title: Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes In Vitro.
abstract: Extracellular ATP (eATP) acts as a danger-associated molecular pattern which induces reactive response of astrocytes after brain insult, including morphological remodeling of astrocytes, proliferation, chemotaxis, and release of proinflammatory cytokines. The responses induced by eATP are under control of ecto-nucleotidases, which catalyze sequential hydrolysis of ATP to adenosine. In the mammalian brain, ecto-nucleotidases comprise three enzyme families: ecto-nucleoside triphosphate diphosphohydrolases 1-3 (NTPDase1-3), ecto-nucleotide pyrophosphatase/phospodiesterases 1-3 (NPP1-3), and ecto-5'-nucleotidase (eN), which crucially determine ATP/adenosine ratio in the pericellular milieu. Altered expression of ecto-nucleotidases has been demonstrated in several experimental models of human brain dysfunctions. In the present study, we have explored the pattern of NTPDase1-3, NPP1-3, and eN expression by cultured cortical astrocytes challenged with 1 mmol/L ATP (eATP). At the transcriptional level, eATP upregulated expression of NTPDase1, NTPDase2, NPP2, and eN, while, at translational and functional levels, these were paralleled only by the induction of NTPDase2 and eN. Additionally, eATP altered membrane topology of eN, from clusters localized in membrane domains to continuous distribution along the cell membrane. Our results suggest that eATP, by upregulating NTPDase2 and eN and altering the enzyme membrane topology, affects local kinetics of ATP metabolism and signal transduction that may have important roles in the process related to inflammation and reactive gliosis.
authors.affiliation: Institute for Physiology and Biochemistry, Faculty of Biology, University of Belgrade, Studentski trg 3, Belgrade, 11001, Serbia.
authors.forename: Dusica
authors.initials: D
authors.lastname: Brisevac
authors.affiliation: Institute for Physiology and Biochemistry, Faculty of Biology, University of Belgrade, Studentski trg 3, Belgrade, 11001, Serbia.|Center for Laser Microscopy, Faculty of Biology, University of Belgrade, Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Adzic
authors.affiliation: Institute for Physiology and Biochemistry, Faculty of Biology, University of Belgrade, Studentski trg 3, Belgrade, 11001, Serbia.
authors.forename: Danijela
authors.initials: D
authors.lastname: Laketa
authors.affiliation: Institute for Biological Research "Sinisa Stankovic", University of Belgrade, Belgrade, Serbia.
authors.forename: Ana
authors.initials: A
authors.lastname: Parabucki
authors.affiliation: Institute for Physiology and Biochemistry, Faculty of Biology, University of Belgrade, Studentski trg 3, Belgrade, 11001, Serbia.|Center for Laser Microscopy, Faculty of Biology, University of Belgrade, Belgrade, Serbia.
authors.forename: Milena
authors.initials: M
authors.lastname: Milosevic
authors.affiliation: Institute for Biological Research "Sinisa Stankovic", University of Belgrade, Belgrade, Serbia.
authors.forename: Irena
authors.initials: I
authors.lastname: Lavrnja
authors.affiliation: Institute for Biological Research "Sinisa Stankovic", University of Belgrade, Belgrade, Serbia.
authors.forename: Ivana
authors.initials: I
authors.lastname: Bjelobaba
authors.affiliation: Département de microbiologie-infectiologie et d'immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, Canada.|Centre de recherche du CHU de Québec Université Laval, Québec, QC, G1V 4G2, Canada.
authors.forename: Jean
authors.initials: J
authors.lastname: Sévigny
authors.affiliation: Institute of Neuroanatomy, Faculty of Medicine, RWTH Aachen University, Aachen, Germany.|Department of Anatomy II, Ludwig-Maximilians-University of Munich, Munich, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Kipp
authors.affiliation: Institute for Physiology and Biochemistry, Faculty of Biology, University of Belgrade, Studentski trg 3, Belgrade, 11001, Serbia. nnedel@bio.bg.ac.rs.
authors.forename: Nadezda
authors.initials: N
authors.lastname: Nedeljkovic
mesh_terms: D015720:5'-Nucleotidase / Q000096:biosynthesis* / Q000235:genetics; D000241:Adenosine / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D002455:Cell Division; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004790:Enzyme Induction / Q000187:drug effects; D005911:Gliosis / Q000201:enzymology; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D010727:Phosphoric Diester Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D061251:Primary Cell Culture; D014176:Protein Biosynthesis / Q000187:drug effects; D011755:Pyrophosphatases / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D000099189:Lysophospholipase D
chemical_list: D015720:5'-Nucleotidase; D000241:Adenosine; D000255:Adenosine Triphosphate; D009419:Nerve Tissue Proteins; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D000099189:Lysophospholipase D; C524442:Enpp3 protein, rat; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1
keywords: Ecto-5′-nucleotidase/CD73; Ecto-nucleotidase; Extracellular ATP; NTPDase2/CD39L1; Reactive astrogliosis