Localization of Dopamine D2 Receptor in Rat Spinal Cord Identified with Immunocytochemistry and In Situ Hybridization
Corresponding Author
Henk van Dijken
Department of Anatomy, Erasmus University Medical School, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Correspondence to: Henk van Dijken, as aboveSearch for more papers by this authorJan Dijk
Department of Medical Biochemistry, Sylvius Laboratory, University of Leiden, The Netherlands
Search for more papers by this authorPieter Voorn
Department of Anatomy and Embryology, Free University, Amsterdam, The Netherlands
Search for more papers by this authorJan C. Holstege
Department of Anatomy, Erasmus University Medical School, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Search for more papers by this authorCorresponding Author
Henk van Dijken
Department of Anatomy, Erasmus University Medical School, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Correspondence to: Henk van Dijken, as aboveSearch for more papers by this authorJan Dijk
Department of Medical Biochemistry, Sylvius Laboratory, University of Leiden, The Netherlands
Search for more papers by this authorPieter Voorn
Department of Anatomy and Embryology, Free University, Amsterdam, The Netherlands
Search for more papers by this authorJan C. Holstege
Department of Anatomy, Erasmus University Medical School, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Search for more papers by this authorAbstract
In the present study the distribution of dopamine D2 receptors in rat spinal cord was determined by means of immunocytochemistry using an anti-peptide antibody, directed against the putative third intracellular loop of the D2 receptor and in situ hybridization (ISH) using a [35S]UTP labelled anti-sense riboprobe. With the immunocytochemical technique, labelling was confined to neuronal cell bodies and their proximal dendrites. Strongest labelling was present in the parasympathetic area of the sacral cord and in two sexually dimorphic motor nuclei of the lumbosacral cord, the spinal nucleus of the bulbocavernosus and the dorsolateral nucleus. Moderately labelled cells were present in the intermediolateral cell column, the area around the central canal and lamina I of the dorsal horn. Weak labelling was present in the lateral spinal nucleus and laminae VII and VIII of the ventral horn. Except for the two sexually dimorphic motornuclei of the lumbosacral cord labelled motoneurons were not encountered. With the ISH technique radioactive labelling was present in many neurons, indicating that they contained D2 receptor mRNA. The distribution of these neurons was very similar to the distribution obtained with immunocytochemistry, but with ISH additional labelled cells were detected in laminae III and IV of the dorsal horn, which were never labelled with immunocytochemistry. The present study shows that the D2 receptor is expressed in specific areas of the rat spinal cord. This distribution provides anatomical support for the involvement of D2 receptors in modulating nociceptive transmission and autonomic control. Our data further indicate that D2 receptors are not directly involved in modulating motor functions with the exception, possibly, of some sexual motor functions.
Abbreviations:
-
- DA
-
- dopamine(ergic)
-
- DLN
-
- dorsolateral nucleus
-
- IML
-
- intermediolateral cell column
-
- SNB
-
- spinal nucleus of the bulbocavernosus
References
- Ariano, M. A., Fisher, R. S., Smyk-Randall, E., Sibley, D. R. and Levine, M. S. (1993) D2 dopamine receptor distribution in the rodent CNS using anti-peptide antisera. Brain Res., 609, 71–80.
- Ariano, M. A. and Sibley, D. R. (1994) Dopamine receptor distribution in the rat CNS: elucidation using anti-peptide antisera directed against D1A and D3 subtypes. Brain Res., 649, 95–110.
- Barasi, S., Ben-Sreti, M. M., Clatworthy, A. L., Duggal, K. N., Gonzalez, J. P., Robertson, J., Rooney, K. F. and Sewell, R. D. E. (1987) Dopamine receptor-mediated spinal antinociception in the normal and haloperidol pretreated rat: effects of sulpiride and SCH 23390. Br. J. Pharmacol., 90, 15–22.
- Barasi, S. and Roberts, M. H. T. (1977) Responses of motoneurones to electrophoretically applied dopamine. Br. J. Pharmacol., 60, 29–34.
