Summary
The encapsulation of experimentally introduced material was studied in Lymnaea stagnalis. Abiotic (yvelon sponge) and biotic materials (autografts, allografts and xenografts) were implanted into the cephalopedal blood sinus.
Within 24 hrs the yvelon sponge is completely invaded and encapsulated by round amoebocytes. Amoebocytes are the only type of blood cell in L. stagnalis. The number of capsule amoebocytes increases until 3 days after implantation. These capsule amoebocytes are normal blood amoebocytes. This inclusion is based on morphological observations as well as on the results of labelling experiments (blood amoebocytes were labelled with India ink or 3H-thymidine). These data are in accordance with evidence that amoebocytes may have more than one function.
From 24 hrs onwards a progressive flattening of the capsule amoebocytes was observed. In 7-day-old capsules 3 strata were distinguished: inner and outer strata consisting of tightly packed, flattened amoebocytes, and a middle stratum still composed of round amoebocytes. The cells of the outer stratum possess an extensive granular endoplasmic reticulum. During the further process of encapsulation the cells of the inner stratum become loaded with lysosomes. The middle stratum disappears: the cells flatten and become part of the other two strata. In the outer stratum, collagenous connective tissue fibrils and ground substance are deposited between the cells. This situation is reached about 1 month after implantation. Capsule formation is completed by that time: during the next 5 months, no major structural changes occur in the capsules.
The reactions to biotic materials appeared to be quite different, and show that L. stagnalis is able to discriminate between different types of graft. Autografts and allografts are not encapsulated—only a transient amoebocyte reaction occurs at the cut surfaces of the transplants—whereas xenografts evoke a severe host reaction: they are infiltrated and encapsulated by amoebocytes. It is suggested that the discrimination mechanism is located in the plasma membrane of the amoebocytes.
The encapsulation of xenografts ends in a capsule consisting only of the collagenous connective tissue stratum. The cells of the inner stratum infiltrate the graft and phagocytose degenerated graft tissue.
Autoradiographic experiments using 3H-proline as a marker for collagen synthesis showed that the flattened cells of the outer stratum produce collagenous connective tissue fibrils, indicating that these cells are fibroblasts. In experiments using ink or 3H-thymidine labelled amoebocytes it was shown that these fibroblasts are transformed amoebocytes. Finally it is suggested that amoebocytes also have the capacity to transform into myofibroblasts.
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Abolinš-Krogis, A.: Fluorescence and histochemical studies on the calcification-initiating lipofuscin type pigment granules in the shell-repair membrane of the snail, Helix pomatia L. Z. Zellforsch. 142, 205–221 (1973)
Abramoff, P., La Via, M. F.: Biology of immune responses. McGraw-Hill Book Company, New York. 1970
Allgöwer, M., Hulliger, L.: Origin of fibroblasts from mononuclear blood cells: A study on in vitro formation of the collagen precursor, hydroxyproline, in buffy coat cultures. Surgery 47, 603–110 (1960)
Bellanti, J. A.: Immunology. Philadelphia-London-Toronto: W. B. Saunders Company 1971
Brown, A. C., Brown, R. J.: The fate of thorium dioxide injected into the pedal sinus of Bullia (Gastropoda: Prosobranchiata). J. exp. Biol. 42, 509–519 (1965)
Cheney, D. P.: A summary of invertebrate leucocyte morphology with emypasis on blood elements of the manilla clam, Tapes semidecussata. Biol. Bull. 140, 353–368 (1971)
Cheng, T. C., Galloway, P. C.: Transplantation immunity in mollusks: The histoincompatibility of Helisoma duryi normale with allografts and xenografts. J. Invert. Path. 15, 177–192 (1970)
Cheng, T. C., Rifkin, E.: Cellular reactions in marine molluscs in response to helminth parasitism. A. symposium on diseases of fishes and shellfishes. Amer. Fish. Soc. spec. publ. 5, 443–496 (1970)
Cheng, T. C., Thakur, A. S., Rifkin, E.: Phagocytosis as an internal defense mechanism in the mollusca: with an experimental study of the role of leucocytes in the removal of ink particles in Littorina scabra Linn. Proc. Symp. Mollusca II, 546–563 (1969)
Cooper, E. L.: Transplantation immunity in helminths and annelids. Transpl. Proc. II, 216–221 (1970)
Daems, W. Th., Wisse, E., Brederoo, P.: Electron microscopy of the vacuolar apparatus. In: Lysosomes in biology and pathology, vol. I, p. 64–112. Eds.: Dingle, J. T., Fell, H. B. Amsterdam-London: North-Holland Publ. Comp. 1969
Davies, P. S., Partridge, T.: Limpet haemocytes. I. Studies on aggregation and spike formation. J. Cell Sci. 11, 757–769 (1972)
Faust, E. C.: Pathological changes in the gastropod liver produced by fluke infection. Bull. Johns Hopk. Hosp. 31, 79–84 (1920)
Feng, S. Y., Feng, J. S., Burke, C. N., Khairallah, L. H.: Light and electron microscopy of the leucocytes of Crassostrea virginica (Mollusca: Pelecypoda). Z. Zellforsch. 120, 222–245 (1971)
