Summary
The granules of mucosal mast cells (MMC) in the rat and man apparently poorly fixed with formaldehyde and special fixation techniques are normally used to demonstrate MMC glycosaminoglycans (GAG) in these two species. However such techniques do not permit the study of MMC granule enzyme cytochemistry or the demonstration of eosinophils. We have, therefore, examined some histochemical and immunocytochemical properties of MMC and eosinophils in normal and parasitised rats following various fixation procedures. Immersion fixation of rat intestine in 4% paraformaldehyde for 6 h not only facilitated the demonstration of MMC glycosaminoglycans with basic dyes but also permitted the concomitant staining of cosinophils with acidophilic dyes. A MMC granule-associated serine esterase was also demonstrated by enzyme cytochemistry and rat mast cell protease II was detected within MMC granules by immunocytochemistry. This new methodology obviates the requirement for separate fixation procedures in the identification and characterisation of MMC/eosinophil interactions in normal and parasitised rat intestinal mucosa.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Askenase PW (1980) Immunopathology of parasitic diseases: Involvement of basophil and mast cells. Springer Semin Immunopathol 2:417–442
Befus AD, Bienenstock J (1982) Factors involved in symbiosis and host resistance at the mucosa-parasite interface. Prog Allergy 31:76–177
Befus AD, Johnston N, Bienenstock J (1979) Nippostrongylus brasiliensis: mast cell and histamine levels in tissues of infected and normal rats. Exp Parasitol 48:1–8
Befus AD, Pearce FL, Gauldie J, Horsewood P, Bienenstock J (1982) Mucosal mast cells. I. Isolation and functional characteristics of rat intestinal mast cells. J Immunol 128:2475–2480
Crowle PK, Phillips DE (1983) Characteristics of mast cells in Chediak-Higashi mice: light and electron microscopic studies of connective tissue and mucosal mast cells. Exp Cell Biol 51:130–139
Enerback L (1966a) Mast cells in rat gastrointestinal mucosa. I. Effects of fixation. Acta Pathol Microbiol Scand 66:289–302
Enerback L (1966b) Mast cells in rat gastrointestinal mucosa. II. Dye binding and metachromatic properties. Acta Pathol Microbiol Scand 66:303–312
Enerback L (1981) The gut mucosal mast cell. Monogr Allergy 17:222–232
Encrback L, Kolset SO, Kusche M, Hjerpe A, Lindahl U (1984) Glycosaminotlycans in rat mucosal mast cells. (Submitted for publication)
Graham RC, Karnovsky MJ (1966) The early stages of absorbtion of injceted horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291
Graham RC, Lundholm U, Karnovsky MJ (1965) Cytochemical demonstration of peroxidase activity with 3-amino-9-ethyl carbazole. J Histochem Cytochem 13:150
Heyderman E, Neville AM (1977) A shorter immunoperoxidase technique for the demonstration of carcinoembryonic antigen and other cell products. J Clin Pathol 30:138
Huntley JF, Newlands G, Miller HRP (1984) The isolation and characterisation of globule leucocytes: their derivation from mucosal mast cells in parasitised sheep. Parasite Immunol 6:371–390
Jarrett EEE, Heig DM (1984) Mucosal mast cells in vivo and in vitro. Immunol Today 5:115–118
Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27:137a
Lendrum AC (1944) The staining of eosinophil polymorphs and enterochromaffin cells in histological sections. J Pathol Bacteriol 56:441
Miller HRP (1971) Immune reactions in mucous membranes. II. The differentiation of intestinal mast cells during helminth expulsion in the rat. Lab Invest 24:339–347
Miller HRP, Huntley JF, Dawson A, McL (1981) Mucus secretion in the gut, its relationship to the immune response in Nippostrongylus-infected rats. In: Bourne FJ (ed) Current topics in veterinary medicine and animal science. Martinus and Nijhoff, London The Hague, pp 402–430
Miller HRP, Jarrett WFH (1971) Immune reactions in mucous membranes. I. Intestinal mast cell response during helminth expulsion in the rat. Immunology 20:277
Miller HRP, Walshaw R (1972) Immune reactions in mucous membranes. IV. Histochemistry of intestinal mast cells during helminth expulsion in the rat. Am J Pathol 69:195–206
Nawa Y, Miller HRP (1978) Protection aganist Nippostrongylus brasiliensis by adonptive immunization with immune thoracic duct lymphocytes. Cell Immunology 37:51–60
Pearse AGE (1968) Chemistry of fixation in Histochemistry theoretical and applied. JA Churchill, London, pp 70–73
Ruitenberg EJ, Gustowska L, Elgersma A, Ruitenberg HM (1982) Effect of fixation on the light microscopical visualisation of mast cells in the mucosa and connective tissue of the human duodenum. Int Arch Allergy Appl Immunol 67:233–238
Strobel S, Miller HRP, Ferguson A (1981) Human intestinal mucosal mast cells: Evaluation of fixation and staining techniques. J Clin Pathol 34:851–858
Tas J, Berndsen RG (1977) Does heparin occur in mucosal mast cells of the rat small intestine? J Histochem Cytochem 25:1058–1062
Wingren U, Enerback L (1983) Mucosal mast cells of the rat intestine: A re-evaluation of fixation and staining properties, with special reference to protein blocking and solubility of the granular glycosaminoglycans. Histochem J 15:571–582
Woodbury RG, Gruzenski GM, Lagunoff D (1978) Immunofluorescent localization of a serine protease in rat small intestine. Proc Natl Acad Sci USA 75:2785–2789
Woodbury RG, Miller HRP (1982) Quantitative analysis of mucosal mast cell protease in the intestines of Nippostrongylus-infected rats. Immunology 46:487–495
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Newlands, G.F.J., Huntley, J.F. & Miller, H.R.P. Concomitant detection of mucosal mast cells and eosinophils in the intestines of normal and nippostrongylus-immune rats. Histochemistry 81, 585–589 (1984). https://doi.org/10.1007/BF00489539
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00489539