Skip to main content
SpringerLink
Log in

Changes of colon epithelium proliferation due to individual aging with cyclin proliferating cell nuclear antigen (PCNA/cyclin) immunostaining compared to [3H]-thymidine radioautography

  • Original Paper
  • Published:
Histochemistry Aims and scope Submit manuscript

Abstract

We report a change in the proliferative activity of mouse colonic epithelium due to development and aging. In order to measure the proliferative activity, colonic epithelium was immunostained for cyclin proliferating cell nuclear antigen (PCNA/cyclin), which appears from the Gl to the S phase of the cell cycle, and compared with labeling obtained by [3H]-thymidine radioautography. Litter mice of six age groups from the fetal period (embryonic day 19), newborn period (postnatal day 1), suckling period (postnatal day 5), weaning period (postnatal dy 21), adult period (2 month old) to the senescent period (11 month old) were examined by immunohistochemistry. The descending colons were fixed in methacarn (method-Carnoy) and embedded in paraffin. Sections were stained for PCNA/cyclin activity using 19A2 monoclonal antibody and the avidin-biotin peroxidase complex (ABC) technique. For radioautography, litter mice of nine age groups using in vivo intraperitoneal administration of [3H]-thymidine. The labeling indices of colonic epithelial cells in the proliferative zone were then analyzed and compared between the two investigative methods. Our results show that the prliferative activity of mice colon was high in the fetal and newborn periods and almost constant from the suckling period to senescence, as demonstrated by both PCNA/cyclin immunohistochemistry and [3H]-thymidine radioautography. The labeling index seen by PCNA/cyclin immunohistochemistry was, however, higher than that seen by [3H]-thymidine radioautography.

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

  • Batt ER (1969) Sulfate accumulation by mouse intestine: influence of age and other factors. Am J Physiol 217:1101–1104

    Google Scholar 

  • Bravo R (1986) Synthesis of the nuclear protein cyclin (PCNA) and its relationship with DNA replication. Exp Cell Res 163:287–293

    Google Scholar 

  • Bravo R, Frank R, Blundell PA (1987) Cyclin/PCNA is the auxiliary protein DNA polymerase-δ. Nature 326:515–517

    Google Scholar 

  • Cairnie AB, Lamerton LF, Steel GG (1965a) Cell proliferation studies in the intestinal epithelium of the rat. I. Demonstration of the kinetic parameters. Exp Cell Res 39:528–538

    Google Scholar 

  • Cairnie AB, Lamerton LF, Steel GG (1965b) Cell proliferation studies in the intestinal epithelium of the rat. II. Theoretical aspects. Exp Cell Res 39:539–553

    Google Scholar 

  • Celis JE, Celis A (1985) Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Cell Biol 82:3262–3266

    Google Scholar 

  • Chang WWL, Leblond CP (1971a) Renewal of the epithelium in the descending colon of the mouse. I. Presence of three cell populations: vacuolated-columnar, mucous and argentaffin. Am J Anat 131:73–100

    Google Scholar 

  • Chang WWL, Leblond CP (1971b) Renewal of the epithelium in the descending colon of the mouse. II. Renewal of argentaffin cells. Am J Anat 131:101–110

    Google Scholar 

  • Chang WWL, Leblond CP (1971c) Renewal of the epithelium in the descending colon of the mouse. III. Diurnal variation in the proliferative activity of epithelial cells. Am J Anat 131:111–120

    Google Scholar 

  • Chang WWL, Nadler NJ (1975) Renewal of the epithelium in the descending colon of the mouse. IV. Cell population kinetics of vacuolated- columnar and mucous cells. Am J Anat 144:39–56

    Google Scholar 

  • Cheng H, Bjerknes M (1982) Whole population cell kinetics of mouse duodenal, jejunal, ileal, and colonic epithelia as determined by radioautography and flow cytometry. Anat Rec 203:251–264

    Google Scholar 

  • Colony PC, Neutra MR (1983) Epithelial differentiation in the fetal rat colon. I. Plasma membrane phosphatase activities. Dev Biol 97:349–363

    Google Scholar 

  • Coltrera MD, Gown AM (1991) PCNA/cyclin expression and BrdU uptake define different subpopulations in different cell lines. J Histochem Cytochem 39:23–30

    Google Scholar 

  • Danovitch SH, Laster L (1969) The development of arylsulfatase in the small intestine of the rat. Biochem J 114:343–350

    Google Scholar 

  • Diebold J, Lai MD, Löhrs U (1992) Analysis of proliferative activity in colorectal mucosa by immunohistochemical detection of proliferating cell nuclear antigen (PCNA). Virchows Arch B Cell Pathol 62:283–289

    Google Scholar 

  • Dierendonck JH van, Wijsman JH, Keijzer R, Velde CJH van de, Cornelisse CJ (1991) Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdU labelling and Ki-67 staining. Am J Pathol 138:1165–1172

    Google Scholar 

  • Fielding LP, Fenoglio-Preiser CM, Freedman LS (1992) The future of progenostic factors in outcome prediction for patients with cancer. Cancer 70:2367–2377

    Google Scholar 

  • Galand P, Degraef C (1989) Cyclin/PCNA immunostaining as an alternatives to tritiated thymidine pulse labelling for marking S phase cells in paraffin sections from animals and human tissues. Cell Tissue Kinet 22:383–392

    Google Scholar 

  • Galand P, Mainguet P, Arguello M, Chretien J, Douxfils N (1968) In vitro autoradiographic studies of cell proliferation in the gastrointestinal tract of man. J Nuclear Med 9:37–39

    Google Scholar 

  • Garcia RL, Coltrera MD, Gown AM (1989) Analysis of proliferative grade using anti- PCNA/cyclin monoclonal antibodies in fixed, embedded tissues. Comparison with cytometric analysis. Am J Pathol 134:733–739

