Summary
Synthetic oxytocin and [8-arginine]-vasopressin conjugated to bovine thyroglobulin were used to induce specific antibodies in rabbits. The specificity of the anti-oxytocin serum, and the suitability of the anti-[8arginine]-vasopressin serum for the detection of [8-lysine]-vasopressin, was evaluated by immunofluorescent studies of the respective hormones bound to Sepharose 4B particles. Oxytocin and [8-lysine]-vasopressin were specifically localized in the paraventricular (PVN) and supraoptic (SON) nuclei of the pig hypothalamus using the immunoperoxidase staining technique.
After an examination of serial transverse and sagittal sections stained for either of the hormones we observed that:
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1.
In the rostral SON, oxytocin and vasopressin containing neurons were uniformly distributed;
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2.
In the caudal SON, most of the neurons contained oxytocin, but there were still a few ‘vasopressin’ neurons;
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3.
In the rostral PVN, the two hormones were evenly spread in neurons close to the third ventricle;
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4.
In the caudal PVN, the oxytocin and vasopressin containing neurons were differentially distributed, with ‘oxytocin’ neurons adjacent to the third ventricle, and ‘vasopressin’ neurons lateral to these and concentrated in the dorso-caudal PVN.
In the cells of the PVN, there was evidence that the distribution of oxytocin and vasopressin is similar to the distribution of porcine neurophysin-II and porcine neurophysin-I respectively. This similarity is consistent with the one hormone — one neurophysin concept in the pig.
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References
Burford, G.D., Jones, C.W., Pickering, B.T.: Tentative identification of a vasopressin-neurophysin and an oxytocin-neurophysin in the rat. Biochem J. 124, 809–813 (1971)
Choy, V.J., Watkins, W.B.: Immunocytochemical study of the hypothalamo-neurohypophysial system. II. Distribution of neurophysin, oxytocin and vasopressin in the normal and osmotically stimulated rat. Cell Tiss. Res. 180, 467–490 (1977)
Dean, C.R., Hope, C.B., Kažić, T.: Evidence for the storage of oxytocin with neurophysin-I and of vasopressin with neurophysin-II in separate neurosecretory granules. Brit. J. Pharmacol. 34, 192p (1968)
De Mey, J., Dierickx, K., Vandesande, F.: Immunohistochemical demonstration of neurophysin-I and neurophysin-II containing nerve fibres in the external region of the bovine median eminence. Cell Tiss. Res. 157, 517–519 (1975a)
De Mey, J., Dierickx, K., Vandesande, F.: Identification of neurophysin producing cells. III. Immunohistochemical demonstration of neurophysin-I producing neurons in the bovine infundibular nucleus. Cell Tiss. Res. 161, 219–224 (1975b)
De Mey, J., Vandesande, F., Dierickx, K.: Identification of the neurophysin-I and the neurophysin-II producing neurons in the bovine hypothalamus. Cell Tiss. Res. 153, 531–543 (1974)
Fawcett, C.P., Powell, A.E., Sachs, H.: Biosynthesis and release of neurophysin. Endocrinology 83, 1299–1310 (1968)
Johnston, C.I., Hutchinson, J.S., Morris, B.J., Dax, E.M.: Release and clearance of neurophysins and posterior pituitary hormone. Ann. N. Y. Acad. Sci. 248, 272–280 (1975)
Johnston, C.I., Pickup, J.C., Uttenthal, L.O., Hope, D.B.: The relationship of neurophysins to vasopressin and oxytocin in pigs. Proc. Aust. End. Soc. 15, 41 (1972)
Livett, B.G., Uttenthal, L.O., Hope, D.B.: Localization of neurophysin-II in the hypothalamo-neurohypophysial system of the pig by immunofluorescence histochemistry. Phil. Trans. B 261, 371–378 (1971)
Legros, J.J., Reynaert, R., Peeters, G.: Specific release of bovine neurophysin-II during arterial or venous haemorrhage in the cow. J. Endocr. 67, 297–302 (1975)
Pickup, J.C., Johnston, C.I., Nakamura, S., Uttenthal, L.O., Hope, D.B.: Subcellular organization of neurophysins, oxytocin, [8-lysine]-vasopressin and adenosine triphosphatase in porcine pituitary lobes. Biochem. J. 137, 361–371 (1973)
Robinson, A.G.: Isolation, assay and secretion of individual human neurophysins. J. clin. Invest. 55, 360–367 (1975)
Robinson, A.G., Zimmerman, E.A., Frantz, A.G.: Physiologic investigation of posterior pituitary binding proteins neurophysin-I and neurophysin-II. Metabolism 20, 1148–1155 (1971)
Sachs, H.: Vasopressin biosynthesis-I in vivo studies. J. Neurochem. 5, 297–303 (1960)
Sachs, H.: Neurosecretion. Advanc. Enzymol. 32, 327–372 (1969)
Sachs, H., Takabatake, Y.: Evidence for a precursor in vasopressin biosynthesis. Endocrinology 75, 943–948 (1964)
Skowsky, W.R., Fisher, D.A.: The use of thyroglobulin to induce antigenicity to small molecules. J. Lab. clin. Med. 80, 134–144 (1972)
Sokol, H.W., Zimmerman, E.A., Sawyer, W.H., Robinson, A.G.: The hypothalamic-neurohypophysial system of the rat. Localization and quantitation of neurophysin by light microscopic immunocytochemistry in normal rats and in Brattleboro rats deficient in vasopressin and a neurophysin. Endocrinology 98, 1176–1188
Vandesande, F., Dierickx, K., De Mey, J.: Identification of the vasopressin-neurophysin-II and the oxytocin-neurophysin-I producing neurons in the bovine hypothalamus. Cell Tiss. Res. 156, 189–200 (1975a)
Vandesande, F., Dierickx, K., De Mey, J.: Identification of separate vasopressin-neurophysin-II and oxytocin-neurophysin-I containing nerve fibres in the external region of the bovine median eminence. Cell Tiss. Res. 158, 509–516 (1975b)
Watkins, W.B.: Immunocytochemical study of the hypothalamo-neurohypophysial system I. Localization of neurosecretory neurons containing neurophysin-I and neurophysin-II in the domestic pig. Cell Tiss. Res. 175, 165–181 (1976)
Zimmerman, A.E.: Localization of hypothalamic hormones by immunocytochemical techniques. In: Frontiers in neuroendocrinology, Vol. 4, pp. 25–62 (L. Martini, W.F. Ganong, eds.). New York: Raven Press 1976
Zimmerman, E.A., Hsu, K.C., Robinson, A.G., Carmel, P.W., Frantz, A.G., Tannenbaum, M.: Studies of neurophysin secretory neurons with immunoperoxidase techniques employing antibody to bovine neurophysin. I. Light microscopic findings in monkey and bovine tissues. Endocrinology 92, 931–940 (1973)
Zimmerman, E.A., Robinson, A.G., Husain, M.K., Acosta, M., Frantz, A.G., Sawyer, W.H.: Neurohypophysial peptides in the bovine hypothalamus. The relationship of neurophysin-I to oxytocin, and neurophysin-II to vasopressin in supraoptic and paraventricular regions. Endocrinology 95, 931–936 (1974)
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This work was supported by the Medical Research Council of New Zealand
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Watkins, W.B., Choy, V.J. Immunocytochemical study of the hypothalamo-neurohypophysial system. Cell Tissue Res. 180, 491–503 (1977). https://doi.org/10.1007/BF00220170
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DOI: https://doi.org/10.1007/BF00220170