Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Monoaminergic tracts of the diencephalon and innervation of the pars intermedia in Rana temporaria

A fluorescence and microspectrofluorimetric study

  • 35 Accesses

  • 46 Citations


With the fluorescence technique of Falck-Hillarp two monoaminergic tracts having independent nuclear sources, and extending towards the hypophysis, were identified in the diencephalon of Rana temporaria. The nature of the fluorophore in the diencephalic nuclei which give rise to the tracts, and also that of the nerve fibers innervating the pars intermedia (PI), were analyzed microspectrofluorimetrically.

The first tract, the preoptic recess organ (PRO)-hypophysial tract, arises from the neurons of the PRO, traverses the preoptic region, ascends towards the dorsal chiasmatic area, curves down and extends posteriorly along the mid-ventral region of the tuber cinereum towards the median eminence. Apparently this pathway has no contact with either the paraventricular organ (PVO) or the nucleus infundibularis dorsalis (NID). The second pathway, the paraventricular organ (PVO)-hypophysial tract, arises mainly from the PVO of each side, traverses the tuber cinereum and converges posteriorly to join the PRO-hypophysial tract at the hind end of the tuber cinereum. Participation of the NID neurons in the formation of this tract could not be excluded. It is argued that the PVO-hypophysial tract, but not the PRO-hypophysial tract which runs along the mid-ventral region of the tuber cinereum, may be responsible for transportation of the monoamines from the PVO/NID to the pars intermedia.

Microspectrofluorimetric study indicated that the PRO neurons contain only dopamine, whereas two types of neuronal fluorophores were observed in the PVO and NID. Many perikarya in both these nuclear sites possess dopamine, while some contain 5-hydroxytryptamine (5HT) and/or 5-hydroxytryptophane (5HTP). An analysis of the fluorescent nerve fibers in the PI revealed a double innervation. The first category includes dopaminergic fibers, whereas the second type seems to be adrenergic or noradrenergic in nature.

On leave from the Department of Zoology, Nagpur University, Nagpur, India.

This is a preview of subscription content, log in to check access.


  1. Bargmann, W.: Weitere Untersuchungen am neurosekretorischen Zwischenhirn-Hypophysensystem. Z. Zellforsch. 42, 247–272 (1955)

  2. Bartels, W.: Die Ontogenese der aminhaltigen Neuronensysteme im Gehirn von Rana temporaria. Z. Zellforsch. 116, 94–118 (1971)

  3. Baumgarten, H. G.: Biogenic monoamines in the cyclostome and lower vertebrate brain. Progr. Histochem. Cytochem. 4, 1–90 (1972)

  4. Baumgarten, H. G., Braak, H.: Catecholamine im Hypothalamus vom Goldfisch (Carassius auratus). Z. Zellforsch. 80, 246–263 (1967)

  5. Björklund, A., Ehinger, B., Falck, B.: A method for differentiating dopamine from noradrenaline in tissue sections by microspectrofluorimetry. J. Histochem. Cytochem. 16, 263–270 (1968)

  6. Björklund, A., Ehinger, B., Falck, B.: Analysis of fluorescence excitation peak ratios for the cellular identification of noradrenaline, dopamine or their mixtures. J. Histochem. Cytochem. 20, 56–64 (1972a)

  7. Björklund, A., Falck, B., Owman, C.: Fluorescence microscopic and microspectrofluorimetric techniques for the cellular localization and characterization of biogenic amines. In: S. Barson (ed.), Methods of investigative and diagnostic endocrinology, vol. 1, The thyroid and biogenic amines, ed. by J. E. Rall and I. J. Kopin. Amsterdam: North-Holland Publishing Co. 1972b

  8. Björklund, A., Falck, B., Stenevi, U.: Microspectrofluorimetric characterization of monoamines in the central nervous system; evidence for a new neuronal monoamine-like compound. Progr. Brain Res. 34, 63–73 (1971)

  9. Björklund, A., Moore, R. Y., Nobin, A., Stenevi, U.: The organization of tubero-hypophysial and reticulo-infundibular catecholamine neuron systems in the rat. Brain Res. 51, 171–191 (1973)

  10. Braak, W.: Biogene Amine im Gehirn vom Frosch (Rana esculenta). Z. Zellforsch. 106, 269–308 (1970)

  11. Calas, A., Hartwig, H.-G., Collin, J. P.: Noradrenergic innervation of the median eminence. Microspectrofluorimetric and pharmacological study in the duck, Anas platyrhynchos. Cell Tiss. Res. 147, 491–504 (1974)

  12. Dawson, D. C., Ralph, C. L.: Neural control of the amphibian pars intermedia: Electrical responses evoked by illumination of the lateral eyes. Gen. comp. Endocr. 16, 611–614 (1971)

  13. Dierickx, K., Druyts, A., Vandenberghe, M. P., Goossens, N.: Identification of adenohypophysiotropic neurohormone producing neurosecretory cells in Rana temporaria. I. Ultrastructural evidence for the presence of neurosecretory cells in the tuber cinereum. Z. Zellforsch. 134, 459–504 (1972)

