Summary
In the central nervous system of the pond snail Lymnaea stagnalis a large number of elements (cells and fibers) can be identified with antisera (a-FM) to the molluscan cardioactive tetrapeptide FMRFamide (Phe-Met-Arg-Phe-NH2). Of these elements some are also reactive to antivasotocin (a-VT) and/or anti-gastrin (a-Gas). These observations suggest that the a-FM positive elements belong to more than one type. Previous results had already indicated that the immunoreactivity of many a-FM positive cells is influenced by the type of fixation. Taking into account the effects of three fixatives on the reactivity of the cells, and their staining characteristics with the two other antisera used, 8 a-FM positive types could be distinguished.
Homologous and heterologous adsorptions were carried out to test the specificity of a-FM, a-VT and a-Gas. After homologous adsorptions no staining was obtained. After heterologous adsorptions only part of the multiple staining cells were identified. This indicates that in a-FM, a-VT and a-Gas in addition to (more) selective IgG molecules, less specific IgG molecules occur that can bind to other peptides than those used to raise the antisera (cross-reaction). The (more) selective IgG molecules in a-FM bind to 6 of the a-FM positive types, suggesting that in L. stagnalis a family of FMRFamide-like substances occurs. This conclusion is sustained by results obtained with a-FM adsorbed with fragments of FMRFamide. It appeared that the less selective IgG molecules in a-FM, a-VT and a-Gas cause the multiple stainings of those cells that remain unstained with an antiserum adsorbed with a heterologous antigen. Multiple staining, which can not be abolished by heterologous adsorption, probably is due to the binding of (more) selective IgG molecules to different antigenic determinants present in the cells.
The results show that unexpected cross-reactions may occur in immunocytochemical staining procedures. It thus seems precarious to draw conclusions about the chemical structure of a peptide solely on the basis of immunocytochemical studies.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boer HH, Schot LPC (1983) Phylogenetic aspects of peptidergic systems. In: Lever J, Boer HH (eds) Molluscan neuro-endocrinology. Monogr R Neth Acad Sci. North-Holland, Amsterdam, pp 9–14
Boer HH, Schot LPC, Roubos EW, Maat A ter, Lodder JC, Reichelt D, Swaab DF (1979) ACTH-like immunoreactivity in two electrotonically coupled giant neurons in the pond snail Lymnaea stagnalis. Cell Tissue Res 202:231–240
Boer HH, Schot LPC, Veenstra JA, Reichelt D (1980) Immunocytochemical identification of neural elements in the central nervous system of a snail, some insects, a fish and a mammal with an antiserum to the molluscan cardio-excitatory tetrapeptide FMRFamide. Cell Tissue Res 213:21–27
Dockray GJ, Vaillant C, Williams RC (1981) New vertebrate braingut peptide related to a molluscan neuropeptide and an opioid peptide. Nature 292:656–657
Duve H, Thorpe A (1979) Immunofluorescent localization of insulin-like material in the median neurosecretory cells of the blowfly Calliphora vomitoria (Diptera). Cell Tissue Res 200:187–191
Duve H, Thorpe A (1980a) Isolation and localization of an insect insulin-like material: immunological, biological and physical characteristics. Gen Comp Endocrinol 40:363–364
Duve H, Thorpe A (1980b) Localization of pancreatic polypeptide (PP)-like material in neurons of the brain of the blowfly Calliphora erythrocephala (Diptera). Cell Tissue Res 210:101–109
Geraerts WPM, Leeuwen JPThM van, Nuyt K, With ND de (1981) Cardioactive peptides of the CNS of the pulmonate snail Lymnaea stagnalis. Experientia 37:1168–1169
Grimmelikhuijzen CJP, Dockray GJ, Yanaihara N (1981) Bombesin-like immunoreactivity in the nervous system of Hydra. Histochemistry 73:171–180
Grimmelikhuijzen CJP, Dockray GJ, Schot LPC (1982) FMRFamide-like immunoreactivity in the nervous system of Hydra. Histochemistry 73:499–508
Hökfelt T, Johansson O, Ljungdal Å, Lundberg JM, Schulzberg M (1980) Peptidergic neurones. Nature 284:515–521
Martin R, Voigt KH (1981) Enkephalins co-exist with oxytocin and vasopressin in nerve terminals of rat neurohypophysis. Nature 289:502–504
Osborne NN, Dockray GJ (1982) Bombesin-like immunoreactivity in specific neurons of the snail Helix aspersa and an example of the coexistence of substance P and serotonin in an invertebrate neurone. Neurochem Int 4:175–180
Price DA (1982) The FMRFamide-like peptide of Helix aspersa. Comp Biochem Physiol 72:325–328
Price DA, Greenberg MJ (1977) Purification and characterization of a cardio-excitatory neuropeptide from the central ganglia of a bivalve mollusc. Prep Biochem 7:50–62
Roth KE, Weber E, Barchas JD (1983) Immunoreactive dynorphin (1–8) and corticotrophin releasing factor in subpopulations of hypothalamic neurons. Science 219:189–191
Schot LPC, Boer HH (1982) Immunocytochemical demonstration of peptidergic cells in the pond snail Lymnaea stagnalis with an antiserum to the molluscan cardioactive tetrapeptide FMRFamide. Cell Tissue Res 225:347–354
Schot LPC, Boer HH, Swaab DF, Noorden S van (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
Schot LPC, Boer HH, Wijdenes J (1983) Localization of neurons innervating the heart of Lymnaea stagnalis studied immunocytochemically with anti-FMRF-amide and anti-vasotocin. In: Lever J, Boer HH (eds) Molluscan neuro-endocrinology. Monogr R Neth Acad Arts Sci. North-Holland, Amsterdam, pp 203–208
Strambi C, Strambi A, Cupo A, Rougon-Rapuzzi G, Martin N (1978) Etude des taux d'une substance apparentée à la vasopresine dans le système nerveux de grillons soumis a différentes conditions hygrométriques. C R Acad Sci Paris D 287:1227–1230
Strambi C, Rougon-Rapuzzi G, Cupo A, Martin N, Strambi A (1979) Mise en évidence immunocytologique d'un composé apparentée à la vasopressine dans le système nerveux du grillon Acheta domesticus. C R Acad Sci Paris D 288:131–133
Swaab DF, Nijveldt F, Pool CW (1975) Distributions of oxytoxin and vasopressin in the rat supraoptic and paraventricular nucleus. J Endocrinol 67:461–462
Swaab DF, Pool CW, Leeuwen FW van (1977) Can specificity ever be proved in immunocytochemical staining? J Histochem Cytochem 25:388–391
Toubeau G, Desclin J, Parmentier M, Pasteels JL (1979) Compared localizations of prolactin-like and adrenocorticotrophin immunoreactivities within the brain of the rat. Neuroendocrinology 29:374–384
Vincent SR, Skirboll L, Hökfelt T, Johansson O, Lundberg JM, Elde RP, Terenius L, Kimmel J (1982) Coexistence of somatostatin-and avian pancreatic polypeptide (APP)-like immunore-activity in some forebrain neurons. Neuroscience 7:439–446
Weber E, Evans CJ, Samuelson SJ, Barchas JD (1981) Novel peptide neuronal system in rat brain and pituitary. Science 214:1248–1251
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schot, L.P.C., Boer, H.H. & Montagne-Wajer, C. Characterisation of multiple immunoreactive neurons in the central nervous system of the pond snail Lymnaea stagnalis with different fixatives and antisera adsorbed with the homologous and the heterologous antigens. Histochemistry 81, 373–378 (1984). https://doi.org/10.1007/BF00514332
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00514332