Grimstone, A. V., Rotheram, S., Salt, G.: An electron-microscope study of capsule formation by insect blood cells. J. Cell Sci. 2, 281–292 (1967)
Joosse, J., Lever, J.: Techniques of narcotization and operation for experiments with Lymnaea stagnalis (Gastropoda, Pulmonata). Proc. kon. ned. Akad. Wet. C 62, 145–149 (1959)
Kouri, J., Ancheto, O.: Transformation of macrophages into fibroblasts. Exptl. Cell Res. 71, 168–176 (1972)
McKay, D., Jenkin, C. R., Rowley, D.: Immunity in invertebrates I. Studies on the naturally occurring haemaglutinins in the fluid from invertebrates. Aust. J. exp. Biol. med. Sci. 47, 125–134 (1969)
Meuleman, E. A.: Host-parasite interrelationships between the freshwater pulmonate Biomphalaria pfeifferi and the trematode Schistosoma mansoni, Neth. J. Zool. 22, 355–427 (1972)
Nappi, A. J.: Hemocytic changes associated with the encapsulation and melanization of some insect parasites. Exp. Parasit. 33, 285–302 (1973)
Narain, A. S.: The amoebocytes of lamellibranch molluscs, with special reference to the circulating amoebocytes. Malacol. Rev. 6, 1–12 (1973)
Oehmichen, M.: Demonstration of hematogenous origin of fibroblasts by parabiosis. Experientia (Basel) 29, 841–842 (1973)
Pan, C.: Studies on the host-parasite relationship between Schistosoma mansoni and the snail Australorbis glabratus. Amer. J. trop. Med. Hyg. 14, 931–976 (1965)
Pauley, G. B., Krassner, S. M., Chapman, F. A.: Bacterial clearance in the California sea hare, Aplysia californica. J. Invert. Path. 18, 227–239 (1971)
Pauley, G. B., Sparks, A. K.: The acute inflammatory reaction in two different tissues of the pacific oyster, Crassostrea gigas. J. Fish. Res. Bd. Canada 23, 1913–1921 (1966)
Pearse, A. G. E.: Histochemistry. Theoretical and applied. London: J. & A. Churchill 1968
Prowse, R. H., Tait, N. N.: In vitro phagocytosis by amoebocytes from the haemolymph of Helix aspersa (Müller) I. Evidence for opsonic factor(s) in serum. Immunology 17, 437–443 (1969)
Rayner, C. R. W.: The source of fibroblasts and mesothelial cells appearing on an implanted prosthetic material. Proc. Wound Healing symp. Rotterdam, The Netherlands, 1974 (in press)
Reaven, E. P., Axline, S. G.: Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages. J. Cell Biol. 59, 12–27 (1973)
Ross, R., Everett, N. B., Tyler, R.: Wound healing and collagen formation VI. The origin of the wound fibroblast studied in parabiosis. J. Cell Biol. 44, 645–654 (1970)
Roubos, E. W.: Regulation of neurosecretory activity in the freshwater pulmonate Lymnaea stagnalis (L.). A quantitative electron microscopical study. Z. Zellforsch. 146, 177–205 (1973)
Ruddell, C. L.: The fine structure of the granular amoebocytes of the pacific oyster Crassostrea gigas. J. Invert. Path. 18, 269–275 (1971)
Salt, G.: The cellular defence reactions of insects. Cambridge: University Press. 1970
Stang-Voss, C.: Zur Ultrastruktur der Blutzellen wirbelloser Tiere III. Über die Haemozyten der Schnecke Lymnea stagnalis (Pulmonata). Z. Zellforsch. 107, 141–156 (1970)
Sminia, T.: Structure and function of blood and connective tissue cells of the fresh water pulmonate Lymnaea stagnalis studied by electron microscopy and enzyme histochemistry. Z. Zellforsch. 130, 497–526 (1972)
Sminia, T.: Haematopoiesis in the freshwater snail Lymnaea stagnalis studied by electron microscopy and autoradiography. Cell Tiss. Res. 150, 443–454 (1974a)
Sminia, T.: Wound healing in invertebrates, with special reference to the freshwater mollusc Lymnaea stagnalis (L.) Proc. Wound Healing symp. Rotterdam, The Netherlands, 1974b (in press)
Sminia, T., Pietersma, K., Scheerboom, J. E. M.: Histological and ultrastructural observations on wound healing in the freshwater pulmonate Lymnaea stagnalis. Z. Zellforsch. 141, 561–573 (1973)
Smith, R. T., Good, R. A.: Cellular recognition. Amsterdam: North Holland Publ. Compagny 1969
Tripp, M. R.: The fate of foreign materials experimentally introduced into the snail Australorbis glabratus. J. Parasit. 47, 745–751 (1961)
Tripp, M, R.: Defense mechanisms of mollusks. J. reticuloendoth. Soc. 7, 173–182 (1970)
Vostal, Z.: A contribution to the cytology of haemocytes of molluscs (Gastropoda). Biológia Brat. 24, 384–392 (1969)
Weiss, L.: The cells and tissues of the immune system. Structure, functions, interactions. London: Prentice-Hall Int. Inc. 1972
Wessels, N. K., Spooner, B. S., Ash, J. F., Bradley, M. O., Luduena, M. A., Taylor, E. L., Wrenn, J. T., Yamada, K. M.: Microfilaments in cellular and developmental processes. Science 171, 135–143 (1971)
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The authors wish to thank Dr. H. H. Boer for his guidance and valuable criticism during the preparation of the manuscript, Prof. Dr. J. Lever for reading the manuscript, Miss José van 't Rood for her technical assistance, Miss Benita Plesch for correcting the English text and Mrs. Anneke Teuling-Graatsma for typing the manuscript.
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Sminia, T., Borghart-Reinders, E. & van de Linde, A.W. Encapsulation of foreign materials experimentally introduced into the freshwater snail Lymnaea stagnalis . Cell Tissue Res. 153, 307–326 (1974). https://doi.org/10.1007/BF00229161
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DOI: https://doi.org/10.1007/BF00229161