Experimental Brain Research

, Volume 85, Issue 1, pp 1–9 | Cite as

Localization of dopamine and its relation to the growth hormone producing cells in the central nervous system of the snail Lymnaea stagnalis

  • T. R. Werkman
  • J. van Minnen
  • P. Voorn
  • H. W. M. Steinbusch
  • B. H. C. Westerink
  • T. A. De Vlieger
  • J. C. Stoof


The distribution of dopamine in the central nervous system of the pond snail Lymnaea stagnalis was investigated by using immunocytochemistry and HPLC measurements. With both methods it was demonstrated that dopamine is predominantly present in the cerebral and pedal ganglia. The dopamine-immunoreactivity was mainly observed in nerve-fibers in the neuropile of the ganglia. Relatively few dopamine-immunopositive cell bodies (diameters 10–30 μm) were found. A large cell in the right pedal ganglion (the so-called RPeD1) stained positively with the dopamine antibody. It has previously been demonstrated that the growth hormone producing cells (GHCs) possess dopamine receptors on their cell bodies. However, dopamine-immunopositive fibers were observed only in the vicinity of the GHC nerve-endings and not close to the GHC cell bodies.

Key words

Immunocytochemistry Dopamine Growth hormone producing cells Snail 


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  1. Audesirk G (1985) Amine-containing neurons in the brain of Lymnaea stagnalis: distribution and effects of precursors. Comp Biochem Physiol 81A:359–365Google Scholar
  2. Boer HH, Schot LPC, Steinbusch HWM, Montagne C, Reichelt D (1984) Co-existence of immunoreactivity to anti-dopamine, anti-serotonin and anti-vasotocin in the cerebral giant neuron of the pond snail Lymnaea stagnalis. Cell Tissue Res 238:411–412Google Scholar
  3. Buijs RM, Geffard M, Pool CW, Hoorneman EMD (1984) The dopaminergic innervation of the supraoptic and paraventricular nucleus: a light and electron microscopical study. Brain Res 323:65–72Google Scholar
  4. Cottrel GA, Abernethy KB, Barrand MA (1979) Large aminecontaining neurones in the central ganglia of Lymnaea stagnalis. Neuroscience 4:685–689Google Scholar
  5. Croll RP (1988) Distribution of monoamines within the central nervous system of the juvenile pulmonate snail, Achatina fulica. Brain Res 460:29–49Google Scholar
  6. Croll RP, Chiasson BJ (1989) Postembryonic development of serotoninlike immunoreactivity in the central nervous system of the snail Lymnaea stagnalis. J Comp Neurol 280:122–142Google Scholar
  7. Dahlström A, Fuxe K (1964) Evidence for the existence of monoamine containing neurons in the central nervous system. I. Demonstration of monoamines in cell bodies of brain neurons. Acta Physiol Scand 62 Suppl 232:1–55Google Scholar
  8. De Biasi S, Vitellaro-Zuccarello L, Blum I (1984) Histochemical localization of monoamines and cholinesterases in Mytilus pedal ganglion. Histochemistry 81:561–565Google Scholar
  9. De Vlieger TA, Lodder JC, Stoof JC, Werkman TR (1986) Dopamine receptor stimulation induces a potassium dependent hyperpolarizing response in growth hormone producing cells of the gastropod mollusc Lymnaea stagnalis. Comp Biochem Physiol 83C:429–433Google Scholar
  10. Elekes K, Hustert R, Geffard M (1987) Serotonin-immunoreactive and dopamine-immunoreactive neurones in the terminal ganglion of the cricket, Acheta domestica: light- and electron-microscopic immunocytochemistry. Cell Tissue Res 250:167–180Google Scholar
  11. Falck B, Hillarp HA, Thieme G, Thorp A (1962) Fluorescence of catecholamines and related compounds condensed with formaldehyde. J Histochem Cytochem 10:348–354Google Scholar
  12. Geffard M, Buijs RM, Seguela P, Pool CW, Le Moal M (1984) First demonstration of highly specific and sensitive antibodies against dopamine. Brain Res 294:161–165Google Scholar
