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Formation of crystalloid inclusions in the small intestine of neonatal pigs: an immunocytochemical study using colloidal gold

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Summary

Enterocytes of the small intestine in 1-day-old suckling piglets contain numerous vesicles in the apical cytoplasm and a large granule located beneath the nucleus. Within the next 3 days, these granules transform into electron-dense crystalloid inclusions. These membrane-bound inclusions are up to 10 μm in length and 1–2 μm in diameter, and they are composed of electron-dense lamellae 3.9 nm apart. Postembedding immunocytochemistry, using rabbit anti-porcine IgG and goat anti-rabbit IgG conjugated to 10 nm colloidal gold, revealed that both the granules and the crystalloid inclusions contained a high concentration of maternal IgG. Although the IgG content of the crystalloid inclusions was detected on epoxy-embedded sections, the use of LR White resin resulted in a much higher density of labelling. Quantification of the labelling density showed that the concentration of IgG in the crystalloid inclusions was approximately ten times higher than that in the lumenal colostrum and approximately three times higher than that in the granules. These observations showed that there are at least three compartments involved in the accretion of IgG in the small intestine of neonatal piglets: smaller apical endocytotic vesicles, large subnuclear granules and crystalloid inclusions. The role of these compartments in maternal immunoglobulin absorption and in the acquisition of passive immunity has yet to be explored.

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References

  • Baintner, K. (1986) Intestinal Absorption of Macromolecules and Immune Transmission from Mother to Young. Boca Raton, Florida: CRC Press.

    Google Scholar 

  • Beesley, J. E. & Betts, M. P. (1986) A quantitative investigation into the role of buffer components for reducing non-specific immunolabelling. Histochem. J. 18, 55.

    Google Scholar 

  • Bendayan, M. & Zollinger, M. (1983) Ultrastructural localization of antigenic sits on osmium-fixed tissues applying the protein A-gold technique. J. Histochem. Cytochem. 31, 101–09.

    Google Scholar 

  • Bendayan, M., Nanci, A. & Kan, F. W. K. (1987) Effect of tissue processing on colloidal gold cytochemistry. J. Histochem. Cytochem. 35, 983–96.

    Google Scholar 

  • Berryman, M. A. & Rodewald, R. D. (1990) An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes. J. Histochem. Cytochem. 38, 159–70.

    Google Scholar 

  • Birrell, G. B., Hedberg, K. K. & Griffith, O. H. (1987) Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy. J. Histochem. Cytochem. 35, 843–53.

    Google Scholar 

  • Clarke, R. M. & Hardy, R. N. (1971) Histological changes in the small intestine of the young pig and their relation to macromolecular uptake. J. Anat. 108, 63–77.

    Google Scholar 

  • Craig, S. & Goodchild, D. J. (1982) Post-embedding immunolabeling. Some effects of tissue preparation on the antigenicity of plant proteins. Eur. J. Cell Biol. 28, 251–56.

    Google Scholar 

  • DelCerro, M., Cogen, J. & Del Cerro, C. (1980) Stevenel's blue, an excellent stain for optical microscopical study of plastic embedded tissues. Microsc. Acta 83, 117–21.

    Google Scholar 

  • Desjardins, M. & Bendayan, M. (1989) Ultrastructural distribution of endogenous IgGs in the glomerular wall of control and diabetic rats. Histochem. J. 21, 731–42.

    Google Scholar 

  • Dixon, W. J. (1990) BMDP Biomedical Computer Programs. Berkeley, CA: University of California Press.

    Google Scholar 

  • Erickson, P. A., Anderson, D. H. & Fisher, S. K. (1987) Use of uranyl acetate en bloc to improve tissue preservation and labeling for post-embedding immunoelectron microscopy. J. Elect. Microsc. Tech. 5, 303–14.

    Google Scholar 

  • Fujita, M., Reinhart, F. & Neutra, M. (1990) Convergence of apical and basolateral endocytic pathways at apical late endosomes in absorptive cells of suckling rat ileum in vivo. J. Cell Sci. 97, 385–94.

    Google Scholar 

  • Gabriel, L., Escribano, O., Perales, J., Bellas, C., Odriozola, J. & Navarro, J. L. (1985) Multiple myeloma with crystalline inclusions in most hemopoietic cells. Amer. J. Hematol. 18, 405–11.

    Google Scholar 

  • Hardy, R. N., Hockaday, A. R. & Tapp, R. L. (1971) Observations on the structure of the small intestine in foetal, neonatal and suckling pigs. Philos. Trans. Royal Soc. London B259, 517–31.

    Google Scholar 

  • Hasegawa, H., Nakamura, A., Watanabe, H., Brown, W. R. & Nagura, H. (1987) Intestinal uptake of IgG in suckling rats. Distinction between jejunal and ileal epithelial cells demonstrated by simultaneous ultrastructural localization of IgG and acid phosphatase. Gastroenterology 92, 186–91.

    Google Scholar 

  • Hearn, S. A., Silver, M. M. & Sholdice, J. A. (1985) Immunoelectron microscopic labeling of immunoglobulin in plasma cells after osmium fixation and epoxy embedding. J. Histochem. Cytochem. 33, 1212–18.

    Google Scholar 

  • Hurez, D., Flandrin, G., Preud'homme, J. L. & Seligman, N. M. (1972) Unreleased intracellular monoclonal macroglobulin in chronic lymphocytic leukaemia. Clin. Exp. Immunol. 10, 223–34.

    Google Scholar 

  • Jennette, J. C., Wilkman, A. S. & Benson, J. D. (1981) IgD myeloma with intracytoplasmic crystalline inclusions Am. J. Clin. Pathol. 75, 231–35.