  14. Dierickx, K., Goossens, N., Vandenberghe, M. P.: Identification of adenohypophysiotropic neurohormone producing neurosecretory cells in Rana temporaria. III. The tubero-hypophysial monoaminergic fibres and the role of the tubero-hypophysial neurosecretory system. Z. Zellforsch. 143, 93–106 (1973)

  15. Doerr-Schott, J., Follenius, E.: Localisation des fibres aminergiques dans l'hypophyse de Rana esculenta. Etude autoradiographique au microscope électronique. C. R. Acad. Sci. (Paris) 269, 737–740 (1969)

  16. Doerr-Schott, J., Follenius, E.: Innervation de l'hypophyse intermédiaire de Rana esculenta, et identification des fibres aminergiques par autoradiographie au microscope électronique. Z. Zellforsch. 106, 99–118 (1970)

  17. Enemar, A., Falck, B.: On the presence of adrenergic nerves in the pars intermedia of the frog, Rana temporaria. Gen. comp. Endocr. 5, 577–583 (1965)

  18. Enemar, A., Falck, B., Iturriza, F. C.: Adrenergic nerves in the pars intermedia of the pituitary in the toad, Bufo arenarum. Z. Zellforsch. 77, 325–330 (1967)

  19. Falck, B., Hillarp, N. Å., Thieme, G., Torp, A.: Fluorescence of catecholamines and related compounds condensed with formaldehyde. J. Histochem. Cytochem. 10, 348–354 (1962)

  20. Fuxe, K., Hökfelt, T.: The influence of central catecholamine neurons on the hormone secretion from the anterior and posterior pituitary. In: Neurosecretion (ed. Stutinsky, F.), p. 165–177. Berlin-Heidelberg-New York: Springer 1967

  21. Fuxe, K., Hökfelt, T.: Participation of central monoamine neurons in the regulation of anterior pituitary function with special regard to the neuroendocrine role of tuberoinfundibular dopamine neurons. In: Aspects of neuroendocrinology (eds. Bargmann, W., Scharrer, B.), p. 192–205. Berlin-Heidelberg-New York: Springer 1970

  22. Fuxe, K., Hökfelt, T., Nilsson, O.: Effect of constant light and androgen-sterilization on the amine turnover of the tubero-infundibular dopamine neurons: blockade of cyclic activity and induction of a persistent high dopamine turnover in the median eminence. Acta endocr. (Kbh.) 69, 635–639 (1972)

  23. Goos, H.J.Th.: Hypothalamic control of the pars intermedia in Xenopus laevis tadpoles. Z. Zellforsch. 97, 118–124 (1969)

  24. Goossens, N., Dierickx, K., De Waele, G.: The vascularization and monoaminergic structures of the organon vasculosum laminae terminalis of Rana temporaria. Z. Zellforsch. 143 527–534 (1973)

  25. Herrick, C. D.: The brain of the tiger salamander Ambystoma tigrinum. Chicago-London: Chigaco Univ. Press 1948

  26. Hogben, L. T., Slome, D.: The pigmentary effectory system. VI. The dual character of endocrine co-ordination in amphibian colour change. Proc. roy. Soc. B 108, 10–53 (1931)

  27. Hökfelt, T., Fuxe, K.: On the morphology and the neuroendocrine role of the hypothalamic catecholamine neurons. In: Brain-endocrine interaction. Median eminence: Structure and function (eds. Knigge, K. M., Scott, D. E., Weindl, A.) p. 181–223. Basel: S. Karger 1972

  28. Hopkins, C. R.: Localization of adrenergic fibers in the amphibian pars intermedia by electron microscope autoradiography and their selective removal by 6-hydroxydopamine. Gen. comp. Endocr. 16, 112–120 (1971)

  29. Ito, T.: Changes in skin colour and fine structure of the intermediate pituitary gland of the frog, Rana nigromaculata, after extirpation of the median eminence. Neuroendocrinology 8, 180–197 (1971)

  30. Iturriza, F. C.: Electron-microscopic study of the pars intermedia of the pituitary of the toad Bufo arenarum. Gen. comp. Endocr. 4, 492–502 (1964)

  31. Jørgensen, C. B.: Central nervous control of adenohypophysial functions. In: Perspectives in endocrinology. Hormones in the lives of vertebrates (eds. Barrington, E. J. W., Jørgensen, C. B.), p. 469–541. London: Academic Press 1968

  32. Kemali, M., Braitenberg, V.: Atlas of the frog's brain. Berlin-Heidelberg-New York: Springer 1969

  33. McKenna, O. C., Rosenbluth, J.: Cytological evidence for secretory and sensory catecholamine-containing cell types bordering the third ventricle of the toad brain. In: VIth international symposium on neurosecretion, London, p. 38 (Abstracts) 1973

