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GABA neurons in the rat suprachiasmatic nucleus: Involvement in chemospecific synaptic circuitry and evidence for GAD-peptide colocalization

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Journal of Neurocytology

Summary

Dual labelling methods were employed for the electron microscopic detection of glutamate decarboxylase (GAD) immunoreactivity, together with vasoactive intestinal peptide (VIP) or neuropeptide Y (NPY) immunoreactivity in the suprachiasmatic nucleus (SCN) of colchicine pretreated and untreated rats. These methods involved the combined use of diaminobenzidine and benzidine dihydrochloride as distinct chromogens to visualize peroxidase-anti-peroxidase (PAP) immunostaining, and a combination of the PAP procedure with a radioimmunocytochemical method employing125I-labelled secondary antisera. We were thereby able to demonstrate that γ-aminobutyric acid (GABA) terminals provide an important afferent synaptic input to VIP neurons. Some of these VIP-immunoreactive neurons also exhibited GAD immunoreactivity. Examples of direct appositions between GABA and NPY terminals, and of a convergence of the two types of terminals on to the same postsynaptic targets, were frequently encountered. NPY/GAD colocalization within a few axonal varicosities was also demonstrated. These data provide additional information concerning chemospecific neuronal interactions that could be of functional importance in the regulation of circadian rhythmicity at the level of the SCN.

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References

  • Albers, H. E., Minamitani, N., Stopa, E. &Ferris, C. F. (1987) Light selectively alters vasoactive intestinal peptide and peptide histidine isoleucine immunoreactivity within the rat suprachiasmatic nucleus.Brain Research 437, 189–92.

    PubMed  Google Scholar 

  • Andre, D., Vuillon-Cacciuttolo, G. &Bosler, O. (1987) GABA nerve endings in the rat red nucleus. Combined detection with serotonin terminals using dual immunocytochemistry.Neuroscience 23, 1095–102.

    PubMed  Google Scholar 

  • Aoki, C., &Pickel, V. M. (1989) Neuropeptide Y in the cerebral cortex and the caudate-putamen nuclei. Ultrastructural basis for interactions with GABAergic and non-GABAergic neurons.Journal of Neuroscience 9, 4333–54.

    Google Scholar 

  • Bosler, O. (1989) Ultrastructural relationships of serotonin and GABA terminals in the rat suprachiasmatic nucleus. Evidence for a close interconnection between the two afferent systems.Journal of Neurocytology 18, 105–13.

    PubMed  Google Scholar 

  • Bosler, O. &Beaudet, A. (1985) VIP neurons as prime synaptic targets for serotonin afferents in rat suprachiasmatic nucleus: a combined radioautographic and immunocytochemical study.Journal of Neurocytology 14, 749–63.

    PubMed  Google Scholar 

  • Card, J. P., Brecha, N., Karten, H. J. &Moore, R. Y. (1981) Immunocytochemical localization of vasoactive intestinal polypeptide-containing cells and processes in the suprachiasmatic nucleus of the rat: light and electron microscopic analysis.Journal of Neuroscience 1, 1289–303.

    PubMed  Google Scholar 

  • Card, J. P., Fitzpatrick-McElligott, S., Gozes, I. &Baldino, F. (1988) Localization of vasopressin-, vasoactive intestinal polypeptide-, peptide histidine isoleucine and somatostatin-mRNA in rat suprachiasmatic nucleus.Cell and Tissue Research 252, 307–15.

    PubMed  Google Scholar 

  • Card, J. P. &Moore, R. Y. (1982) Ventral lateral geniculate nucleus afferents to the rat suprachiasmatic nucleus exhibit avian pancreatic polypeptide-like immunoreactivity.Journal of Comparative Neurology 206, 390–6.

    PubMed  Google Scholar 

  • Card, J. P. &Moore, R. Y. (1988) Neuropeptide Y localization in the rat suprachiasmatic nucleus and periventricular hypothalamus.Neuroscience Letters 88, 241–6.

    PubMed  Google Scholar 

  • Card, J. P. &Moore, R. Y. (1989) Organization of lateral geniculate-hypothalamic connections in the rat.Journal of Comparative Neurology 284, 134-A7.

