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Quantitative cytochemical demonstration of intracellular thyroglobulin in cultured human thyrocytes. Effects of fixatives, TSH and Interleukin-1β

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Summary

A quantitative immunocytochemical method is described for measuring intracellular thyroglobulin in human thyrocytes grown in monolayer, based on the imidazole-enhanced 3,3′-diaminobenzidine/peroxidase reaction. The influence of ten different fixatives on the content of thyroglobulin immobilized on nitrocellulose filters and in single cells and the influence of thyrotropin and interleukin-1β (IL-1β) on the amount of intracellular thyroglobulin were evaluated. The most suitable fixatives for single cells were 2% carbodiimide, Lison's ‘Gendre fluid’ and 2 or 4% paraformaldehyde, whereas Bouin, Carnoy A and B, formalin-calcium and Lillie's formaldehyde-acetic acid-alcohol fixative all resulted in reduction of intracellular thyroglobulin. Two per cent glutaraldehyde caused a considerable reduction (p<0.0001). Nitrocellulose filters were not suitable for evaluation of the fixatives, since the results did not correspond to those obtained with single cells. Thyrotropin (1 U/I) increased intracellular thyroglobulin, whereas addition of interleukin-1β to the culture medium for three days caused a dose-dependent reduction with a plateau level at 2×10−6 gl−1 (104 U/I_ of interleukin-1β. It is concluded that changes in intracellular thyroglobulin concentration caused by either thyrotropin or IL-1β can be quantified under experimental circumstances where samples for measurements of thyroglobulin-mRNA or extracellular thyroglobulin are difficult or impossible to obtain.

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References

  • Conovor, W. J. (1980)Practical nonparametric statistics. 2nd edn. New York: Wiley, & Sons Inc.

    Google Scholar 

  • Dong, Q., Ludgate, M. &Vassart, G. (1989) Towards an antigenic map of human thyroglobulin: identification of ten epitope-bearing sequences within the primary structure of thyroglobulin.J. Endocrinol. 122, 169–76.

    PubMed  Google Scholar 

  • Nagayama, Y., Yamashita, S., Hirayu, H., Izumi, M. Uga, T., Ishikawa, N., Ito, K. &Nagataki, S. (1989) Regulation of thyroid peroxidase and thyroglobulin gene expression by thyrotropin in cultured human thyroid cells.J. Clin. Endocrinol Metab. 68, 1155–9.

    PubMed  Google Scholar 

  • Nibbering, P. H., Leijh, P. C. J. &Van Furth, R. (1985) A cytophotometric method to quantitate the binding of monoclonal antibodies to individual cells.J. Histochem. Cytochem. 33, 453–9.

    PubMed  Google Scholar 

  • Nibbering, P. H., Marijnen, J. G. J., Raap, A. K., Leijh, P. C. J. &Van Furth, R. (1986). Quantitative study of enzyme immunocytochemical reactions performed with enzyme conjugates immobilized on nitrocellulose.Histochemistry 84, 538–43.

    PubMed  Google Scholar 

  • Nibbering, P. H. &Van Furth, R. (1987). Microphotometric quantitation of the reaction product of several indirect immunoperoxidase methods demonstrating monoclonal antibody binding to antigens immobilized on nitrocellulose.J. Histochem. Cytochem 35, 1425–31.

    PubMed  Google Scholar 

  • Paiement, J. &Roy, L. (1988) Electrophoretic protein blots as aids in choosing fixatives for immunocytochemistry.J. Histochem. Cytochem. 36, 441–6.

    PubMed  Google Scholar 

  • Pearse, A. G. E. (1980)Histochemistry: Theoretical and Applied, Vol. 1, 4th edn., pp. 97–158. Edinburgh, London, New York: Churchill Livingstone.

    Google Scholar 

  • Rasmussen, Å. K., Kayser, L., Bech, K., Feldt-Rasmussen, U., Perrild, H. &Bendtzen, K. (1990) Differential effects of interleukin 1α and 1β on cultured human and rat thyroid epithelial cells.Acta Endocrinol. (Copenh).122, 520–6.

    Google Scholar 

  • Svenson, M. &Bendtzen, K. (1988) Inhibitor of interleukin 1 in normal human urine.Scand. J. Immunol. 27, 593–9.

    PubMed  Google Scholar 

  • Yamashita, S., Kimura, H., Ashizawa, K., Nagayama, Y., Hirayu, H., Izumi, M. &Nagataki, S. (1989) Interleukin-1 inhibits thyrotropin induced human thyroglobulin gene expression.J. Endocrinol 122, 177–83.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Medical Anatomy A, The Panum Institute, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen N, Denmark

    Lars Kayser, Birgitte Engelbrecht Olsen & Poul Erik Høyer

  2. Department of Medicine E, Frederiksberg Hospital, Frederiksberg, Denmark

    Lars Kayser

Authors
  1. Lars Kayser
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  2. Birgitte Engelbrecht Olsen
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  3. Poul Erik Høyer
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Kayser, L., Olsen, B.E. & Høyer, P.E. Quantitative cytochemical demonstration of intracellular thyroglobulin in cultured human thyrocytes. Effects of fixatives, TSH and Interleukin-1β. Histochem J 23, 235–240 (1991). https://doi.org/10.1007/BF01462246

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  • DOI: https://doi.org/10.1007/BF01462246

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