Skip to main content
SpringerLink
Log in

Quantification of the cellular proliferation on freshly dispersed cells from rat anterior pituitaries afterin vivo andin vitro labelling with bromodeoxyuridine

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

The labelling index i.e., the proportion of cells in S phase of the cell cycle, has been calculated in cytospin preparations of rat anterior pituitary cells after labelling eitherin vivo orin vitro with the thymidine analogue bromodeoxyuridine (BrdU). The aims of this work were (1) to check whether enzymatic digestion interferes with the incorporation of BrdU into S phase cells and/or whether it has any deletereous effect on the immunohistochemical detection of cells that have already incorporated BrdU, and (2) to check the viability of simultaneous staining for BrdU and markers for the different types of pituitary cells in the cytospins. No statistical difference was found between the labelling index afterin vivo orin vitro labelling with BrdU. Identification of doubly-immunostained cells was straightforward and up to 40% of BrdU-labelled cells were immunopositive for pituitary hormones. It is suggested that cytospin preparations from biopsy samples may be used to study cellular proliferation without exposing the patient to the hazardous effects of BrdU infusion and without the interference of cell culture methods.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Böswald, M., Harasim, S. &Maurer-Schultze, B. (1990). Tracer dose and availability time of thymidine and bromodeoxyuridine: application of bromodeoxyuridine in cell kinetic studies.Cell Tissue Kinet. 23, 169–181.

    PubMed  Google Scholar 

  • Carbajo-Perez, E. &Watanabe, Y. G. (1990). Cellular proliferation in the anterior pituitary of the rat during the postnatal period.Cell Tissue Res. 261, 333–8.

    PubMed  Google Scholar 

  • Carbajo-Perez, E., Motegi, M. &Watanabe, Y. G. (1989). Cell proliferation in the anterior pituitary of mice during growth.Biomed. Res. 10, 275–81.

    Google Scholar 

  • Cocchia, D. &Miani, N. (1980). Immunocytochemical localization of the brain-specific protein S-100 in the pituitary gland of adult rat.J. Neurocytol. 9, 771–82.

    PubMed  Google Scholar 

  • Coltrera, M. D. &Gown, A. M. (1990) PCNA/cyclin expression and BrdU uptake define different subpopulations in different cell lines.J. Histochem. Cytochem. 39, 23–30.

    Google Scholar 

  • Dada, M. O., Campbell, G. T. &Blake, C. A. (1984). Pars distalis cell quantification in normal adult male and female rats.J. Endocrinol. 101, 87–94.

    PubMed  Google Scholar 

  • Danova, M., Riccardi, A., Giordano, M., Girino, M., Mazzini, G., Dezza, I. &Ascari, E. (1988) Cell cycle-related proteins: a flow cytometric study in human tumors.Biol. Cell. 64, 23–8.

    PubMed  Google Scholar 

  • Dawson, A. E., Norton, A. J. &Weinberg, D. B. (1990). Comparative assessment of proliferation and DNA content in breast carcinoma by image analysis and flow cytometry.Am. J. Pathol. 136, 1115–24.

    PubMed  Google Scholar 

  • DeFazio, A., Leary, J. A., Hedley, D. W. &Tattersall, M. H. N. (1987) Immunohistochemical detection of proliferating cells in vivo.J. Histochem. Cytochem. 35, 571–7.

    PubMed  Google Scholar 

  • French, M., Magaud, J. P., Duhaut, P., Bryon, P. A. &Viala, J. J. (1990) Determination of plasma cell labelling index with bromodeoxyuridine using a double immunoenzymatic technique.Stain Tech. 65, 31–6.

    Google Scholar 

  • Garcia, R. L., Coltrera, M. D. &Gown, A. M. (1989). Analysis of proliferative grade using anti-PCNA/cycliun monoclonal antibodies in fixed, embedded tissues. Comparison with flow cytometric analysis.Am. J. Pathol. 134, 733–9.

    PubMed  Google Scholar 

  • Goz, B. (1977) The effect of incorporation of 5-halogenated deoxyuridine into DNA of eukaryotic cells.Pharmacol. Rev. 29, 249–72.

    PubMed  Google Scholar 

  • Gratzner, H. G. (1982). Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: a new reagent for detection for DNA replication.Science 218, 474–5.

    PubMed  Google Scholar 

  • Hedley, D. W. (1989). Flow cytometry using paraffin-embedded tissue: five years on.Cytometry 10, 229–41.

    PubMed  Google Scholar 

  • Hunt, T. E. (1951). The effect of hypophyseal extract on mitotic activity of the rat hypophysis.Anat. Rec. 111, 713–25.

    PubMed  Google Scholar 

  • Hunt, T. E. &Hunt, E. A. (1966). A radioautographic study of the proliferative activity of adrenocortical and hypophyseal cells of the rat at different periods of the estrous cycle.Anat. Rec. 156, 361–8

    Google Scholar 

  • Hymer, W. C., Evans, W. H., Kraicer, J., Mastro, A., Bavis, J. &Griswold, E. (1973). Enrichment of cell types from the rat adenohypophysis by sedimentation at unit gravity.Endocrinology 92, 275–86.

    PubMed  Google Scholar 

  • Inoue, K. &Kurosumi, K. (1981). Mode of proliferation of gonadotropic cells of the anterior pituitary after castration—immunocytochemical and autoradiographic studies.Arch. Histol. Jpn. 44, 71–85.

    PubMed  Google Scholar 

  • Kalbermann, L. E., Szijan, I., Jahn, G. A., Krawiec, L. &Burdman, J. A. (1979). DNA synthesis in the pituitary gland of the rat. Effect of sulpiride and postnatal maturation.Neuroendocrinology 29, 42–8.