- Bhargava, H. N. and Gulati, A. (1990) Modification of brain and spinal cord dopamine D1 receptors labeled with [3H]SCH 23390 after morphine withdrawal from tolerant and physically dependent rats. J. Pharmacol. Exp. Ther., 252, 901–907.
- Björklund, A. and Skagerberg, G. (1979) Evidence for a major spinal cord projection from the diencephalic A11 dopamine cell group in the rat using transmitter-specific fluorescent retrograde tracing. Brain Res., 177, 170–175.
- Boundy, V. A., Luedtke, R. R., Artymyshyn, R. P., Filtz, T. M. and Molinoff, P. B. (1993a) Development of polyclonal anti-D2 dopamine receptor antibodies using sequence-specific peptides. Mol. Pharmacol., 43, 666–676.
- Boundy, V. A., Luedtke, R. R. and Molinoff, P. B. (1993b) Development of polyclonal anti-D2 dopamine receptor antibodies to fusion proteins—inhibition of D2 receptor-G protein interaction. J. Neurochem., 60, 2181–2191.
- Bouthenet, M.-L., Martres, M.-P. Sales, N. and Schwartz, J.-C. (1987) A detailed mapping of dopamine D-2 receptors in rat central nervous system by autoradiography with [125I]iodosulpiride. Neuroscience, 20, 117–155.
- Bouthenet, M.-L., Souil, E., Martres, M.-P., Sokoloff, P., Giros, B. and Schwartz, J.-C. (1991) Localization of dopamine D3 receptor mRNA in the rat brain using in situ hybridization histochemistry: comparison with dopamine D2 receptor mRNA. Brain Res., 564, 203–219.
- Brock, J. W., Farooqui, S., Ross, K. and Prasad, C. (1992) Localization of dopamine D2 receptor protein in rat brain using polyclonal antibody. Brain Res., 578, 244–250.
- Brouwer, N., van Dijken, H., Ruiters, M. H. J., van Willigen, J.-D. and Ter Horst, G. J. (1992) Localization of dopamine D2 receptor mRNA with non-radioactive in situ hybridization histochemistry. Neurosci. Lett., 142, 223–227.
- Bunzow, J. R., Van Tol, H. H. M., Grandy, D. K., Albert, P., Salon, J., Christie, M., Machida, C. A., Neve, K. A. and Civelli, O. (1988) Cloning and expression of a rat D2 dopamine receptor cDNA. Nature, 336, 783–787.
- Carp, J. S. and Anderson, R. J. (1982) Dopamine receptor-mediated depression of spinal monosynaptic transmission. Brain Res., 242, 247–254.
- Chazot, P. L., Doherty, A. J. and Strange, P. G. (1993) Antisera specific for D2 dopamine receptors. Biochem. J., 289, 789–794.
- Civelli, O., Bunzow, J. R., Grandy, D. K., Zhou, Q.-Y. and Van Tol, H. H. M. (1991) Molecular biology of the dopamine receptors. Eur. J. Pharmacol.—Mol. Pharmacol. Section, 207, 277–286.
- Clatworthy, A. and Barasi, S. (1987) Intrathecally administered apomorphine or LY171555 reduces nociceptive responses recorded from ventrobasal thalamic neurones in urethane anaesthetised rats. Neurosci. Lett., 75, 308–312.
- Commissiong, J. W. and Neff, N. H. (1979) Current status of dopamine in the mammalian spinal cord. Biochem. Pharmacol., 28, 1569–1573.
- Coote, J. H., Macleod, V. H., Fleetwood-Walker, S. and Gilbey, M. P. (1981) The response of individual sympathetic preganglionic neurones to microelectrophoretically applied endogenous monoamines. Brain Res., 215, 135–145.
- Demenge, P., Feuerstein, C., Mouchet, P. and Guerin, B. (1980) Stereospecific binding of 3H-haloperidol in rat dorsal spinal cord. Eur. J. Pharmacol., 66, 117–120.
