Summary
The extensive distribution of exohypothalamic vasopressin or oxytocin containing nerve fibres is thought to be the anatomical basis for the involvement of these neuropeptides in central processes. Following light microscopic observations suggesting that these fibres terminate on other neurons, the present study was undertaken to demonstrate the existence of such endings in the limbic system, which is one of the main target areas for these peptides. For immunoelectron microscopy glutaraldehyde-paraformaldehyde perfused brains of male Wistar rats and Brattleboro rats, homozygous for diabetes insipidus, with and without postfixation in OsO4, were used. Post-embedding staining revealed false positive reaction product on all dense core vesicles, e.g., in the lateral septum. With pre-embedding staining, however, intense and specific reactions were observed for both vasopressin and oxytocin at their sites of production, as well as the neurohypophysis and in the extrahypothalamic limbic brain regions.
In the lateral septum and habenular nucleus only vasopressin-containing synapses could be demonstrated, while in the medial nucleus of the amygdala synapses containing either vasopressin or oxytocin were observed. These peptide containing synapses do not seem to differ in any fundamental way from the classical transmitter-containing synapses in the brain.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alonso, G., Assenmacher, I.: The smooth endoplasmic reticulum in neurosecretory axons of the rat neurohypophysis. Biol. Cell 32, 203–206 (1978)
Andersson, B., Leksell, L.G., Lishajko, F.: Perturbation in fluid balance induced by medially placed forebrain lesions. Brain Res. 99, 261–275 (1975)
Barchas, J.D., Akil, H., Elliott, G.R., Holman, R.B., Watson, S.J.: Behavioral neurochemistry: Neuroregulators and behavioral states. Science 200, 964–973 (1978)
Barker, J.L.: Physiological roles of peptides in the nervous system. In: Peptides in neurobiology (H. Gainer, ed.) pp. 295–343, New York: Plenum Press, 1977
Bohus, B., Urban, I., Wimersma Greidanus, T.J.B. van, Wied, D. de: Opposite effects of oxytocin and vasopressin on avoidance behavior and hippocampal theta rhythms in the rat. Neuropharmacology 17, 239–248 (1978)
Buijs, R.M.: Intra- and extrahypothalamic vasopressin and oxytocin pathways in the rat; pathways to the limbic system, medulla oblongata and spinal cord. Cell Tissue Res. 192, 423–435 (1978a)
Buijs, R.M.: The specific localization of vasopressin and oxytocin pathways in the rat central nervous system. In: Hormones and Brain Developments (G. Dörner and M. Kawakami, eds.) pp. 223–228, Amsterdam: Elsevier North Holland Biomedical Press, 1978b
Buijs, R.M., Swaab, D.F., Dogterom, J., Leeuwen, F.W. van: Intra- and extrahypothalamic vasopressin and oxytocin pathways in the rat. Cell Tissue Res. 186, 423–433 (1978)
Dogterom, J., Snijdewint, F.G.M., Buijs, R.M.: The distribution of vasopressin and oxytocin in the rat brain. Neurosci. Lett. 9, 341–346 (1978)
Droz, B., Koenig, H.L., Giaumberardino, L.J.: Axonal migration of protein and glycoprotein to nerve endings: I. Radioautographic analysis of the renewal of protein in nerve endings of chicken ciliary ganglions after intracerebral injection of 3H-Lysine. Brain Res. 60, 93–127 (1973)
Droz, B., Rambourg, A., Koenig, H.L.: The smooth endoplasmatic reticulum: structure and role in the renewal of axonal membrane and synaptic vesicles by fast axonal transport. Brain res. 93, 1–13 (1975)
Dube, D., Leclerc, R., Pelletier, G.: Electronmicroscopic immunohistochemical localization of vasopressin and neurophysin in the median eminence of normal and adrenalectomized rats. Am. J. Anat. 146, 103–108 (1976)
Flament-Durand, J.: Ultrastructural aspects of the paraventricular nuclei in the rat. Z. Zellforsch. 116, 61–69 (1971)
Heller, H., Hasan, S.A., Saifi, A.Q.: Antidiuretic activity in the cerebrospinal fluid. J. Endocrinol. 41, 273–280 (1968)
Johnson, A.K., Buggy, J.: Periventricular preoptic hypothalamus is vital for thirst and normal water economy. Am. J. Physiol. 234, R122-R129 (1978)
Kovacs, G.L., Bohus, B., Versteeg, D.H.G., Kloet, E.R. de, Wied, D. de: Effect of oxytocin and vasopressin on memory consolidation: sites of action and catecholaminergic correlates after local microinjection into limbic-midbrain structures. Brain Res., in press.
