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In vivo and in vitro studies on the appearance of LHRH neurons in the hypothalamus of perinatal rats

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Summary

Ontogenetic development of LHRH-containing neurons was studied by fluorescence and enzyme immunohistochemistry in rats. In in vitro studies, the tissues of the septal-chiasmatic and mediobasal hypothalamic areas of fetal rats on day 16.5 or 18.5 of gestation were trypsinized separately for dissociation of the neural cells, and cultured for several days. Immunopositive reaction against LHRH was first detected in nerve cells derived from both areas of the hypothalamus of the fetuses on days 16.5 and 18.5 of gestation, after 8 and 6 days culture, respectively. The cells were small, and seemed to be bipolar in morphology indicating an axon and arborized dendrites. Immunopositive material occurred in the cell soma as well as in the cellular processes. In in vivo studies, immunopositive material, possibly deposited in nerve fibers, appeared first in OVLT and simultaneously in the external layer of the median eminence of fetuses on day 20.5 of gestation. The immunoreactive fibers increased in number in both parts with development, especially after birth in the median eminence. No immunopositive material was detected within any neural cell bodies nor in the cytoplasm of any ependymal cells.

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References

  • Araki, S., Toran-Allerand, C.D., Ferin, M., Vande Wiele, R.L.: Immunoreactive gonadotropinreleasing hormone (Gn-RH) during maturation in the rat: ontogeny of regional hypothalamic differences. Endocrinology, 97, 693–697 (1975)

    Google Scholar 

  • Aubert, M.L., Grumbach, M.M., Kaplan, S.L.: The ontogenesis of human fetal hormones. IV. Somatostatin, luteinizing hormone releasing factor, and thyrotropin releasing factor in hypothalamus and cerebral cortex of human fetuses 10–22 weeks of age. J. clin. Endocr. 44, 1130–1141 (1977)

    Google Scholar 

  • Baker, B.L., Dermody, W.C.: Effect of hypophysectomy on immunocytochemically demonstrated gonadotropin-releasing hormone in the rat brain. Endocrinology 98, 1116–1122 (1976)

    Google Scholar 

  • Barnea, A., Neaves, W.B., Porter, J.C.: Ontogeny of the subcellular compartmentalization of thyrotropin releasing hormone and luteinizing hormone releasing hormone in the rat hypothalamus. Endocrinology 100, 1068–1079 (1977)

    Google Scholar 

  • Barry, J.: Immunofluorescence study of LRF neurons in man. Cell Tiss. Res. 181, 1–14 (1977)

    Google Scholar 

  • Barry, J., Dubois, M.P.: Étude en immunofluorescence de la différenciation prénatal des cellules hypothalamiques élaboratrices de LH-RF et de la maturation de la voie neurosécrétrice préopticoinfundibulaire chez le cobaye. Brain Res. 67, 103–113 (1974)

    Google Scholar 

  • Barry, J., Dubois, M.P., Carette, B.: Immunofluorescence study of the preoptico-infundibular LRF neurosecretory pathway in the normal, castrated or testosterone-treated male guinea pig. Endocrinology 95, 1416–1423 (1974)

    Google Scholar 

  • Barry, J., Dubois, M.P., Poulain, P.: LRF producing cells of the mammalian hypothalamus. A fluorescent antibody study. Z. Zellforsch. 146, 351–366 (1973)

    Google Scholar 

  • Bassiri, R.M., Utiger, R.D.: The preparation and specificity of antibody to thyrotropin releasing hormone. Endocrinology 90, 722–727 (1972)

    Google Scholar 

  • Bugnon, C., Bloch, B., Fellmann, D.: Étude immuno-cytologique des neurones hypothalamiques à LHRH chez le foetus humain. Brain Res. 128, 249–262 (1977a)

    Google Scholar 

  • Bugnon, C., Bloch, B., Fellmann, D.: Cyto-immunological study of the ontogenesis of the gonadotropic hypothalamo-pituitary axis in the human fetus. J. Steroid Bioch. 8, 565–575 (1977b)

    Google Scholar 

  • Bugnon, C., Bloch, B., Lenys, D., Fellmann, D.: Ultrastructural study of the LH-RH containing neurons in the human fetus. Brain Res. 137, 175–180 (1977c)

    Google Scholar 

  • Daikoku, S., Kotsu, T., Hashimoto, M.: Electron microscopic observations on the development of the median eminence in perinatal rats. Z. Anat. Entwickl.-Gesch. 134, 311–326 (1971)

    Google Scholar 

  • Daikoku, S., Morishita, H., Hashimoto, T., Takahashi, A.: Light-microscopic studies on the development of the interrelationship between the neurosecretory pathway and the portal system in rats. Endocr. jap. 14, 209–224 (1967)

