Abstract
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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Ralevic V, Burnstock G (1998) Receptors for purines and pyrimidines. Pharmacol Rev 50(3):413–492
Yegutkin GG (2008) Nucleotide- and nucleoside-converting ectoenzymes: important modulators of purinergic signalling cascade. Biochim Biophys Acta 1783(5):673–694
Yegutkin GG (2014) Enzymes involved in metabolism of extracellular nucleotides and nucleosides: Functional implications and measurement of activities. Crit Rev Biochem Mol Biol 49(6):473–497
Zimmermann H, Zebisch M, Strater N (2012) Cellular function and molecular structure of ecto-nucleotidases. Purinergic Signal 8(3):437–502
Eltzschig HK, Sitkovsky MV, Robson SC (2012) Purinergic signaling during inflammation. N Engl J Med 367(24):2322–2333
Kauffenstein G, Drouin A, Thorin-Trescases N, Bachelard H, Robaye B, D’Orleans-Juste P, Marceau F, Thorin E, Sevigny J (2010) NTPDase1 (CD39) controls nucleotide-dependent vasoconstriction in mouse. Cardiovasc Res 85(1):204–213
Buchet R, Millan JL, Magne D (2013) Multisystemic functions of alkaline phosphatases. Methods Mol Biol 1053:27–51
Yegutkin GG, Auvinen K, Rantakari P, Hollmen M, Karikoski M, Grenman R, Elima K, Jalkanen S, Salmi M (2015) Ecto-5′-nucleotidase/CD73 enhances endothelial barrier function and sprouting in blood but not lymphatic vasculature. Eur J Immunol 45(2):562–573
Langer D, Hammer K, Koszalka P, Schrader J, Robson S, Zimmermann H (2008) Distribution of ectonucleotidases in the rodent brain revisited. Cell Tissue Res 334(2):199–217
Maj T, Wang W, Crespo J, Zhang H, Wei S, Zhao L, Vatan L, Shao I, Szeliga W, Lyssiotis C, Liu JR, Kryczek I, Zou W (2017) Oxidative stress controls regulatory T cell apoptosis and suppressor activity and PD-L1-blockade resistance in tumor. Nat Immunol 18(12):1332–1341
Allard B, Longhi MS, Robson SC, Stagg J (2017) The ectonucleotidases CD39 and CD73: novel checkpoint inhibitor targets. Immunol Rev 276(1):121–144
Wachstein M, Meisel E (1957) Histochemistry of hepatic phosphatases of a physiologic pH; with special reference to the demonstration of bile canaliculi. Am J Clin Pathol 27(1):13–23
Street SE, Kramer NJ, Walsh PL, Taylor-Blake B, Yadav MC, King IF, Vihko P, Wightman RM, Millan JL, Zylka MJ (2013) Tissue-nonspecific alkaline phosphatase acts redundantly with PAP and NT5E to generate adenosine in the dorsal spinal cord. J Neurosci 33(27):11314–11322
Mercier N, Kiviniemi TO, Saraste A, Miiluniemi M, Silvola J, Jalkanen S, Yegutkin GG (2012) Impaired ATP-induced coronary blood flow and diminished aortic NTPDase activity precede lesion formation in apolipoprotein E-deficient mice. Am J Pathol 180(1):419–428
Yegutkin GG, Guerrero-Toro C, Kilinc E, Koroleva K, Ishchenko Y, Abushik P, Giniatullina R, Fayuk D, Giniatullin R (2016) Nucleotide homeostasis and purinergic nociceptive signaling in rat meninges in migraine-like conditions. Purinergic Signal 12(3):561–574
Aliagas E, Vidal A, Torrejon-Escribano B, Taco Mdel R, Ponce J, de Aranda IG, Sevigny J, Condom E, Martin-Satue M (2013) Ecto-nucleotidases distribution in human cyclic and postmenopausic endometrium. Purinergic Signal 9(2):227–237
Villamonte ML, Torrejon-Escribano B, Rodriguez-Martinez A, Trapero C, Vidal A, Gomez de Aranda I, Sevigny J, Matias-Guiu X, Martin-Satue M (2018) Characterization of ecto-nucleotidases in human oviducts with an improved approach simultaneously identifying protein expression and in situ enzyme activity. Histochem Cell Biol 149(3):269–276
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Losenkova, K., Paul, M., Irjala, H., Jalkanen, S., Yegutkin, G.G. (2020). Histochemical Approach for Simultaneous Detection of Ectonucleotidase and Alkaline Phosphatase Activities in Tissues. In: Pelegrín, P. (eds) Purinergic Signaling. Methods in Molecular Biology, vol 2041. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9717-6_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9717-6_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9716-9
Online ISBN: 978-1-4939-9717-6
eBook Packages: Springer Protocols