  34. Nakai, Y., Gorbman, A.: Evidence for a doubly innervated secretory unit in the anuran pars intermedia. II. Electron microscopic studies. Gen. comp. Endocr. 13, 108–116 (1969)

  35. Oordt, P.G.W.J. van, Goos, H.J.Th., Peute, J., Terlou, M.: Hypothalamo-hypophysial relations in amphibian larvae. Gen. comp. Endocr. Suppl. 3, 41–50 (1972)

  36. Oshima, K., Gorbman, A.: Evidence for a doubly innervated secretory unit in the anuran pars intermedia. I. Electrophysiologic studies. Gen. comp. Endocr. 13, 98–107 (1969)

  37. Parent, A.: Distribution of monoamine-containing neurons in the brain stem of the frog, Rana temporaria. J. Morph. 193, 67–78 (1973)

  38. Parker, G. H.: Animal colour changes and their neurohumors. Cambridge: Cambridge University Press 1948

  39. Pehlemann, F. W.: Ultrastructure and innervation of the pars intermedia of the pituitary of Xenopus laevis. Gen. comp. Endocr. 9, 481 (Abstract) (1967)

  40. Ritzén, M.: Cytochemical identification and quantitation of biogenic amines. A microspectrofluorimetric and autoradiographic study. M. D. Thesis. Institute for Medical Cell Research and Genetics, Karolinska Institutet, Stockholm, 1967

  41. Saland, L. C.: Ultrastructure of the frog pars intermedia in relation to hypothalamic control of hormone release. Neuroendocrinology, 3, 72–88 (1968)

  42. Sharp, P. J., Follett, B. K.: The adrenergic supply within the avian hypothalamus. In: Aspects of neuroendocrinology (Bargmann, W. and Scharrer, B., eds.), p. 95–103. BerlinHeidelberg-New York: Springer 1970

  43. Soest Warren, S., Farner, D. S., Oksche, A.: Fluorescence microscopy of neurons containing primary catecholamines in the ventral hypothalamus of the White-crowned Sparrow, Zonotrichia leucophrys gambelii. Z. Zellforsch. 141, 1–17 (1973)

  44. Terlou, M., Ploemacher, R. E.: The distribution of monoamines in the tel-, di- and mesencephalon of Xenopus laevis tadpoles, with special reference to the hypothalamo-hypophysial system. Z. Zellforsch. 137, 521–540 (1973)

  45. Vigh, B.: Das Paraventrikularorgan und das Zirkumventrikuläre System des Gehirns. Studia Biologica Hungaricae, Akadémiai Kiadó, Verlag der Ungarischen Akademie der Wissenschaften, Budapest, Bd. 10, S. 1–149 (1971)

  46. Vigh-Teichmann, I., Röhlich, P., Vigh, B.: Licht- und elektronenmikroskopische Untersuchungen am Recesus praeopticus-Organ von Amphibien. Z. Zellforsch. 98, 217–232 (1969 b)

  47. Vigh-Teichmann, I., Vigh, B.: Liquor-contacting neuronal areas in the periventricular gray substance of the central nervous system. Gen. comp. Endocr. 13, 537 (Abstract) (1969)

  48. Vigh-Teichmann, I., Vigh, B., Aros, B.: Fluorescence histochemical studies on the preoptic recess organ in various vertebrates. Acta biol. Acad. Sci. hung. 20, 423–436 (1969a)

  49. Vullings, H. G. B., Kers, J.: The optic tracts of Rana temporaria and a possible retino-preoptic pathway. Z. Zellforsch. 139, 179–200 (1973)

  50. Wilson, J. F., Dodd, J. M.: Distribution of monoamines in the diencephalon and pituitary of the dogfish, Scyliorhinus canicula L. Z. Zellforsch. 137, 451–469 (1973)

  51. Zambrano, D., de Robertis, E.: Ultrastructure of the peptidergic and monoaminergic neurons in the hypothalamic neurosecretory system of anuran batracians. Z. Zellforsch. 90, 230–244 (1968)

Download references

Author information

Additional information

Supported by fellowships of the Deutscher Akademischer Austauschdienst (1972/73) and the Alexander von Humboldt-Stiftung (1973/74), Federal Republic of Germany.

Supported by a grant (Ok 1/16) from the Deutsche Forschungsgemeinschaft to Professor A. Oksche.

The authors are indebted to Professor A. Oksche for his interest in this study. They also wish to thank Dr. M. Ueck and Dr. W. Möller for sharing their technical experience, Dr. R. Snipes and Mr. A. Soames for reading the English text, Miss D. Vaihinger for the drawings and Mr. W. Kramer for his technical assistance.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Prasada Rao, P.D., Hartwig, H.G. Monoaminergic tracts of the diencephalon and innervation of the pars intermedia in Rana temporaria . Cell Tissue Res. 151, 1–26 (1974).

Download citation

Key words

  • Diencephalon
  • Monoaminergic tracts
  • Pars intermedia, Rana temporaria
  • Microspectrofluorimetry