    Google Scholar 

  • Daikoku, S., Watanabe, Y.G.: Ultrastructural studies on the developing hypothalamo-hypophysial axis: in vivo and in vitro experiments. Gunma Symp. Endocrinol. 11, 69–85 (1974)

    Google Scholar 

  • Gross, D.S., Baker, B.L.: Immunohistochemical localization of gonadotropin-releasing hormone (GnRH) in the fetal and early postnatal mouse brain. Amer. J. Anat. 148, 195–216 (1977)

    Google Scholar 

  • Hoffman, G.E., Knigge, K.M., Moynihan, J.A., Melnyk, V., Arimura, A.: Neuronal fields containing luteinizing hormone releasing hormone (LHRH) in mouse brain. Neuroscience 3, 219–231 (1978)

    Google Scholar 

  • Ishikawa, H., Shiino, M., Rennels, E.G.: Effects of fetal brain extract on the growth and differentiation of rat pituitary anlage cells. Proc. Soc. exp. Biol. (N.Y.) 155, 511–515 (1977)

    Google Scholar 

  • Kalra, S.P.: Tissue levels of luteinizing hormone-releasing hormone in the preoptic area and hypothalamus, and serum concentrations of gonadotropins following anterior hypothalamic deafferentation and estrogen treatment of the female rat. Endocrinology 99, 101–107 (1976)

    Google Scholar 

  • Krieg, W.J.S.: The hypothalamus of the albino rat. J. comp. Neurol. 55, 19–89 (1932)

    Google Scholar 

  • Leonardelli, J.: Effects d'injections intraventriculaires de prolactine sur les neurones à LH-RH de l'hypothalamus du cobaye mâle. Etude immunocytologique. Ann. Endocr. (Paris) 37, 323–324 (1976)

    Google Scholar 

  • Paulin, C, Dubois, M.P., Barry, J., Dubois, P.M.: Immunofluorescence study of LH-RH producing cells in the human fetal hypothalamus. Cell Tiss. Res. 182, 341–345 (1977)

    Google Scholar 

  • Saito, S., Musa, K., Oshima, I., Yamamoto, S., Funato, T.: Radioimmunoassay for luteinizing hormone releasing hormone in plasma. Endocr. jap. 22, 267–273 (1975)

    Google Scholar 

  • Sétáló, G., Vigh, S., Schally, A.V., Arimura, A., Flerkó, B.: LH-RH-containing neural elements in the rat hypothalamus. Endocrinology 96, 135–142 (1975)

    Google Scholar 

  • Shiino, M., Ishikawa, H., Rennels, E.G.: In vitro and in vivo studies on cytodifferentiation of pituitary clonal cells derived from the epithelium of Rathke's pouch. Cell Tiss. Res. 181, 473–485 (1977)

    Google Scholar 

  • Silverman, A.J.: Distribution of leuteinizing hormone-releasing hormone (LHRH) in the guinea pig brain. Endocrinology 99, 30–41 (1976)

    Google Scholar 

  • Silverman, A.J., Antunes, J.L., Ferin, M., Zimmerman, E.A.: The distribution of luteinizing hormone-releasing hormone (LHRH) in the hypothalamus of the rhesus monkey. Light microscopic studies using immunoperoxidase technique. Endocrinology 101, 134–142 (1977)

    Google Scholar 

  • Sternberger, L.A., Hardy, P.H., Jr., Cuculis, J.J., Meyer, H.G.: The unlabeled antibody enzyme method of immunohistochemistry preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J. Histochem. Cytochem. 18, 315–333 (1970)

    Google Scholar 

  • Watanabe, Y.G., Daikoku, S.: Immunohistochemical study on adenohypophysial primordia in organ culture. Cell Tiss. Res. 166, 407–412 (1976)

    Google Scholar 

  • Watanabe, Y.G., Daikoku, S.: An immunohistochemical study on the cytogenesis of adenohypophysial cells in fetal rats. Develop. Biol. (in press) (1978)

  • Zimmerman, E.A., Antunes, J.L.: Organization of the hypothalamic-pituitary system: current concepts from immunohistochemical studies. J. Histochem. Cytochem. 24, 807–815 (1976)

    Google Scholar 

  • Zimmerman, E.A., Hsu, K.C., Ferin, M., Kozlowski, G.P.: Localization of gonadotropin-releasing hormone (Gn-RH) in the hypothalamus of the mouse by immunoperoxidase technique. Endocrinology 95, 1–8 (1974)

    Google Scholar 

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This work was financed by the Ministry of Education, Japan. No. 257008. We would like to thank Dr. Katsuhiko Saito (Department of Surgery, Tokushima University) for his kind advice on the preparation of the antibody used for the immunofluorescence study.

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Daikoku, S., Kawano, H., Matsumura, H. et al. In vivo and in vitro studies on the appearance of LHRH neurons in the hypothalamus of perinatal rats. Cell Tissue Res. 194, 433–445 (1978). https://doi.org/10.1007/BF00236164

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