    Google Scholar 

  • De Quidt, M. E. &Emson, P. C. (1986) Distribution of neuropeptide Y-like immunoreactivity in the rat central nervous system. II. Immunohistochemical analysis.Neuroscience 18, 545–618.

    PubMed  Google Scholar 

  • Francois-Bellan, A. M., Hery, M., Barrit, M. C., Faudon, M. &Hery, F. (1987) The stimulation of GABAB receptors increases serotonin release in the rat suprachiasmatic area.Neurochemistry International 11, 55–62.

    Google Scholar 

  • Francois-Bellan, A. M., Hery, M., Faudon, M. &Hery, F. (1988) Evidence for GABA control of serotonin metabolism in the rat suprachiasmatic area.Neurochemistry International 13, 455–62.

    Google Scholar 

  • Guy, J., Bosler, O., Dusticier, G. Pelletier, G. &Calas, A. (1987) Morphological correlates of serotonin-neuropeptide Y interactions in the rat suprachiasmatic nucleus: combined radioautographic and immunocyto-chemical data.Cell and Tissue Research 250, 657–62.

    PubMed  Google Scholar 

  • Harrington, M. E., Nance, D. M. &Rusak, B. (1987) Double-labelling of neuropeptide Y-immunoreactive neurons which project from the geniculate to the suprachiasmatic nuclei.Brain Research 410, 275–82.

    PubMed  Google Scholar 

  • Hendry, S. H. C., Jones, E. G., DeFelipe, J., Schemechel, D., Brandon, C. &Emson, P. C. (1984) Neuropeptide-containing neurons of the cerebral cortex are also GABA-ergic.Proceedings of the National Academy of Sciences (USA) 81, 6526–30.

    Google Scholar 

  • Hisano, S., Chikamori-Aoyama, M., Katoh, S., Kagotani, Y., Daikoku, S. &Chihara, K. (1988) Suprachiasmatic nucleus neurons immunoreactive for vasoactive intestinal polypeptide have synaptic contacts with axons immunoreactive for neuropeptide Y: an immunoelectron microscopic study in the rat.Neuroscience Letters 88, 145–50.

    PubMed  Google Scholar 

  • Ibata, Y., Takahashi, Y., Okamura, H., Kawakami, F., Terubayashi, H., Kubo, T. &Yanaihara, N. (1989) Vasoactive intestinal peptide (VIP)-like immunoreactive neurons located in the rat suprachiasmatic nucleus receive a direct retinal projection.Neuroscience Letters 97, 1–5.

    PubMed  Google Scholar 

  • Jegou, S., Javoy-Agid, F., Delbende, C., Tranchand-Bunel, D., Coy, D. H., Agid, Y. &Vaudry, H. (1988) Regional distribution of vasoactive intestinal peptide in brains from normal and parkinsonian subjects.Peptides 9, 787–93.

    PubMed  Google Scholar 

  • Kachidian, P., Denoroy, L. &Bosler, O. (1989) Adrenergic neurons in the nucleus tractus solitarius receive GABAergic synapses. Demonstration by dual immuno-cytochemistry in the rat.Neuroscience Letters 100, 65–70.

    PubMed  Google Scholar 

  • Kiss, J., Leranth, C. &Halasz, B. (1984) Serotoninergic endings on VIP-neurons in the suprachiasmatic nucleus and on ACTH-neurons in the arcuate nucleus of the rat hypothalamus. A combination of high resolution auto-radiography and electron microscopic immunocyto-chemistry.Neuroscience Letters 44, 119–24.

    PubMed  Google Scholar 

  • Levey, A. I., Bolam, J. P., Rye, D. B., Hallanger, A. E., Demuth, R. M., Mesulam, M. M. &Wainer, B. H. (1986) A light and electron microscopic procedure for sequential double antigen localization using diaminobenzidine and benzidine dihydrochloride.Journal of Histochemistry and Cytochemistry 34, 1449–57.