  13. Kemenes GY, Elekes K, Hiripi L, Benjamin PR (1989) A comparison of four techniques for mapping the distribution of serotonin and serotonin-containing neurons in fixed and living ganglia of the snail Lymnaea. J Neurocytol 18:193–208Google Scholar
  14. Klemm N (1983) Monoamine-containing neurons and their projections in the brain (supraoesophageal ganglion) of cockroaches: an aldehyde fluorescence study. Cell Tiss Res 229:379–402Google Scholar
  15. Klemm N, Nassel DR, Osborne NN (1985) Dopamine-beta-hydroxylase-like immunoreactive neurons in two insect species, Calliphora erythrocephala and Periplaneta americana. Histochemistry 83:159–164Google Scholar
  16. Konings PNM, Vullings HGB, Geffard M, Buijs RM, Diederen JHB, Jansen WF (1988) Immunocytochemical demonstration of octopamine-immunoreactive cells in the nervous system of Locusta migratoria and Schistocerca gregaria. Cell Tissue Res 251:371–379Google Scholar
  17. McCaman MW, Ono JK, McCaman RE (1979) Dopamine measurements in molluscan ganglia and neurons using a new, sensitive assay. J Neurochem 32:1111–1113Google Scholar
  18. McCaman MW, Ono JK, McCaman RE (1984) 5-Hydroxytryptamine measurements in molluscan ganglia and neurons using a modified radioenzymatic assay. J Neurochem 43:91–99Google Scholar
  19. Mefford IN, Foutz A, Noyce N, Jurik SM, Handen C, Dement WC, Barchas JD (1982) Distribution of norepinephrine, epinephrine, dopamine, serotonin, 3,4-dihydroxyphenylacetic acid, homovanilic acid and 5-hydroxyindole-3-acetic acid in dog brain. Brain Res 236:339–349Google Scholar
  20. Mons N, Geffard M (1987) Specific antisera against catecholamines: L-3,4-dihydroxyphenylalanine, dopamine, noradrenaline, and octopamine tested by an enzyme-linked immunosorbent assay. J Neurochem 48:1826–1833Google Scholar
  21. Pickel VM, Joh TH, Reis D (1976) Monoamines synthesizing enzymes in central dopaminergic, noradrenergic and serotonergic neurons: immunohistochemical localization by light and electron microscopy. J Histochem Cytochem 24:792–806Google Scholar
  22. Slade CT, Mills J, Winlow W (1981) The neuronal organization of the paired pedal ganglia of Lymnaea stagnalis (L.). Comp Biochem Physiol 69A:789–803Google Scholar
  23. Smit AB, Vreugdenhil E, Ebberink RHM, Geraerts WPM, Klootwijk J, Joosse J (1988) Growth-controlling molluscan neurons produce the precursor of an insulin-related peptide. Nature (Lond) 331:535–538Google Scholar
  24. Steinbusch HWM, Bol GJM, Karlsson L, De Vente J (1991) Immunocytochemistry of dopamine: comparison of formaldehyde versus glutaraldehyde fixation. Histochemistry (in press)Google Scholar
  25. Steinbusch HWM, Tilders JH (1987) Immunohistochemical techniques for light-microscopical localization of dopamine, noradrenaline, adrenaline, serotonin and histamine in the central nervous system. In: Steinbusch HWM (ed) Monoaminergic neurons: light microscopy and ultrastructure. John Wiley, Chichester, pp 125–166Google Scholar
  26. Steinbusch HWM, Verhofstad AAJ, Joosten HWF (1978) Localization of serotonin in the central nervous system by immunocytochemistry: description of a specific and selective technique and some applications. Neuroscience 3:811–819Google Scholar
  27. Steinbusch HWM, Verhofstad AAJ, Joosten HWJ (1983) Antibodies to serotonin: methodological aspects and applications. In: Cuello AC (eds) Immunohistochemistry. IBRO handbook series, Vol. 3. John Wiley, Chichester, pp 193–214Google Scholar
  28. Sternberger LA (1986) Immunocytochemistry. John Wiley & Sons, New York Chichester Brisbane Toronto, pp 90–200Google Scholar
  29. Stoof JC, De Vlieger TA, Lodder JC (1984) Opposing roles for D-l and D-2 dopamine receptors in regulating the excitability of growth-hormone producing cells in the snail Lymnaea stagnalis. Eur J Pharmacol 106:431–435Google Scholar