- Dubois, A., Savasta, M., Curet, O. and Scatton, B. (1986) Autoradiographic distribution of the D1 agonist [3H]SKF 38393, in the rat brain and spinal cord. Comparison with the distribution of D2 dopamine receptors. Neuroscience, 19, 125–137.
- Fisher, R. S., Levine, M. S., Sibley, D. R. and Ariano, M. A. (1994) D2 dopamine receptor protein location: golgi impregnation-gold toned and ultrastructural analysis of the rat neostriatum. J. Neurosci. Res., 38, 551–564.
- Fleetwood-Walker, S. M., Hope, P. J. and Mitchell, R. (1988) Antinociceptive actions of descending dopaminergic tracts on cat and rat dorsal horn somatosensory neurones. J. Physiol. (Lond.), 399, 335–348.
- Gingrich, J. A. and Caron, M. G. (1993) Recent advances in the molecular biology of dopamine receptors. Annu. Rev. Neurosci., 16, 299–321.
- Hökfelt, T., Phillipson, O. and Goldstein, M. (1979) Evidence for a dopaminergic pathway in the rat descending from the A11 cell group to the spinal cord. Acta Physiol. Scand., 107, 393–395.
- Holstege, J. C., Goedknegt, H., Bongers, C. M. H., van Veen, T. and Buijs, R. M. (1990) Light microscopical identification of dopamine immuno-reactivity in the rat, cat and monkey spinal cord. Eur. J. Neurosci., Suppl. 3, 228.
- Jongen-Rêlo, A. L., Docter, G. J., Jonker, A. J., Vreugdenhil, E., Groenewegen, H. J. and Voorn, P. (1994) Differential effects of dopamine depletion on the binding- and mRNA-levels of dopamine receptors in the shell and core of the rat nucleus accumbens. Mol. Brain Res., 25, 333–343.
- Kamijo, N., Nagao, T. and Ono, H. (1993) Depression of the monosynaptic reflex by apomorphine or bromocryptine is not mediated by D1/D2 receptors. Neuropharmacology, 32, 777–783.
- Lahlou, S. and Demenge, P. (1993) Cardiovascular responses to intrathecal dopamine receptor agonists after spinal transection in conscious rats. Cardiovasc. Res., 27, 222–230.
- Levey, A. I., Hersch, S. M., Rye, D. B., Sunahara, R. K., Niinik, H. B., Kitt, C. A., Price, D. L., Maggio, R., Brann, M. R. and Ciliax, B. J. (1993) Localization of D1 and D2 dopamine receptors in brain with subtype-specific antibodies. Proc. Natl Acad. Sci. USA, 90, 8861–8865.
- Liu, Q.-S., Qiao, J.-T. and Dafny, N. (1992) D2 dopamine receptor involvement in spinal dopamine-produced antinociception. Life Sci., 51, 1485–1492.
- Magnusson, T. (1973) Effect of chronic transection on dopamine, noradrenaline and 5-hydroxytryptamine in the rat spinal cord. Naunyn Schmiedebergs Arch. Pharmacol., 278, 13–22.
- Mouchet, P., Manier, M. and Feuerstein, C. (1992) Immunohistochemical study of the catecholaminergic innervation of the spinal cord of the rat using specific antibodies against dopamine and noradrenaline. J. Chem. Neuroanat., 5, 427–440.
- Ono, H. and Fukuda, H. (1984) Effect of methamphetamine on rat spinal cord. Dopamine receptor-mediated depression of monosynaptic reflex. Neuropharmacology, 23, 637–642.
- Pehek, E. A., Thompson, J. T. and Hull, E. M. (1989) The effects of intrathecal administration of the dopamine angonist apomorphine on penile reflexes and copulation in the male rat. Psychopharmacology (Berl.), 99, 304–308.
- Pellissier, G. and Demenge, P. (1991) Hypotensive and bradycardic effects elicited by spinal dopamine receptor stimulation: effects of D1 and D2 receptor agonists and antagonists. J. Cardiovasc. Pharmacol., 18, 548–555.
