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

Systematic presence of GABA-immunoreactivity in the tubero-infundibular and tubero-hypophyseal dopaminergic axonal systems: an ultrastructural immunogold study on several mammals

  • 42 Accesses

  • 30 Citations


Immunoreactivities for tyrosine hydroxylase (TH), gamma-aminobutyric acid (GABA) and, in some cases, glutamic acid decarboxylase (GAD) were detected by light and electron microscopy in axons projecting into the median eminence and pituitary gland of various mammals (rats, mice, guinea pigs, cats, rabbits and hares). Light microscope immunoperoxidase reactions were performed on adjacent semithin sections of plasticembedded samples. In the median eminence external zone, the distributions of the TH- and GAD- or GABA-immunoreactive endings were very similar in the anterior and lateral areas, while medially the GABA-labelled endings predominated. Comparable distribution patterns were found in the various species examined. In the pituitary gland, the distributions of GABA- and TH- immunoreactivities were superimposable in the intermediate lobes of all species examined, except in the rabbit and hare in which both types of innervation were lacking. For electron microscopy, the immunogold procedure was applied to sections of lowicryl-embedded samples; simultaneous detection of GABA- and TH-immunoreactivities was enabled by recto-verso double labelling with gold particles of distinct diameters. In the median eminence, GABA-immunoreactivity occurred systematically in the TH-positive endings, while distinct GABA-positive/TH-negative axons were also detected. In the intermediate lobe, the colocalization of TH- and GABA-immunoreactivities was a constant feature of the axons innervating the melanotrophic cells in all the species examined, except in the Leporidae. The functional significance of this colocalization remains to be determined.

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


  1. Anderson R, Mitchell R (1985) Effects of GABA receptor agonists on (3H)dopamine release from median eminence and pituitary neurointermediate lobe. Eur J Pharmacol 115:109–112

  2. Anderson R, Mitchell R (1986) Uptake and autoreceptor-controlled release of (3H) GABA by the hypothalamic median eminence and pituitary neurointermediate lobe. Neuroendocrinology 42:277–284

  3. Bargmann W, Lindner E, Andres KH (1967) Über Synapsen an endokrinen Epithelzellen und die Definition sekretorischer Neurone: Untersuchungen am Zwischenlappen der Katzenhypophyse. Z Zellforsch 77:282–298

  4. Bartfai T, Iverfeldt K, Fisone G (1988) Regulation of the release of coexisting neurotransmitters. Ann Rev Pharmacol Toxicol 28:285–310

  5. Bartfai T, Bertorelli R, Consolo S, Diaz-Arnesto L, Fisone G, Hökfelt T, Iverfeldt K, nPalazzi E, Ogren SO (1988/1989) Acute and chronic studies on functional aspects of coexistence. J Physiol 83:126–132

  6. Ben-Jonathan N, Arbogast LA, Hyde JF (1989) Neuroendocrine regulation of prolactin release. Progr Neurobiol 33:399–447

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

  8. Brownstein JM, Palkovits M, Tappaz ML, Saavedra M, Kizer JS (1976) Effect of surgical isolation of the hypothalamus on its neurotransmitter content. Brain Res 117:287–295

  9. Dahlström A, Fuxe K (1964) Evidence for the existence of monoamine-containing neurons in the central nervous system. I. Demonstration of monoamines in the cell bodies of brain stem neurons. Acta Physiol Scand Suppl 232:1–55

  10. Dahlström A, Fuxe K (1966) Monoamines and the pituitary gland. Acta Endocrinol 51:301–314

  11. Decavel C (1987) Le système hypothalamo-posthypophysaire: étude immunocytochimique de différentes composantes de son innervation et de leurs interrelations avec les neurones neurosécrétoires chez le Rat. Doctoral thesis. Université Bordeaux I, France

  12. Demeneix BA, Feltz P, Loeffler JP (1986) GABAergic mechanisms and their functional relevance in the pituitary. In: Erdö SL, Bowery NG (eds) GABAergic mechanisms in the mammalian periphery. Raven Press, New York, pp 261–289

  13. Eberle A (1988) The melanotropins. Chemistry, physiology and mechanisms of action. Karger, Basel

  14. Enjalbert A, Ruberg M, Arancibia S, Fiore L, Priam M, Kordon C (1979) Independent inhibition of prolactin secretion by dopamine and gamma-aminobutyric acid in vitro. Endocrinology 105:823–826

  15. Everitt BJ, Hökfelt T, Wu JY, Goldstein M (1984) Coexistence of tyrosine hydroxylase-like and gamma-aminobutyric acid-like immunoreactivities in neurons of the arcuate nucleus. Neuroendocrinology 39:189–191