    PubMed  Google Scholar 

  • McDonald, A. J. &Pearson, J. C. (1989) Coexistence of GABA and peptide immunoreactivity in non-pyramidal neurons of the basolateral amygdala.Neuroscience Letters 100, 53–8.

    PubMed  Google Scholar 

  • McLean, S., Skirboll, L. R. &Pert, C. B. (1983) Opiatergic projection from the bed nucleus of the habenula: demonstration by a novel radioimmunohistochemical method.Brain Research 278, 255–7.

    PubMed  Google Scholar 

  • Meijer, J. H. &Rietveld, W. J. (1989) Neurophysiology of the suprachiasmatic circadian pacemaker in rodents.Physiological Reviews 69, 671–707.

    PubMed  Google Scholar 

  • Milner, T. A., Pickel, V. M., Chan, J., Massari, V. J., Oertel, W. H., Park, D. H., Joh, T. H. &Reis, D. J. (1987) Phenylethanolamine N-methyltransferase-containing neurons in the rostral ventrolateral medulla. II. Synaptic relationships with GABAergic terminals.Brain Research 411, 46–57.

    PubMed  Google Scholar 

  • Mugnaini, E. &Oertel, W. H. (1985) An atlas of the distribution of GABAergic neurons and terminals in the rat CNS as revealed by GAD immunohistochemistry. InHandbook of Chemical Neuroanatomy Vol. 4:GABA and Neuropeptides in the CNS. Part I (edited byBjorklund, A. &Hokfelt, T.) pp. 436–595. Amsterdam: Elsevier.

    Google Scholar 

  • Oertel, W. H., Schmechel, D. E., Daly, J. W., Tappaz, M. L. &Kopin, I. J. (1980) Localization of glutamate decar-boxylase on line-immunoelectrophoresis and two- dimensional electrophoresis by use of the radioactive suicide substrate 2 [3H]gamma-acetylenic GABA.Life Science 27, 2133–41.

    Google Scholar 

  • Oertel, W. H., Schmechel, D. E., Tappaz, M. L. &Kopin, I. J. (1981) Production of a specific antiserum to rat brain glutamic acid decarboxylase (GAD) by injection of an antigen-antibody complex.Neuroscience 6, 2689–700.

    PubMed  Google Scholar 

  • Ohara, P. T., Lieberman, A. R., Hunt, S. P. &Wu, J. Y. (1983) Neural elements containing glutamic acid decarboxylase (GAD) in the dorsal lateral geniculate nucleus of the rat. Immunohistochemical studies by light and electron microscopy.Neuroscience 8, 189–211.

    PubMed  Google Scholar 

  • Okamura, H., Berod, A., Julien, J. F., Geffard, M., Kitahama, K., Mallet, J. &Bobillier, P. (1989). Demonstration of GABAergic cell bodies in the supra-chiasmatic nucleus: in situ hybridization of glutamic acid decarboxylase (GAD) mRNA and immunocytochemistry of GAD and GABA.Neuroscience Letters 102, 131–6.

    PubMed  Google Scholar 

  • Pelletier, G., Desy, L., Kerkerian, L. &Cote, J. (1984) Immunocytochemical localization of neuropeptide Y (NPY) in the human hypothalamus.Cell and Tissue Research 238, 203–5.

    PubMed  Google Scholar 

  • Ralph, M. R., &Menaker, M. (1985) Bicuculline blocks circadian phase delays but not advances.Brain Research 325, 363–5.

    Google Scholar 

  • Ralph, M. R. &Menaker, M. (1986). Effects of diazepam on circadian phase advances and delays.Brain Research 372, 405–8.

    PubMed  Google Scholar 

  • Ralph, M. R. &Menaker, M. (1989) GABA regulation of circadian responses to light. I. Involvement of GABAA- benzodiazepine and GABAB receptors.Journal of Neuroscience 9, 2858–65.

    PubMed  Google Scholar 

  • Sakaue, M. Saito, N., Taniguchi, H., Baba, S. &Tanaka, C. (1988). Immunohistochemical localization of γ-aminobutyric acid in the rat pituitary gland and related hypothalamic regions.Brain Research 446, 343–53.