  30. Stoof JC, Werkman TR, Lodder JC, De Vlieger TA (1986) Growth hormone producing cells in the snail Lymnaea stagnalis as a model system for mammalian dopamine receptors? Trends Pharmacol Sci 7:7–9Google Scholar
  31. Takeda S, Vieillemaringe J, Geffard M, Remy C (1986) Immunohistological evidence of dopamine cells in the cephalic nervous system of the silkworm Bombyx mori: coexistence of dopamine and endorphin-like substance in neurosecretory cells of the suboesophageal ganglion. Cell Tiss Res 243:125–128Google Scholar
  32. Van Minnen J, Boer HH (1987) Generation and application of monoclonal antibodies raised against homogenates of whole central nervous systems of the pond snail Lymnaea stagnalis. Proceedings Kon Ned Akad Wetenschappen Serie C 90:193–201Google Scholar
  33. Van Minnen J, Schallig HDFH (1990) Demonstration of insulinrelated substances in the central nervous systems of pulmonates and Aplysia californica. Cell Tiss Res 260:381–386Google Scholar
  34. Vieillemaringe J, Duris P, Geffard M, Le Moal M, Delaage M, Bensch C, Girardie J (1984) Immunohistochemical localization of dopamine in the brain of the insect Locusta migratoria migratorides in comparison with the catecholamine distribution determined by the histofluorescence technique. Cell Tiss Res 237:391–394Google Scholar
  35. Voorn P, Kalsbeek A, Jorritsma-Byham B, Groenewegen HJ (1988) The pre- and postnatal development of the dopaminergic cell groups in the ventral mesencephalon and the dopaminergic innervation of the striatum of the rat. Neuroscience 25:857–888Google Scholar
  36. Werkman TR (1989) Dopaminergic neurotransmission in the central nervous system of Lymnaea stagnalis. Ph. D. thesis. Free University Press, AmsterdamGoogle Scholar
  37. Werkman TR, De Vlieger TA, Stoof JC (1990) Indications for a hormonal function of dopamine in the central nervous system of the snail Lymnaea stagnalis. Neurosci Lett 108:167–172Google Scholar
  38. Werkman TR, De Vlieger TA, Van Minnen J, Voorn P, Stoof JC (1988) Dopamine mediated regulation of the neuroendocrine growth hormone cells of the snail Lymnaea stagnalis. In: Dahlströhm A, Belmaker RH, Sandler M (eds) Progress in catecholamine research, Part A. Basic aspects and peripheral mechanisms, neurology and neurobiology, Vol 42A. Alan R Liss Inc, New York, pp 325–329Google Scholar
  39. Werkman TR, Lodder JC, De Vlieger TA, Stoof JC (1987) Further pharmacological characterization of a D-2-like dopamine receptor on growth hormone producing cells in Lymnaea stagnalis. Eur J Pharmacol 139:155–161Google Scholar
  40. Westerink BHC, Mulder TBA (1981) Determination of picomole amounts of dopamine, noradrenaline, 3,4-dihydroxyphenylalanine, 3,4-dihydroxy-phenyl-acetic acid, homovanillic acid, and 5-hydroxyindolacetic acid in nervous tissue after one step purification on sephadex G-10, using high-performance liquid chromatography with a novel type of electrochemical detection. J Neurochem 36:1449–1462Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • T. R. Werkman
    • 1
  • J. van Minnen
    • 4
  • P. Voorn
    • 2
  • H. W. M. Steinbusch
    • 3
  • B. H. C. Westerink
    • 5
  • T. A. De Vlieger
    • 4
  • J. C. Stoof
    • 1
  1. 1.Department of Neurology, Medical FacultyFree UniversityBT AmsterdamThe Netherlands
  2. 2.Department of Anatomy, Medical FacultyFree UniversityBT AmsterdamThe Netherlands
  3. 3.Department of Pharmacology, Medical FacultyFree UniversityBT AmsterdamThe Netherlands
  4. 4.Department of Neurophysiology, Biological FacultyFree UniversityHV AmsterdamThe Netherlands
  5. 5.Pharmaceutical Laboratory, University of GroningenGroningenThe Netherlands

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