Skip to main content
Log in

Carbonic anhydrase I and II as a differentiation marker of human and rat colonic enterocytes

  • Published:
Research in Experimental Medicine

Abstract

Carbonic anhydrase (CA) is an enzyme that is expressed in the intestine and catalyzes the reversible hydration of CO2 in the following reaction: CO2+H2O⇄H2CO3⇄H++HCO 3 . To elucidate the association of CA expression with the differentiation of colonic enterocytes, we investigated the expression and localization of CA using a Northern blotting analysis, Western blotting analysis, and immunohistochemical staining. A Northern blotting analysis revealed an abundant expression of CA I and II mRNA in the colonic epithelial cells. However, the expression of CA III mRNA was not detected. According to the results of immunohistochemical staining of the human colonic mucosa using antisera against CA I and II, both CA I and II were localized on the cytoplasm of non-goblet columnar cells in the upper half of the crypts where more differentiated cells are located. According to the results of immunohistochemical staining of the rat colonic mucosa, neither CA I and II were detected at the new-born stage. The expression of CAs in the upper half of the crypts began to rise from 1 week after birth, and thereafter increased according to the growth of the rats. At 3 weeks after birth, the expression of CAs was almost the same as that of the adult rats. The amount of CA proteins evaluated by a Western blotting analysis revealed that the expression of CAs increased gradually until reaching a maximum level at 6 or 8 weeks. These results therefore suggest that CA I and II appear to be good markers for the differentiation of enterocytes in the colonic mucosa.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69: 1408–1411

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Belzen NV, Diesveld MPG, Van Der Made ACJ, Nozawa Y, Dinjens WNM, Vlietstra R, Trapman J, Bosman FT (1995) Identification of mRNAs that show modulated expression during colon carcinoma cell differentiation. Eur J Biochem 234:843–848

    Article  PubMed  Google Scholar 

  3. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299

    Article  CAS  PubMed  Google Scholar 

  4. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    Article  CAS  PubMed  Google Scholar 

  5. Fleming RE, Crouch EC, Ruzicka CA, Sly WS (1993) Pulmonary carbonic anhydrase IV: developmental regulation and cell-specific expression in the capillary endothelium. Am J Physiol 265:L627–L635

    CAS  PubMed  Google Scholar 

  6. Gramlich TL, Hennigar RA, Spicer SS, Schulte BA (1990) Immunohistochemical localization of sodium-potassium-stimulated adenosine triphosphatase and carbonic anhydrase in human colon colonic neoplasms. Arch Pathol Lab Med 114:415–419

    CAS  PubMed  Google Scholar 

  7. Gudmar L, Selking O, Wistrand PJ (1985) Amount and distribution of carbonic anhydrase CA I and CA II in the gastrointestinal tract. Gastroenterology 88:1151–1161

    Google Scholar 

  8. Lacy ER, Colony PC (1985) Localization of carbonic anhydrase activity in the developing rat colon. Gastroenterology 89:138–150

    CAS  PubMed  Google Scholar 

  9. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227:680–685

    Article  CAS  Google Scholar 

  10. Lloyd J, Brownson C, Tweedie S, Charlton J, Edwards YH (1987) Human muscle carbonic anhydrase: gene structure and DNA methylation patterns in fetal and adult tissues. Genes Dev 1:594–602

    Article  CAS  PubMed  Google Scholar 

  11. Mack DR, Hollingsworth MA (1994) Alteration in expression of MUC2 and MUC3 mRNA levels in HT29 colonic carcinoma cells. Biochem Biophys Res Commun 199:1012–1018

    Article  CAS  PubMed  Google Scholar 

  12. Mori M, Staniunas RJ, Barnard GF, Jessup JM, Steele GD, Chen LB (1993) The significance of carbonic anhydrase expression in human colorectal cancer. Gastroenterology 105:820–826

    CAS  PubMed  Google Scholar 

  13. Mavilio F, Giampaolo A, Care A, Migliaccio G, Calandrini M, Russo G, Pagliardi GL, Mastroberardino G, Marinucci M, Perchle C (1983) Molecular mechanisms of human hemoglobin switching: selective undermethylation and expression of globin genes in embryonic, fetal, and adult erythroblasts. Proc Natl Acad Sci USA 80:6907–6911

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Potten C S, Loeffler M (1990) Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the crypt. Development 110:1001–1020

    CAS  PubMed  Google Scholar 

  15. Potten C S, Schofield R, Lajtha LG (1979) A comparison of cell replacement in bone marrow, testis and three regions of surface epithelium. Biochim Biophys Acta 560:281–299

    CAS  PubMed  Google Scholar 

  16. Sly WS, Hu PY(1995) Human carbonic anhydrases and carbonic anhydrase deficiencies. Annu Rev Biochem 64:375–401

    Article  CAS  PubMed  Google Scholar 

  17. Sowden J, Leigh S, Talbot I, Delhanty J, Edwards Y (1993) Expression from the proximal promoter of the carbonic anhydrase I gene as a marker for differentiation in colon epithelia. Differentiation 53:67–74

    Article  CAS  PubMed  Google Scholar 

  18. Stamatoyannopoulos G (1991) Human globin gene switching. Science 252:383

    Article  CAS  PubMed  Google Scholar 

  19. Tashian RE, Venta PJ, Nicewanden PH, Hewett-Emmett D (1990) Evolution, structure, and expression of the carbonic anhydrase multigene family. In: Ogita Z, Markert CL (eds) Isozymes: structure, function, and use in biology and medicine. Wiley-Liss, New York, p159–175

    Google Scholar 

  20. Towbin H, Staehlin T, Gordon J (1979) Electrophoretic transfer of protein polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sci USA 76:4350–4354

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Velcich V, Palumbo L, Jarry A, Laboisse C, Racevslis J, Augenlicht L (1995) Patterns of expression of lineage-specific markers during the in vitro-induced differentiation of HT29 colon carcinoma cells. Cell Growth Differentiation 6:749–757

    CAS  PubMed  Google Scholar 

  22. Villeval JL, Testa U, Vinci G, Tonthat H, Bettaieb A, Titeux M, Cramer P, Edelman L, Rochant H, Breton-Gorius J, Vainchenker W (1985) Carbonic anhydrase I is an early specific marker of normal human erythroid differentiation. Blood 66:1162–1170

    CAS  PubMed  Google Scholar 

  23. Yokoyama S, Shatney CH, Mochizuki H, Hase K, Johnson DL, Cummings S, Trollope ML, Tamakuma S (1997) The potential role of fecal carbonic anhydrase II in screening for colorectal cancer. Am Surg 63:243–247

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bekku, S., Mochizuki, H., Takayama, E. et al. Carbonic anhydrase I and II as a differentiation marker of human and rat colonic enterocytes. Res. Exp. Med. 198, 175–185 (1998). https://doi.org/10.1007/s004330050101

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s004330050101

Key words

Navigation