Histochemical Approach for Simultaneous Detection of Ectonucleotidase and Alkaline Phosphatase Activities in Tissues

  • Karolina Losenkova
  • Marius Paul
  • Heikki Irjala
  • Sirpa Jalkanen
  • Gennady G. YegutkinEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)


Studies on pathophysiology and the therapeutic potential of extracellular ATP and other purines represent an important and rapidly evolving field. The integral response of the cell is determined by multiple factors, including the release of endogenous ATP, co-expression of different types of nucleotide- and adenosine-selective receptors, as well as the specific makeup of ectoenzymes governing the duration and magnitude of purinergic signaling. Current findings support the presence of an extensive network of purine-converting ectoenzymes that are co-expressed to a variable extent among the mammalian tissues and share similarities in substrate specificity. Here, we describe a histochemical approach for simultaneous detection of ecto-nucleotidase and tissue-nonspecific alkaline phosphatase (TNAP) activities in the same tissue slice. Further employment of this technique for staining human palatine tonsil cryosections revealed selective distribution of the key ectoenzymes within certain tonsillar structures, including germinal centers and connective tissues (ecto-5′-nucleotidase/CD73), as well as interfollicular area (TNAP and NTPDase1/CD39).

Key words

Enzyme histochemistry NTPDase1/CD39 Ecto-5′-nucleotidase/CD73 Tissue-nonspecific alkaline phosphatase Human tonsils 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Karolina Losenkova
    • 1
  • Marius Paul
    • 1
  • Heikki Irjala
    • 2
  • Sirpa Jalkanen
    • 1
  • Gennady G. Yegutkin
    • 1
    Email author
  1. 1.MediCity Research LaboratoryUniversity of TurkuTurkuFinland
  2. 2.Department of Otorhinolaryngology, Head and Neck SurgeryTurku University Hospital and Turku UniversityTurkuFinland

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