Abstract
The distribution and morphology of met-enkephalin-like immunoreactive (MetLI) and leu-enkephalin-like immunoreactive (LeuLI) neurons were investigated in the central nervous system of three gastropod species, Helix pomatia, Lymnaea stagnalis and Aplysia californica. Differences between the three species were observed in (1) the characteristics of immunostaining with antibodies to met-enkephalin and leu-enkephalin antibodies; (2) the number of immunostained neurons; (3) the projection areas of imunostained elements; (4) the specificity of immunostaining. Differences in the appearance of MetLI and LeuLI neurons were apparent: in Aplysia, both MetLI and LeuLI neurons could be observed, whereas in Lymnaea LeuLI was only found in fibers; only MetLI neurons occurred in Helix. According to an absorbtion control specificity test, a part of the LeuLI seen in the neuropil of Aplysia ganglia did not represent authentic leu-enkephalin. In Helix pomatia, significantly more MetLI neurons were present than in the CNS of Lymnaea and Aplysia; the majority of these neurons were concentrated in the cerebral ganglia and were small-size (12–25 μm) interneurons. In addition to central and peripheral projections observed in the three species, the connective tissue sheath around the ganglia and peripheral nerves contained MetLI varicose axons only in Helix, where a possible neurohormonal role could be attributed to these substance. The mapping and detailed chemical-neuroanatomical description of enkephalin-immunoreactive elements may furnish a chemical-neuroanatomical background to facilitate further neurophysiologic and pharmacologic analysis of enkephalinergic mechanisms in the gastropod CNS.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benjamin PR, Swindale NV, Slade CT (1976) Electrophysiology of identified neurosecretory neurones in the pond snail, Lymnaea stagnalis (L.). In: Salánki J (ed) Neurobiology of invertebrates: Gastropoda brain. Akadémiai Kiadó, Budapest, pp 85–100
Boer HH, Minnen J van, Stefano GB, Leung MK (1986) Opioid peptides in the central nervous system (CNS) of Lymnaea stagnalis. In: Boer HH, Geraerts WPM, Joosse J (eds) Molluscan neurobiology. North-Holland, Amsterdam Oxford New York, pp 68–72
Burrowes WR, Assanah P, Stefano GB (1983) Behavioral effects of opiates on the land snail Helix aspersa. Life Sci 33:381–384
Dalton LM, Widdowson PS (1989) The involvement of opioid peptides in stress-induced analgesia in the slug Arion ater. Peptides 10:9–13
Dyakonova TL, Moroz LL, Winlow W (1991) Facilitation of electrical coupling between identified central neurones by met-enkephalin in Helix and Lymnaea. J Physiol [Lond] 438:283P
Elekes K (1992) Neurotransmitters in the gastropod CNS: comparative immunocytochemistry. Acta Biol Hung 43:213–220
Elekes K, Nässel DR (1990) Distribution of FMR Famide-immunoreactive neurons in the central nervous system of the snail, Helix pomatia. Cell Tissue Res 262:177–190
Elekes K, Kemenes G, Hiripi L, Geffard M, Benjamin PR (1991) Dopamine-immunoreactive neurones in the central nervous system of the pond snail Lymnaea stagnalis L. J Comp Neurol 307:214–224
Guietérrez R, Asai M (1991) IR-met- and IR-leu-enkephalin content in the periesophageal ganglia of Helix aspersa. Seasonal variations. Comp Biochem Physiol [C] 100:609–613
Hernádi L, Terano Y, Muneoka Y, Kiss T (1992) Characterization of catch-relaxing peptide (CARP) immunoreactive neurons in the Helix nervous system. Acta Biol Hung 44:93–95
Jahan-Parwar B, Fredman SM (1976) Cerebral ganglion of Aplysia: cellular organization and origin of nerves. Comp Biochem Physiol 53 [A]:347–357
Joosse J (1964) Dorsal bodies and dorsal neurosecretory cells of the cerebral ganglia of Lymnaea stagnalis (L.). Arch Néerl Zool 16:1–103
Kandel ER (1976) Cellular basis of behavior. Freeman, San Francisco
Kavaliers M (1988) Novelty-induced opioid analgesia in the terrestrial snail, Capea nemoralis. Physiol Behav 42:29–32
Kavaliers M, Hirst M (1984) The presence of an opioid system mediating behavioral thermoregulation in the terrestrial snail, Cepea nemoralis. Neuropharmacology 23:1285–1289
Kavaliers M, Hirst M (1987) Slugs and snails and opiate tales: opioids and feeding behavior in invertebrates. Fed Proc 46:168–172
Kemenes G, 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 18:193–208
