Targeted disruption of cd73/ecto-5′-nucleotidase alters thromboregulation and augments vascular inflammatory response

Patrycja Koszalka, Burcin Özüyaman, Yuqing Huo, Alma Zernecke, Ulrich Flögel, Norbert Braun, Anja Buchheiser, Ulrich K.M. Decking, Michael L. Smith, Jean Sévigny, Adrian Gear, Artur Aron Weber, Andrei Molojavyi, Zhaoping Ding, Christian Weber, Klaus Ley, Herbert Zimmermann, Axel Gödecke, Jürgen Schrader

Research output: Contribution to journalArticle

182 Citations (Scopus)

Abstract

To investigate the role of adenosine formed extracellularly in vascular homeostasis, mice with a targeted deletion of the cd73/ecto-5′- nucleotidase were generated. Southern blot, RT-PCR, and Western blot analysis confirmed the constitutive knockout. In vivo analysis of hemodynamic parameters revealed no significant differences in systolic blood pressure, ejection fraction, or cardiac output between strains. However, basal coronary flow measured in the isolated perfused heart was significantly lower (-14%; P<0.05) in the mutant. Immunohistochemistry revealed strong CD73 expression on the endothelium of conduit vessels in wild-type (WT) mice. Time to carotid artery occlusion after ferric chloride (FeCl3) was significantly reduced by 20% in cd73-/- mice (P<0.05). Bleeding time after tail tip resection tended to be shorter in cd73-/- mice (-35%). In vivo platelet cAMP levels were 0.96±0.46 in WT versus 0.68±0.27 pmol/106 cells in cd73-/- mice (P<0.05). Under in vitro conditions, platelet aggregation in response to ADP (0.05 to 10 μmol/L) was undistinguishable between the two strains. In the cremaster model of ischemia-reperfusion, the increase in leukocyte attachment to endothelium was significantly higher in cd73-/- compared with WT littermates (WT 98% versus cd73-/- 245%; P<0.005). The constitutive adhesion of monocytes in ex vivo-perfused carotid arteries of WT mice was negligible but significantly increased in arteries of cd73-/- mice (P<0.05). Thus, our data provide the first evidence that adenosine, extracellularly formed by CD73, can modulate coronary vascular tone, inhibit platelet activation, and play an important role in leukocyte adhesion to the vascular endothelium in vivo.

Original languageEnglish (US)
Pages (from-to)814-821
Number of pages8
JournalCirculation Research
Volume95
Issue number8
DOIs
StatePublished - Oct 15 2004
Externally publishedYes

Fingerprint

5'-Nucleotidase
Blood Vessels
Carotid Arteries
Adenosine
Endothelium
Leukocytes
Blood Pressure
Bleeding Time
Platelet Activation
Vascular Endothelium
Southern Blotting
Platelet Aggregation
Cardiac Output
Adenosine Diphosphate
Reperfusion
Tail
Monocytes
Homeostasis
Blood Platelets
Ischemia

Keywords

  • Adenosine
  • Ecto-5′-nucleotidase
  • Thrombosis
  • Transgenic mice
  • Vascular inflammation

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Targeted disruption of cd73/ecto-5′-nucleotidase alters thromboregulation and augments vascular inflammatory response. / Koszalka, Patrycja; Özüyaman, Burcin; Huo, Yuqing; Zernecke, Alma; Flögel, Ulrich; Braun, Norbert; Buchheiser, Anja; Decking, Ulrich K.M.; Smith, Michael L.; Sévigny, Jean; Gear, Adrian; Weber, Artur Aron; Molojavyi, Andrei; Ding, Zhaoping; Weber, Christian; Ley, Klaus; Zimmermann, Herbert; Gödecke, Axel; Schrader, Jürgen.

In: Circulation Research, Vol. 95, No. 8, 15.10.2004, p. 814-821.

Research output: Contribution to journalArticle

Koszalka, P, Özüyaman, B, Huo, Y, Zernecke, A, Flögel, U, Braun, N, Buchheiser, A, Decking, UKM, Smith, ML, Sévigny, J, Gear, A, Weber, AA, Molojavyi, A, Ding, Z, Weber, C, Ley, K, Zimmermann, H, Gödecke, A & Schrader, J 2004, 'Targeted disruption of cd73/ecto-5′-nucleotidase alters thromboregulation and augments vascular inflammatory response', Circulation Research, vol. 95, no. 8, pp. 814-821. https://doi.org/10.1161/01.RES.0000144796.82787.6f
Koszalka, Patrycja ; Özüyaman, Burcin ; Huo, Yuqing ; Zernecke, Alma ; Flögel, Ulrich ; Braun, Norbert ; Buchheiser, Anja ; Decking, Ulrich K.M. ; Smith, Michael L. ; Sévigny, Jean ; Gear, Adrian ; Weber, Artur Aron ; Molojavyi, Andrei ; Ding, Zhaoping ; Weber, Christian ; Ley, Klaus ; Zimmermann, Herbert ; Gödecke, Axel ; Schrader, Jürgen. / Targeted disruption of cd73/ecto-5′-nucleotidase alters thromboregulation and augments vascular inflammatory response. In: Circulation Research. 2004 ; Vol. 95, No. 8. pp. 814-821.
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AU - Zernecke, Alma

AU - Flögel, Ulrich

AU - Braun, Norbert

AU - Buchheiser, Anja

AU - Decking, Ulrich K.M.

AU - Smith, Michael L.

AU - Sévigny, Jean

AU - Gear, Adrian

AU - Weber, Artur Aron

AU - Molojavyi, Andrei

AU - Ding, Zhaoping

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