44: 1039–1057
Sar M, Stumpf WE, Miller RJ, Chang K-J, Cuatrecasas P (1978) Immunohistochemical localization of enkephalin in rat brain and spinal cord. J Comp Physiol 182: 17–38
Schmued LC, Fallon JH (1986) Fluoro-gold: a new fluorescent retrograde axonal tracer with numerous unique properties. Brain Res 377: 147–154
Schultzberg M, Lundberg JM, Hökfelt T, Terenius L, Brandt J, Elde RP, Goldstein M (1978) Enkephalin-like immunoreactivity in gland cells and nerve terminals of the adrenal medulla. Neuroscience 3: 1169–1186
Steinbusch HWM (1981) Distribution of serotonin-immunoreactivity in the central nervous system of the rat-cell bodies and terminals. Neuroscience 6: 557–618
Uhl GR, Goodman RR, Kuhar MJ, Childers SR, Snyder SH (1979) Immunohistochemical mapping of enkephalin containing cell bodies, fibers and nerve terminal in the brain stem of the rat. Brain Res 215: 235–255
Verhofstad AAJ, Steinbusch HWM, Joosten HWJ, Penke B, Barga J, Goldstein M (1982) Immunocytochemical localization of noradrenaline, adrenaline, and serotonin. In: Polack JM, Van Noorden S (eds) Immunocytochemistry: practical applications in pathology and biology. Wright, Bristol, pp 143–168
Wamsley JK, Young WS III, Kuhar MJ (1980) Immunohistochemical localization of enkephalin in rat forebrain. Brain Res 190: 153–174
Wessendorf M, Elde R (1985) Characterization of an immunohistochemical technique for the demonstration of co-existing neurotransmitters within nerve fibers and terminals. J Histochem Cytochem 33: 984–994
Willis WD, Haber LH, Martin RF (1977) Inhibition of spinothalamic tract cells and interneurons by brainstem stimulation in the monkey. J Neurophysiol 40: 968–981
Zamboni L, de Martino C (1967) Buffered picric acid-formaldehyde: a new rapid fixative for electron microscopy. J Cell Biol 148A: 35
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Millhorn, D.E., Hökfelt, T., Verhofstad, A.A.J. et al. Individual cells in the raphe nuclei of the medulla oblongata in rat that contain immunoreactivities for both serotonin and enkephalin project to the spinal cord. Exp Brain Res 75, 536–542 (1989). https://doi.org/10.1007/BF00249904
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DOI: https://doi.org/10.1007/BF00249904