Kemenes G, S-Rózsa K, Stefano GB, Carpenter DO (1992) Distinct receptors for leu- and met-enkephalin on the metacerebral giant cell of Aplysia. Cell Mol Neurobiol 12:107–119
Kream RM, Zukin RS, Stefano GB (1981) Demonstration of two classes of opiate binding sites in the nervous system of the marine mollusc Mytilus edulis: positive homotropic cooperativity of lower affinity binding sites. J Biol Chem 225:9218–9224
Leung MK, Stefano GB (1984) Isolation and identification of enkephalins in pedal ganglia of Mytilus edulis (Mollusca). Proc Natl Acad Sci USA 81:955–958
Leung MK, S-Rózsa K, Hall A, Kuruvilla S, Stefano GB, Carpenter DO (1986) Enkephalin-like substance in Aplysia nervous tissue and actions of leu-enkephalin on single neurons. Life Sci 38:1529–1534
Martin R, Haas C, Voigt K-H (1986) Opioid and related neuropeptides in molluscan neurons. In: Stefano GB (ed) Handbook of comparative opioid and related neuropeptide mechanisms. vol 1. CRC Press, Boca Raton, pp 49–64
Miller MW, Alevizos A, Cropper EC, Vilim FS, Karagogeos D, Kupfermann I, Weiss KR (1991) Localization of myomodulin-like immunoreactivity in the central nervous system and peripheral tissues of Aplysia californica. J Comp Neurol 314:627–644
Nässel DR, Elekes K (1984) Ultrastructural demonstration of serotonin immunoreactivity in the nervous system of an insect (Calliphora erythrocephala). Neurosci Lett 48:203–210
Salánki J, S-Rózsa K (1986) Interaction of opioid peptides and low-molecular-weight neurotransmitters in Helix neurons. In: Stefano GB (ed) Handbook of comparative opioid and related neuropeptide mechanisms. CRC Press, Boca Ranton, pp 103–113
Schot LPC, Boer HH, Swaab DF, Van Norden S (1981) Immunocytochemical demonstration of peptidergic neurons in the central nervous system of the pond snail Lymnaea stagnalis with antisera raised to biologically active peptides of vertebrates. Cell Tissue Res 216:273–291
S-Rózsa K (1984) Pharmacology of molluscan neurons. Progr Neurobiol 23:79–150
S-Rózsa K, Solntseva EJ (1986) Modulation of cholinergic transmission by opiate peptide and FMRFamide on identified neurons of Helix pomatia (Gastropoda, Mollusca). Acta Physiol Hung 67:429–433
S-Rózsa K, Stefano GB, Salánki J, Carpenter DA (1991) Characterization of responses to enkephalins and FMRFamide on B neurons of the cerebral ganglion of Aplysia. Comp Biochem Physiol [C] 99:403–412
Stefano GB, Martin R (1983) Enkephalin-like immuno-reactivity in the pedal ganglion of Mytilus edulis (Bivalvia) and its proximity to dopamine-containing structures. Cell Tissue Res 230:147–153
Stefano GB, Vadász I, Hiripi L (1980a) Naloxone selectively blocks dopamine response of Br-type neuron in Helix pomatia L. Experientia 35:1337–1338
Stefano GB, S-Rózsa K, Hiripi L (1980b) Actions of methionine enkephalin and morphine on single neuronal activity in Helix pomatia. Comp Biochem Physiol [C] 66:193–198
Stefano GB, S-Rózsa K, Salánki J (1981) Behavioural effects of morphine on the land snail Helix pomatia. Demonstration of tolerance. Adv Physiol Sci 31:285–294
Sternberger L (1979) Immunocytochemistry. John Wiley, Chichester
Takayanagi H, Takeda N (1988) Coexistence of FMRFamide, metenkephalin and serotonin in molluscan neurons. Comp Biochem Physiol [A] 91:613–620
Tremblay JP, Schlapfer WT, Woodson PBJ, Barondes SH (1974) Morphine and related compounds evidence that they decrease available neurotransmitter in Aplysia californica. Brain Res 81:107–118
Valasek Zs, S-Rózsa K, Fedulova SA (1992) Effect of serotonin and opiate peptides on ionic-currents of dyalised Helix neurons. Acta Biol Hung 43:69–77
Waziri R (1976) Morphine effects on cholinergic synaptic transmission in Aplysia: evidence for receptor blockade. Comp Biochem Physiol [C] 55:95–102
Walker RJ (1986) Transmitters and modulators. In: Willows AOD (ed) The Mollusca, vol 8. Neurobiology and behaviour, Academic Press, Orlando, pp 279–485
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Elekes, K., Stefano, G.B. & Carpenter, D.O. Enkephalin-like immunoreactive neurons in the central nervous system of gastropods (Helix pomatia, Lymnaea stagnalis, Aplysia californica): a comparative immunocytochemical study. Cell Tissue Res 272, 329–341 (1993). https://doi.org/10.1007/BF00302738
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00302738