    PubMed  Google Scholar 

  • Lloveras, B., Edgerton, S. &Thor, A. D. (1991). Evaluation ofin vitro bromodeoxyuridine labeling of breast carcinomas with the use of a commercial kit.Am. J. Clin. Pathol. 95, 41–7.

    PubMed  Google Scholar 

  • Mastro, A., Shelton, E. &Hymer, W. C. (1969). DNA synthesis in the rat anterior pituitary.J. Cell Biol. 43, 626–9.

    PubMed  Google Scholar 

  • Mcnicol, A. M. &Duffy, A. E. (1987) A study of cell migration in the adrenal cortex of the rat using bromodeoxyuridineCell Tissue Kinet. 20, 519–26.

    PubMed  Google Scholar 

  • McNicol, A. M., Shepherd, M. &Lane, D. P. (1991). Cell proliferation in pituitary adenomas: correlation with hormonal immunoreactivity.J. Endocrinol. Invest. 14 (suppl. 1) P14.

    Google Scholar 

  • McNicol, A. M., Kubba, M. A. G. &McTeague, E. (1988) The mitogenic effects of corticotrophin-releasing factor on the anterior pituitary gland of the rat.J. Endocrinol. 118, 237–41.

    PubMed  Google Scholar 

  • McNicol, A. M., Murray, J. E. &McMeekin, W. (1990) Vasopressin stimulation of cell proliferation in the rat pituitary gland in vitro.J. Endocrinol. 126, 255–9.

    PubMed  Google Scholar 

  • Messier, B. (1969). Effect of exogenous thyroxine on3H-thymidine uptake in mouse pituitary gland.Acta Endocrinol. 61, 133–6.

    PubMed  Google Scholar 

  • Morstyn, G., Kinsella, T., Hsu, S. M., Russo, A., Gratzner, A. &Mitchell, J. (1984). Identification of bromodeoxyuridine in malignant and normal cells following therapy: relationship to complications.Int. J. Radiat. Oncol. Biol. Phys. 10, 1441–6.

    PubMed  Google Scholar 

  • Morstyn, G., Pyke, K., Gardner, J., Ashcroft, R., DeFazio, A. &Bhathal, P. (1986). Immunohistochemical identification of proliferating cells in organ culture using bromodeoxyuridine and a monoclonal antibody.J. Histochem. Cytochem. 34, 697–701.

    PubMed  Google Scholar 

  • Nemoto, R., Uchida, K., Hattori, K., Shimazui, T., Nishijima, Y., Saito, S., Koiso, K. &Harada, M. (1988). S phase fraction of human bladder tumor measured in situ with bromodeoxyuridine labeling.J. Urol. 139, 286–9.

    PubMed  Google Scholar 

  • Nemoto, R. Tohi, Koiso, K. &Harada, M. (1990). S-phase fraction of human prostate adenocarcinomas within vivo bromodeoxyuridine labeling.Cancer 66, 509–14.

    PubMed  Google Scholar 

  • Pan, S., Okazaki, Y. &Takemoto, T. (1990)In vivo cell kinetics studies of the duodenum during and after healing of cysteamine-induced duodenal ulcer in rats using bromodeoxyuridine incorporation.J. Gastroenterol. Hepatol. 5, 281–7.

    PubMed  Google Scholar 

  • Pomerat, G. R. (1941). Mitotic activity in the pituitary of the white rat following castration.Am. J. Anat. 69, 89–121.

    Google Scholar 

  • Ramsdell, J. S. (1990). Thyrotropin-releasing hormone tuoliferation by blocking entry into S phase.Endocrinology 126, 472–9.

    PubMed  Google Scholar 

  • Rosental, D. G., Romano, M. I., Machiavelli, G. A., Grattone, M. L., Perez, R. L., Sterin-Speziale, N. &Burdman, J. A. (1989) Stimulation of cell proliferation in the anterior pituitary gland of the rat by indomethacin.Neuroendocrinol. Lett. 11, 333–8.

    Google Scholar 

  • Silvestrini, R., Costa, A., Veneroni, S., Del Bingo, G. &Persici, P. (1988) Comparative analysis of different approaches to investigate cell kinetics.Cell Tissue Kinet. 21, 123–31.

    PubMed  Google Scholar 

  • Stepién, H. (1981) LHRH inhibits (3H)-thymidine incorporation by pituitary cells culturedin vitro.Endocrinology 109, 1784–6.

    PubMed  Google Scholar 

  • Thiry, M. (1988) Immunoelectron microscope localization of bromodeoxyuridine incorporated into DNA of Ehrlich tumor cell nucleoli.Exp. Cell. Res. 179, 204–13.

    PubMed  Google Scholar 

  • Watanabe, Y. G. &Carbajo-Perez (1990) Cell proliferation in pituitary monolayers as revealed by the BrdU labelling method.Biomed. Res. 11, 373–8.

    Google Scholar 

  • Wright, N. A. &Appleton, D. R. (1980) The metaphase arrest technique. A critical review.Cell Tissue Kinet. 13, 643–63.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy and Histology, Faculty of Medicine, University of Salamanca, Campo Charro s/n, 37007, Salamanca, Spain

    S. Carbajo, F. Gonzalez Del Pozo & E. Carbajo-Perez

Authors
  1. S. Carbajo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Gonzalez Del Pozo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E. Carbajo-Perez
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Carbajo, S., Gonzalez Del Pozo, F. & Carbajo-Perez, E. Quantification of the cellular proliferation on freshly dispersed cells from rat anterior pituitaries afterin vivo andin vitro labelling with bromodeoxyuridine. Histochem J 24, 137–143 (1992). https://doi.org/10.1007/BF01047463

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01047463

Keywords

Search

Navigation