  16. Everitt BJ, Meister B, Hökfelt T, Melander T, Terenius L, Rökaeus A, Theodorsson-Norheim E, Dockray G, Edwardson J, Cuello AC, Elde R, Goldstein M, Hemmings H, Ouimet C, Walaas I, Greengard P, Vale W, Weber E, Wu JY (1986) The hypothalamic arcuate nucleus-median eminence complex: immunohistochemistry of transmitters, peptides and DARPP-32 with special reference to coexistence in dopamine neurons. Brain Res Rev 11:97–155

  17. Felman K (1990) Rôle du GABA dans les régulations neuroendocrines. Les systèmes GABAergiques tubéro-infundibulaires: recherche de rétro-contrôles endocrines. Doctoral thesis. Université Claude Bernard, Lyon I, France

  18. Felman K, Tappaz ML (1989) GABAergic biochemical parameters of the tubero-infundibular neurons following chronic hyperprolactinemia. Neuroendocrinology 49:580–585

  19. Felman K, Tappaz ML (1990) Evidence for a short feedback of prolactin on the tubero-infundibular endings: differential effect on the release of (3H) gamma-aminobutyric acid and (3H) dopamine from superfused median eminence. J Neuroendocrinology (in press)

  20. Fuxe K, Hökfelt T (1966) Further evidence for the existence of tubero-infundibular dopamine neurons. Acta Physiol Scand 66:243–244

  21. Hökfelt T, Johansson O, Goldstein M (1984) Chemical anatomy of the brain. Science 225:1326–1334

  22. Kosaka T, Kosaka K, Hataguchi Y, Nagatsu I, Wu JY, Ottersen OP, Storm-Mathisen J, Hama K (1987) Catecholaminergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat. Exp Brain Res 66:191–210

  23. Locatelli V, Apud JA, Gudelsky GA, Cocchi D, Masotto C, Casanueva F, Racagni G, Muller EE (1985) Prolactin in cerebrospinal fluid increases the synthesis and release of hypothalamic gamma-aminobutyric acid. J Endocrinol 106:323–328

  24. Lundberg JM, Rudehill A, Sollevi A, Theodorsson-Norheim E, Hamberger B (1986) Frequency- and reserpine-dependent chemical coding of sympathetic transmission: differential release of noradrenaline and neuropeptide Y from pig spleen. Neurosci Lett 63:96–100

  25. Meister B, Hökfelt T (1988) Peptide- and transmitter-containing neurons in the mediobasal hypothalamus and their relation to GABAergic systems: possible roles in control of prolactin and growth hormone secretion. Synapse 2:585–605

  26. Meister B, Hökfelt T, Steinbusch HWM, Skagerberg G, Lindvall O, Geffard M, Joh TH, Cuello AC, Goldstein M (1988) Do tyrosine hydroxylase-immunoreactive neurons in the ventrolateral arcuate nucleus produce dopamine or only L-DOPA? J Chem Neuroanat 1:59–64

  27. Oertel WH, Mugnaini E, Tappaz ML, Weise VK, Dahl AL, Schmechel DE, Kopin IJ (1982) Central GABAergic innervation of neurointermediate pituitary lobe: biochemical and immunocytochemical study in the rat. Proc Natl Acad Sci USA 79:675–679

  28. Okamura H, Kitahama K, Raynaud B, Nagatsu I, Borri-Voltatorni C, Weber M (1988a) Aromatic L-amino acid decarboxylase (AADC)-immunoreactive cells in the tuberal region of the rat hypothalamus. Biomed Res 9:261–267

  29. Okamura H, Kitahama K, Mons N, Ibata Y, Jouvet M, Geffard M (1988b) L-DOPA-immunoreactive neurons in the rat hypothalamic tuberal region. Neurosci Lett 95:42–46

  30. Okamura H, Kitahama K, Nagatsu I, Geffard M (1988e) Comparative topography of dopamine- and tyrosine hydroxylase-immunoreactive neurons in the rat arcuate nucleus. Neurosci Lett 95:347–353

  31. O'Malley D, Masland RH (1989) Co-release of acetylcholine and γ-aminobutyric acid by a retinal neuron. Proc Natl Acad Sci USA 86:3414–3418

  32. Pickel VM, Hyub Joh T, Field PM, Becker CG, Reis DJ (1975) Cellular localization of tyrosine hydroxylase by immunohistochemistry. J Histochem Cytochem 23:1–12

  33. Plantjé JF, Schipper J, Piet FHM, Verheijden PFHM, Stoof JC (1987) D2-dopamine receptors regulate the release of (3H)dopamine in rat basal hypothalamus and neurointermediate lobe of the pituitary gland. Brain Res 413:205–212

