Enzyme-Histochemistry Technique for Visualizing the Dipeptidyl-Peptidase IV (DPP-IV) Activity in the Liver Biliary Tree

  • Vittorio BertoneEmail author
  • Eleonora Tarantola
  • Isabel Freitas
Part of the Methods in Molecular Biology book series (MIMB, volume 1560)


Dipeptidyl-peptidase IV is an enzyme involved in a lot of biochemical processes, where it modifies a number of regulatory proteins by removing the terminal peptides by hydrolysis. Here we describe a histochemical method to demonstrate with accuracy and precision its in situ activity on cryostatic section of Wistar rat liver by means of a simultaneous azo-coupling method.

Key words

Dipeptidyl-peptidase IV Enzyme histochemistry In situ enzyme activity Cryostatic samples 


  1. 1.
    Pro B, Dang NH (2004) CD26/dipeptidyl peptidase IV and its role in cancer. Histol Histopathol 19:1345–1351PubMedGoogle Scholar
  2. 2.
    Ohnuma K, Hosono O, Dang NH, Morimoto C (2011) Dipeptidyl peptidase in autoimmune pathophysiology. Adv Clin Chem 53:51–84CrossRefPubMedGoogle Scholar
  3. 3.
    Gorrell MD, Abbott CA, Kähne T et al (2000) Relating structure to function in the beta-propeller domain of dipeptidylpeptidase IV. Point mutations that influence adenosine deaminase binding, antibody binding and enzyme activity. In: Langner J, Ansorge S (eds) Cellular peptidases in immune functions and diseases 2. Kluwer, New York, NY, pp 89–95Google Scholar
  4. 4.
    Ohnuma K, Inoue H, Uchiyama M, Yamochi T, Hosono O, Dang NH, Morimoto C (2006) T-cell activation via CD26 and caveolin-1 in rheumatoid synovium. Mod Rheumatol 6(1):3–13CrossRefGoogle Scholar
  5. 5.
    Tarantola E, Bertone V, Milanesi G et al (2012) DipeptidylpeptidaseIV, a key enzyme for the degradation of incretins and neuropeptides: activity and expression in the liver of lean and obese rats. Eur J Histochem 56:e41CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Mentlein R (1999) Dipeptidyl-peptidase IV (CD26)-role in the inactivation of regulatory peptides. Regul Pept 85:9–24CrossRefPubMedGoogle Scholar
  7. 7.
    Mentlein R (2009) Mechanisms underlying the rapid degradation and elimination of the incretin hormones GLP-1 and GIP. Best Pract Res Clin Endocrinol Metab 4:443–452CrossRefGoogle Scholar
  8. 8.
    Hildebrandt M, Rose M, Mayr C et al (2000) Dipeptidyl peptidase IV (DPP IV, CD26) in patients with mental eating disorders. Adv Exp Med Biol 477:197–204CrossRefPubMedGoogle Scholar
  9. 9.
    Gorrell MD (2005) Dipeptidyl peptidase IV and related enzymes in cell biology and liver disorders. Clin Sci 108:1–16CrossRefGoogle Scholar
  10. 10.
    Itou M, Kawaguchi T, Taniguchi E, Sata M (2013) Dipeptidyl peptidase-4: a key player in chronic liver disease. World J Gastroenterol 19:2298–2306CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Miyazaki M, Kato M, Tanaka K et al (2012) Increased hepatic expression of dipeptidyl peptidase-4 in non-alcoholic fatty liver disease and its association with insulin resistance and glucose metabolism. Mol Med Report 5:729–733Google Scholar
  12. 12.
    Williams KH, Shackel NA, Gorrell MD et al (2013) Diabetes and nonalcoholic fatty liver disease: a pathogenic duo. Endocr Rev 34:84–129CrossRefPubMedGoogle Scholar
  13. 13.
    Kesty NC, Roth JD, Maggs D (2008) Hormone based therapies in the regulation of fuel metabolism and body weight. Expert Opin Biol Ther 8:1733–1747CrossRefPubMedGoogle Scholar
  14. 14.