Leeuwen, F.W. van, Swaab, D.F.: Specific immunoelectronmicroscopic localization of vasopressin and oxytocin in the neurohypophysis of the rat. Cell Tissue Res. 177, 493–501 (1977)
Livingston, A.: Subcellular aspects of storage and release of neurohypophysial hormones. J. Endocrinol 49, 357–372 (1971)
Negoro, H., Visessuwan, S., Holland, R.C.: Inhibition and excitation of units in paraventricular nucleus after stimulation of septum amygdala and neurohypophysis. Brain Res. 57, 479–483 (1973)
Ono, T., Nishino, H., Sasaka, K., Muramoto, K., Yano, I., Simpson, A.: Paraventricular nucleus connections to spinal cord and pituitary. Neurosci. Lett. 10, 141–146 (1978)
Pittman, Q.J., Blume, H.W., Renaud, L.P.: Afferent and efferent connections of putative peptidergic neurons of the paraventricular nucleus (PVN). Soc. Neurosci. 4, 352 (1978)
Schober, F.: Darstellung der neurosekretorischen hypothalamo-rhombenzephalen Verbindung bei der Ratte durch retrograden axonalen Transport von Meerettich-Peroxidase. Acta Biol. Med. Germ. 37, 165–167 (1978)
Sternberger, L.A.: Immunocytochemistry, New York: Prentice Hall, Englewood Cliffs, 1974
Swaab, D.F., Pool, C.W.: Specificity of oxytocin and vasopressin immunofluorescence. J. Endocrinol 66, 263–272 (1975)
Swaab, D.F., Pool, C.W., Leeuwen, F.W. van: Can specificity ever be proved in immunocytochemical staining? J. Histochem. Cytochem. 25, 388–391 (1977)
Tanaka, M., Kloet, E.R. de, Wied, D. de, Versteeg, D.H.G.: Arginine8-vasopressin affects catecholamine metabolism in specific brain nuclei. Life Sci. 20, 1799–1808 (1977)
Termaat, A.: pers. communication
Vandesande, F., Dierickx, K., Mey, J. de: The origin of the vasopressinergic and oxytocinergic fibres of the external region of the median eminence of the rat hypophysis. Cell Tissue Res. 180, 443–452 (1977)
Versteeg, D.H.G., Kloet, E.R. de, Wimersma Greidanus, Tj. B. van, Wied, D. de: Vasopressin modulates the activity of catecholamine containing neurons in specific brain regions. Neurosci. Lett. 11, 69–73 (1979)
Vorherr, H., Bradbury, M.W.B., Hoghoughi, M., Kleeman, C.R.: Antidiuretic hormone in cerebrospinal fluid during endogenous and exogenous changes in its blood level. Endocrinology 83, 246–250 (1968)
Wied, D. de, Bohus, B.: Modulation of memory processes by neuropeptides of hypothalamic neurohypophyseal origin. In: Brain mechanisms in memory and learning: from the single neuron to man (M.A.B. Brazier, ed.) pp. 139–149, New York: Raven Press, 1979
Wimersma Greidanus, Tj. B. van, Croiset, G., Bakker, E., Bouman, H.: Amygdaloid lesions block the effect of neuropeptides (vasopressin ACTH4–10) on avoidance behavior. Physiol. Behav. 22, 291–295 (1979)
Zambrano, D., De Robertis, E.: The secretory cycle of supraoptic neurons in the rat. Z. Zellforsch. 73, 414–431 (1966)
Author information
Authors and Affiliations
Additional information
Supported by the Foundation for Medical Research FUNGO
The authors wish to thank Prof. Dr. A.H.M. Lohman for having made the vibratome available, and Miss C. de Raay for her expert technical assistance
Rights and permissions
About this article
Cite this article
Buijs, R.M., Swaab, D.F. Immuno-electron microscopical demonstration of vasopressin and oxytocin synapses in the limbic system of the rat. Cell Tissue Res. 204, 355–365 (1979). https://doi.org/10.1007/BF00233648
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00233648