    PubMed  Google Scholar 

  • Smith, Y. Parent, A., Kerkerian, L. &Pelletier, G. (1985) Distribution of neuropeptide Y Immunoreactivity in the basal forebrain and upper brainstem of the squirrel monkey (Saimiri sciureus).Journal of Comparative Neurology 236, 71–89.

    PubMed  Google Scholar 

  • Tappaz, M. L., Bosler, O., Paut, L. &Berod, A. (1985) Glutamate decarboxylase-immunoreactive boutons in synaptic contacts with hypothalamic dopaminergic cells: a light and electron microscopy study combining immunocytochemistry and radioautography.Neuroscience 16, 111–22.

    PubMed  Google Scholar 

  • Tappaz, M. L., Wassef, M., Oertel, W. H., Paut, L. &Pujol, J. F. (1983) Light- and electron-microscopic immunocytochemistry of glutamic acid decarboxylase (GAD) in the basal hypothalamus: morphological evidence for neuroendocrine γ-ammobutyrate (GABA).Neuroscience 9, 271–87.

    PubMed  Google Scholar 

  • Turek, F. W. &Losee-Olson, S. (1986) A benzodiazepine used in the treatment of insomnia phase-shifts the mammalian circadian clock.Nature 321, 167–8.

    PubMed  Google Scholar 

  • Ueda, S., Kawata, M. &Sano, Y. (1986) Identification of neuropeptide Y immunoreactivity in the suprachiasmatic nucleus and the lateral geniculate nucleus of some mammals.Neuroscience Letters 68, 7–10.

    PubMed  Google Scholar 

  • Van Den Pol, A. N. (1980). The hypothalamic suprachiasmatic nucleus of the rat: intrinsic anatomy.Journal of Comparative Neurology 191, 661–702.

    PubMed  Google Scholar 

  • Van Den Pol, A. N. (1986) Gamma-aminobutyrate, gastrin releasing peptide, serotonin, somatostatin, and vaso- pressin: ultrastructural immunocytochemical localization in presynaptic axons in the suprachiasmatic nucleus.Neuroscience 17, 643–59.

    PubMed  Google Scholar 

  • Van Den Pol, A. N. &Gorcs, T. (1986) Synaptic relationships between neurons containing vasopressin, gastrin- releasing peptide, vasoactive intestinal polypeptide, and glutamate decarboxylase immunoreactivity in the suprachiasmatic nucleus: dual ultrastructural immunocytochemistry with gold-substituted silver peroxidase.Journal of Comparative Neurology 252, 507–21.

    PubMed  Google Scholar 

  • Van Den Pol, A. N. &Tsujimoto, K. L. (1985) Neurotrans-mitters of the hypothalamic suprachiasmatic nucleus: immunocytochemical analysis of 25 neuronal antigens.Neuroscience 15, 1049–86.

    PubMed  Google Scholar 

  • Van Reeth, O., Losee-Olson, S. &Turek, F. W. (1987). Phase shifts in the circadian activity rhythm, induced by triazolam are not mediated by the eyes or the pineal gland in the hamster.Neuroscience Letters 80, 185–90.

    PubMed  Google Scholar 

  • Vincent, S. R., Hökfelt, T. &Wu, J. Y. (1982) GABA neuron systems in hypothalamus and the pituitary gland. Immunohistochemical demonstration using antibodies against glutamate decarboxylase.Neuroendocrinology 34, 117–25.

    PubMed  Google Scholar 

  • Vuillet, J., Kerkerian-Le Goff, L., Kachidian, P., Dusticier, G., Bosler, O. &Nieoullon, A. (1990). Striatal NPY-containing neurons receive GABAergic afferents and may also contain GABA: an electron microscopic study in the rat.European Journal of Neuroscience 2.

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Francois-Bellan, A.M., Kachidian, P., Dusticier, G. et al. GABA neurons in the rat suprachiasmatic nucleus: Involvement in chemospecific synaptic circuitry and evidence for GAD-peptide colocalization. J Neurocytol 19, 937–947 (1990). https://doi.org/10.1007/BF01186821

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  • DOI: https://doi.org/10.1007/BF01186821

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