- Petitjean, P., Mouchet, P., Pellissier, G., Manier, M., Feuerstein, C. and Demenge, P. (1984) Cardiovascular effects in the rat of intrathecal injections of apomorphine at the thoracic spinal cord level. Eur. J. Pharmacol., 105, 355–359.
- Plug, M. J., Dijk, J., Maassen, J. A. and Möller, W. (1992) An anti-peptide antibody that recognizes the dopamine D2 receptor from bovine striatum. Eur. J. Biochem., 206, 123–130.
- Ridet, J.-L., Sandillon, F., Rajaofetra, N., Geffard, M. and Privat, A. (1992) Spinal dopaminergic system of the rat: light and electron microscopic study using an antiserum against dopamine, with particular emphasis on synaptic incidence. Brain Res., 598, 233–241.
- Scatton, B., Dubois, A. and Cudennec, A. (1984) Autoradiographic localization of dopamine receptors in the spinal cord of the rat using [3H]-N-propylnorapomorphine. J. Neural. Transm., 59, 251–256.
- Sesack, S. R., Aoki, C. and Pickel, V. M. (1994) Ultrastructural localization of D2 receptor-like immunoreactivity in midbrain dopamine neurons and their striatal targets. J. Neurosci., 14, 88–106.
- Shirouzu, M., Anraku, T., Iwashita, Y. and Yoshida, M. (1990) A new dopaminergic terminal plexus in the ventral horn of the rat spinal cord. Immunohistochemical studies at the light and electron microscopic levels. Experientia, 46, 201–204.
- Skagerberg, G., Björklund, A., Lindvall, O. and Schmidt, R. H. (1982) Origin and termination of the diencephalo-spinal dopamine system in the rat. Brain Res. Bull., 9, 237–244.
- Skagerberg, G., Meister, B., Hökfelt, T., Lindvall, O., Goldstein, M., Joh, T. and Cuello, A. C. (1988) Studies on dopamine-, tyrosine hydroxylase- and aromatic L-amino acid decarboxylase-containing cells in the rat diencephalon: comparison between formaldehyde-induced histofluorescence and immunofluorescence. Neuroscience, 24, 605–620.
- Smiley, J. F., Levey, A. I., Ciliax, B. J. and Goldman-Rakic, P. S. (1994) D1 dopamine receptor immunoreactivity in human and monkey cerebral cortex: Predominant and extrasynaptic localization in dendritic spines. Proc. Natl Acad. Sci., 91, 5720–5724.
- Smith, D. O., Lowe, D., Temkin, R., Jensen, P. and Hatt, H. (1995) Dopamine enhances glutamate-activated currents in spinal motoneurons. J. Neurosci., 15, 3905–3912.
- Stefanick, M. L., Smith, E. R., Clark, J. T. and Davidson, J. M. (1982) Effects of a potent dopamine receptor agonist, RDS-127, on penile reflexes and seminal emission in intact and spinally transected rats. Physiol. Behav., 29, 973–978.
- Swanson, L. W., Sawchenko, P. E., Berod, A., Hartman, B. K., Helle, K. B. and Vanorden, D. E. (1981) An immunohistochemical study of the organization of catecholaminergic cells and terminal fields in the paraventricular and supraoptic nuclei of the hypothalamus. J. Comp. Neurol., 196, 271–285.
- Weiner, D. M., Levey, A. I., Sunahara, R. K., Niznik, H. B., O'Dowd, B. F., Seeman, P. and Brann, M. R. (1991) D1 and D2 dopamine receptor mRNA in rat brain. Proc. Natl. Acad. Sci., 88, 1859–1863.
- Yokoyama, C., Okamura, H., Nakajima, T., Taguchi, J. I. and Ibata, Y. (1994) Autoradiographic distribution of [3H]YM-09151–2, a high-affinity and selective antagonist ligand for the dopamine D2 receptor group, in the rat brain and spinal cord. J. Comp. Neurol., 344, 121–136.