    Google Scholar 

  • Johnson, A. B. & Bettica, A. (1989) On-grid immunogold labeling of glial intermediate filaments in epoxyembedded tissue. Amer. J. Anat. 185, 335–41.

    Google Scholar 

  • Kadota, K. & Niibori, S. (1985) A case of swine follicular lymphoma lymphoma with intracytoplasmic immunoglobulin inclusions. J. Comp. Pathol. 95, 599–608.

    Google Scholar 

  • Klobasa, F., Butler, J. E. (1987) Absolute and relative concentrations of immunoglobulins G, M, and A, and albumin in the lacteal secretion of sows of different lactation numbers. Am. J. Vet. Res. 48, 176–82.

    Google Scholar 

  • Klobasa, F., Werhahn, E. & Butler, J. E. (1987) Composition of sow milk during lactation. J. Anim. Sci. 64, 1458–66.

    Google Scholar 

  • Kőműves, L. G., Turner, D. S., Mckee, K. S., Nichols, B. L. & Heath, J. P. (1990) An immunocytochemical study of maternal immunoglobulin localization in the suckling pig intestine. Proc. XIIth. Int. Congr. Electron Microscopy 3, 922–23.

    Google Scholar 

  • Kramarcy, N. R. & Sealock, R. (1991) Commercial preparations of colloidal gold-antibody complexes frequently contain free active antibody. J. Histochem. Cytochem. 39, 37–9.

    Google Scholar 

  • Murata, H. & Namioka, S. (1977) The duration of colostral immunoglobulin uptake by the epithelium of the small intestine of neonatal piglets. J. Comp. Pathol. 87, 431–9.

    Google Scholar 

  • Newman, G. R. (1987) Use and abuse of LR White. Histochem. J. 19, 118–20.

    Google Scholar 

  • Newman, J., Jasani, B. & Williams, E. D. (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope. Histochem. J. 15, 543–55.

    Google Scholar 

  • Oliver, C., Auth, R. E. & Hand, A. R. (1979) Formation and fate of ethionine-induced cytoplasmic crystalloids in rat parotid acinar cells. Am. J. Anal. 155, 185–200.

    Google Scholar 

  • Pabst, R. (1987) The anatomical basis for the immune function of the gut. Anat. Embryol. 176, 135–44.

    Google Scholar 

  • Rodewald, R. (1973) Intestinal transport of antibodies in the newborn rat. J. Cell Biol. 58, 189–211.

    Google Scholar 

  • Roth, J., Bendayan, M. & Orci, L. (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J. Histochem. Cytochem. 26, 1074–81.

    Google Scholar 

  • Schvartz, H., Bonhomme, P., Caulet, S., Beorchia, A., Patey, M. & Caulet, J. (1985) Bone marrow lambda-type light chain crystalline structures associated with multiple myeloma. Virchows Arch. 407, 449–56.

    Google Scholar 

  • Schweitzer, P. P. A., Taylor, S. E. & Schultz, L. D. (1991) Synthesis of abnormal immunoglobulins by hybridomas from autoimmune ‘viable motheaten’ mutant mice. J. Cell Biol. 114, 35–43.

    Google Scholar 

  • Shultz, L. D., Coman, D. R., Bailey, C. L., Beamer, W. G. & Sidman, C. L. (1984) ‘Viable motheaten’, a new allele at the motheaten locus. I. Pathology. Amer. J. Pathol. 116, 179–92.

    Google Scholar 

  • Schultz, L. D., Coman, D. R., Lyons, B. L., Sidman, C. L. & Taylor, S. (1987) Development of plasmacytoid cells with Russell bodies in autoimmune ‘viable motheaten’ mice. Amer. J. Pathol. 127, 38–50.

    Google Scholar 

  • Staley, T. E., Jones, E. W. & Corley, L. D. (1969a) Crystals in the intestinal epithelium of neonatal pigs. Pathol. Vet. 6, 454–62.

    Google Scholar 

  • Staley, T. E., Jones, E. W. & Corley, L. D. (1969b) Fine structure of duodenal absortive cells in the newborn pig before and after feeding of colostrum. Amer. J. Vet. Res. 30, 567–81.

    Google Scholar 

  • Széky, A., Rátz, F., Tuboly, S. & Nagy, F. (1979) Absorption of colostral immunoglobulins in suckling piglets. Acta Microbiol. Acad. Sci. Hung. 26, 99–110.

    Google Scholar 

  • Viale, G., Dell'orto, P., Braidotti, P. & Coggi, G. (1985) Ultra-structural localization of intracellular immunoglobulins in Epon-embedded human lymph nodes. J. Histochem. Cytochem. 33, 400–6.

    Google Scholar 

  • Yamamoto, T., Hishida, A., Honda, N., Ito, I., Shirasawa, H. & Nagase, M. (1991) Crystal-storing histiocytosis and crystalline tissue deposition in multiple myeloma. Arch. Pathol. Lab. Med. 115, 351–4.

    Google Scholar 

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Authors and Affiliations

  1. Microscopy Laboratory, USDA/ARS Children's Nutrition Research Center, Baylor College of Medicine, 77030-2600, Houston, Texas, USA

    Lázló G. Kőműves, Buford L. Nichols, T. William Hutchens & Julian P. Heath

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  1. Lázló G. Kőműves
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  2. Buford L. Nichols
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  3. T. William Hutchens
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  4. Julian P. Heath
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Kőműves, L.G., Nichols, B.L., Hutchens, T.W. et al. Formation of crystalloid inclusions in the small intestine of neonatal pigs: an immunocytochemical study using colloidal gold. Histochem J 25, 19–29 (1993). https://doi.org/10.1007/BF00161041

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  • DOI: https://doi.org/10.1007/BF00161041

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