    Google Scholar 

  • Goldstein R, Klein T, Freier S, Menczel J (1971) Alkaline phosphatase and disaccharidase activities in the rat intestine from birth to weaning. I. Effect of diet on enzyme development. Am J Clin Nutr 24:1224–1231

    Google Scholar 

  • Helander HF (1973) Morphological studies on the development of the rat colonic mucosa. Acta Anat 85:153–176

    Google Scholar 

  • Hsu SM, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580

    Google Scholar 

  • James JT, Shamsuddin AM, Trump BF (1982) A comparative study of the normal histochemical and proliferative properties of the large intestine in ICR/Ha and C57BI/Ha mice. Virchows Arch B Cell Pathol 41:133–144

    Google Scholar 

  • Kamel OW, LeBrun DP, Davis RE, Berry GJ, Warnke RA (1991) Growth fraction estimation of malignant lymphomas in formalin-fixed paraffin-embedded tissue using anti-PCNA/cyclin 19A2. Comparison with Ki-67 labeling. Am J Pathol 138:1471–1477

    Google Scholar 

  • Kawachi T, Tanaka N, Kogure K, Sugimura T (1973) Changes in aldolase isozymes of the digestive tract during postnatal development. Biochim Biophys Acta 320:59–63

    Google Scholar 

  • Kunkel TA, Sabatino RD, Bambara RA (1987) Exonucleolytic proofreading by calf thymus DNA polymerase-δ. Proc Natl Acad Sci USA 84:4865–4869

    Google Scholar 

  • Lacy ER (1991) Functional morphology of the large intestine. In: Shultz SG, Field M, Frizzell RA, Rauner BB (eds) Handbook of physiology, section 6: The gastrointestinal system, vol IV. American Physiological Society, Bethesda, Maryland, pp 152–173

    Google Scholar 

  • Morret-Rauis M, Degraef C, Galand P (1990) S phase duration measurement by combined PCNA/cyclin immunostaining and radioautography after a single pulse-labelling with 3H-thymidine. Cell Biol Int Rep 14:765–774

    Google Scholar 

  • Nagata T (1990) Principles and techniques of electron microscopic radioautography. In: National training course on the application of electron microscopy in biomedical research and diagnosis of human diseases. The World Health Organization, Beijing, pp 139–158

    Google Scholar 

  • Ogata K, Kurki P, Celis JE, Nakamura RM, Tan EM (1987) Monoclonal antibodies to a nuclear protein (PCNA/cyclin) associated DNA replication. Exp Cell Res 168:475–486

    Google Scholar 

  • Pace JL (1971) The age of appearance of the haustra of the human colon. J Anat 109:75–80

    Google Scholar 

  • Prelich G, Kostura M, Marshak DR, Mathews MB, Stillman B (1987) The cell-cycle regulated proliferatingnuclear antigen is required for SV40 DNA replication in vitro. Nature 326:471–475

    Google Scholar 

  • Puchtler H, Waldrop FS, Meloan SN, Terry MS, Conner HM (1970) Methacarn (methanol-Carnoy) fixation. Practical and theoretical considerations. Histochemie 21:97–116

    Google Scholar 

  • Raymond WA, Leong AS-Y (1989) Nucleolar organizer regions relate to growth fractions in human breast carcinoma. Hum Pathol 20:741–746

    Google Scholar 

  • Sakata T, Engelhardt W von (1981) Luminal mucin in the large intestine of mice, rats, and guinea pigs. Cell Tissue Res 219:629–635

    Google Scholar 

  • Sheahan DG, Jervis HR (1976) Comparative histochemistry of gastrointestinal mucosubstances. Am J Anat 146:103–132

    Google Scholar 

  • Shimizu T, Usuda N, Yamanda T, Sugenoya A, Iida F (1993) Proliferative activity of human thyroid tumors evaluated by proliferating cell nuclear antigen/cyclin immunohistochemical studies. Cancer 71:2807–2812

    Google Scholar 

  • Taatjes DJ, Roth J (1990) Selectives loss of sialic acid from rat small intestinal epithelial cells during postnatal development: demonstration with lectin-gold techniques. Eur J Cell Biol 53:255–266

    Google Scholar 

  • Tabuchi K, Honda C, Nakane PK (1987) Demonstration of proliferating cell nuclear antigen (PCNA/cyclin) in glioma cells. Neurol Med Chir (Tokyo) 27:1–5

    Google Scholar 

  • Takahashi H, Oishi Y, Chuang S-S, Morii S (1992) Effects of tissue fixation and processing on proliferationg cell nuclear antigen (PCNA) immunohistochemistry. Acta Pathol Jpn 48:621–623

    Google Scholar 

  • Tsubouchi S (1981) Kinetic analysis of epithelial cell migration in the mouse descending colon. Am J Anat 161:239–246

    Google Scholar 

  • Yu CC-W, Woods AL, Levison DA (1992) The assessment of cellular proliferation by immunohistochemistry: a review of currently available methods and their applications. Histochem J 24:121–131

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Shinshu University School of Medicine, 3-1-1 Asahi, 390, Matsumoto, Japan

    T. Nagata, T. Morita, N. Usuda & T. Hanai

Authors
  1. T. Nagata
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Morita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N. Usuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T. Hanai
    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

Nagata, T., Morita, T., Usuda, N. et al. Changes of colon epithelium proliferation due to individual aging with cyclin proliferating cell nuclear antigen (PCNA/cyclin) immunostaining compared to [3H]-thymidine radioautography. Histochemistry 101, 13–20 (1994). https://doi.org/10.1007/BF00315826

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00315826

Keywords

Search

Navigation