  34. Racké K, Grosshans A, Sirrenberg S, Ziegler K (1988) Presynaptic regulation of the electrically evoked release of endogenous dopamine from isolated neurointermediate lobe or isolated neural lobe of the rat pituitary gland in vitro. NaunynSchmiedeberg's Arch Pharmacol 337:504–511

  35. Sakaue M, Saito N, Taniguchi H, Baba S, Tanaka C (1988) Immunohistochemical localization of gamma-aminobutyric acid in the rat pituitary gland and related hypothalamic regions. Brain Res 44:343–353

  36. Saland LC, Wallace JA, Samora A, Gutierrez L (1988) Co-localization of tyrosine hydroxylase (TH)- and serotonin (5-HT)- immunoreactive innervation in the rat pituitary gland. Neurosci Lett 94:39–45

  37. Sarkar DK, Gottschall PE, Meites J, Horn A, Dow RC, Fink G, Cuello AC (1983) Uptake and release of (3H)dopamine by the median eminence: evidence for presynaptic dopaminergic receptors and for dopaminergic feedback inhibition. Neuroscience 10:821–830

  38. Schimchowitsch S, Stoeckel ME, Klein MJ, Garaud JC, Schmitt G, Porte A (1983) Oxytocin-immunoreactive nerve fibers in the pars intermedia of the pituitary in the rabbit and hare. Cell Tis Res 228:255–263

  39. Schimchowitsch S, Palacios JM, Stoeckel ME, Schmitt G, Porte A (1986) Absence of inhibitory dopaminergic control of the rabbit pituitary gland intermediate lobe. Neuroendocrinology 42:71–74

  40. Schimchowitsch S, Vuillez P, Klein MJ, Tappaz ML, Stoeckel ME (1988) GABA colocalizes with dopamine in the axons projecting in the pituitary neuro-intermediate lobe and median eminence in various species. In: Europ J Neurosci, Suppl, Abstr 11th Ann Meet Europ Neurosci Ass, p 11

  41. Stoeckel ME, Schimchowitsch S, Garaud JC, Schmitt G, Vaudry H, Klein MJ, Porte A (1985a) Immunocytochemical evidence for intragranular processing of pro-opiomelanocortin in the melanotropic cells of the rabbit. Cell Tis Res 242:365–370

  42. Stoeckel ME, Tappaz ML, Hindelang C, Seweryn C, Porte A (1985b) Opposite effects of monosodium glutamate on the dopaminergic and GABAergic innervations of the median eminence and the intermediate lobe in the mouse. Neurosci Lett 56:249–255

  43. Tappaz ML, Aguera M, Belin MF, Pujol JF (1980) Autoradiography of GABA in the rat hypothalamic median eminence. Brain Res 186:379–391

  44. Tappaz ML, Brownstein MJ (1977) Origin of glutamate decarboxylase (GAD)-containing cells in discrete hypothalamic nuclei. Brain Res 132:95–106

  45. Tappaz ML, Kakucska I, Paut L, Makara B (1986) Decreased GABAergic innervation of the pituitary intermediate lobe after rostral hypothalamic cuts. Brain Res Bull 17:711–717

  46. Tappaz ML, Wassef M, Oertel WH, Paut L, Pujol JF (1983) Light and electron-microscopic immunocytochemistry of glutamic acid decarboxylase (GAD) in the basal hypothalamus. Morphological evidence for neuroendocrine gamma-aminobutyrate (GABA). Neuroscience 9:271–287

  47. Valentino KL, Crumrine DA, Reichardt LF (1985) Lowicryl K4M embedding of brain tissue for immunogold electron microscopy. J Histochem Cytochem 33:969–973

  48. Vincent SR, Hökfelt T, Wu JY (1982) GABA neuron systems in the hypothalamus and the pituitary gland. Neuroendocrinology 34:117–125

  49. Vuillez P, Carbajo-Pérez S, Stoeckel ME (1987) Colocalization of GABA and tyrosine hydroxylase immunoreactivities in the axons innervating the neurointermediate lobe of the rat pituitary: an ultrastructural immunogold study. Neurosci Lett 79:53–58

  50. Walaas I, Fonnum F (1978) The effect of parenteral glutamate treatment on the localization of neurotransmitters in the mediobasal hypothalamus. Brain Res 153:549–562

Download references

Author information

Correspondence to S. Schimchowitsch.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schimchowitsch, S., Vuillez, P., Tappaz, M.L. et al. Systematic presence of GABA-immunoreactivity in the tubero-infundibular and tubero-hypophyseal dopaminergic axonal systems: an ultrastructural immunogold study on several mammals. Exp Brain Res 83, 575–586 (1991).

Download citation

Key words

  • Intermediate lobe
  • Median eminence
  • GABA
  • Dopamine
  • Colocalization
  • Mammal