    Chyan YJ, Chuang LM (2007) Dipeptidyl peptidase-IV inhibitors: an evolving treatment for type 2 diabetes from the incretin concept. Recent Pat Endocr Metab Immun Drug Discov 1:15–24CrossRefGoogle Scholar
  15. 15.
    DiStefano JK, Watanabe RM (2010) Pharmacogenetics of anti-diabetes drugs. Pharmaceuticals (Basel) 3:2610–2646CrossRefGoogle Scholar
  16. 16.
    Martin JH, Deacon CF, Gorrell MD, Prins JB (2011) Incretin-based therapies-review of the physiology, pharmacology and emerging clinical experience. Intern Med J 41:299–307CrossRefPubMedGoogle Scholar
  17. 17.
    Bergmann K, Sypniewska G (2013) Diabetes as a complication of adipose tissue dysfunction. Is there a role for potential new biomarkers? Clin Chem Lab Med 51:177–185CrossRefPubMedGoogle Scholar
  18. 18.
    Piazza GA, Callanan HM, Mowery J et al (1989) Evidence for a role of dipeptidyl peptidase IV in fibronectin-mediated interactions of hepatocytes with extracellular matrix. Biochem J 262:327–334CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Balaban YH, Korkusuz P, Simsek H et al (2007) Dipeptidyl peptidase IV (DDP IV) in NASH patients. Ann Hepatol 6(4):242–250PubMedGoogle Scholar
  20. 20.
    Wang XM, Yao TW, Nadvi NA et al (2008) Fibroblast activation protein and chronic liver disease. Front Biosci 13:3168–3180CrossRefPubMedGoogle Scholar
  21. 21.
    Gorrell MD, Gysbers V, McCaughan GW (2001) CD26: a multifunctional integral membrane and secreted protein of activated lymphocytes. Scand J Immunol 54:249–264CrossRefPubMedGoogle Scholar
  22. 22.
    Gonzalez-Gronow M, Hershfield MS, Arredondo-Vega FX et al (2001) Cell surface adenosine deaminase binds and stimulates plasminogen activation on 1-LN human prostate cancer cells. J Biol Chem 279:20993–20998CrossRefGoogle Scholar
  23. 23.
    Matheeussen V, Jungraithmayr W, De Meester I (2012) Dipeptidyl peptidase 4 as a therapeutic target in ischemia/reperfusion injury. Pharmacol Ther 136:267–282CrossRefPubMedGoogle Scholar
  24. 24.
    Ou X, O’Leary HA, Broxmeyer HE (2013) Implications of DPP4 modification of proteins that regulate stem/progenitor and more mature cell types. Blood 122:161–169CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Batra S, Kuo C, Rakusan K (1989) Spatial distribution of coronary capillaries: A-V segment staggering. Adv Exp Med Biol 248:241–247CrossRefPubMedGoogle Scholar
  26. 26.
    Batra S, Konig MF, Cruz-Orives LM (1995) Unbiased estimation of capillary length from vertical slices. J Microscopy 178(Pt 2):152–159CrossRefGoogle Scholar
  27. 27.
    Dimitrova D, Tashkova S, Dikov A et al (2002) Enzyme activities of human milk cells. Comptes rendus de l’Academie Bulgare des Sciences, Tome, p 55Google Scholar
  28. 28.
    Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry.A laboratory manual. Springer, BerlinCrossRefGoogle Scholar
  29. 29.
    Van Noorden CJF, Fredericks WM (1992) Enzyme histochemistry. A laboratory manual of current methods. Oxford Science Publications. Oxford University Press – Royal Microscopy Society, OxfordGoogle Scholar
  30. 30.
    Horobin RW (1982) Histochemistry. Pag 135, Fig. 47. Fisher & Butterworths edsGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Vittorio Bertone
    • 1
    Email author
  • Eleonora Tarantola
    • 1
  • Isabel Freitas
    • 1
  1. 1.Department of Biology and Biotechnology “Lazzaro Spallanzani”University of PaviaPaviaItaly

Personalised recommendations