title: Targeted disruption of cd73/ecto-5'-nucleotidase alters thromboregulation and augments vascular inflammatory response.
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 micromol/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.
authors.affiliation: Department of Cardiovascular Physiology, Heinrich-Heine-University Duesseldorf, Duesseldorf, Germany.
authors.forename: Patrycja
authors.initials: P
authors.lastname: Koszalka
authors.forename: Burcin
authors.initials: B
authors.lastname: Ozüyaman
authors.forename: Yuqing
authors.initials: Y
authors.lastname: Huo
authors.forename: Alma
authors.initials: A
authors.lastname: Zernecke
authors.forename: Ulrich
authors.initials: U
authors.lastname: Flögel
authors.forename: Norbert
authors.initials: N
authors.lastname: Braun
authors.forename: Anja
authors.initials: A
authors.lastname: Buchheiser
authors.forename: Ulrich K M
authors.initials: UK
authors.lastname: Decking
authors.forename: Michael L
authors.initials: ML
authors.lastname: Smith
authors.forename: Jean
authors.initials: J
authors.lastname: Sévigny
authors.forename: Adrian
authors.initials: A
authors.lastname: Gear
authors.forename: Artur-Aron
authors.initials: AA
authors.lastname: Weber
authors.forename: Andrei
authors.initials: A
authors.lastname: Molojavyi
authors.forename: Zhaoping
authors.initials: Z
authors.lastname: Ding
authors.forename: Christian
authors.initials: C
authors.lastname: Weber
authors.forename: Klaus
authors.initials: K
authors.lastname: Ley
authors.forename: Herbert
authors.initials: H
authors.lastname: Zimmermann
authors.forename: Axel
authors.initials: A
authors.lastname: Gödecke
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schrader
mesh_terms: D015720:5'-Nucleotidase / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D000241:Adenosine / Q000096:biosynthesis / Q000502:physiology*; D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D001777:Blood Coagulation / Q000502:physiology; D002339:Carotid Arteries / Q000201:enzymology / Q000473:pathology; D002448:Cell Adhesion / Q000502:physiology; D002634:Chemotaxis, Leukocyte / Q000502:physiology; D003326:Coronary Circulation / Q000235:genetics; D003331:Coronary Vessels / Q000201:enzymology; D000242:Cyclic AMP / Q000097:blood; D004730:Endothelium, Vascular / Q000201:enzymology*; D045604:Extracellular Fluid / Q000201:enzymology; D005260:Female; D018390:Gene Targeting; D007249:Inflammation / Q000201:enzymology; D007511:Ischemia / Q000503:physiopathology; D007962:Leukocytes / Q000502:physiology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000098:blood supply; D017202:Myocardial Ischemia / Q000201:enzymology / Q000503:physiopathology; D015539:Platelet Activation / Q000502:physiology; D018047:Receptors, Purinergic P1 / Q000502:physiology; D015424:Reperfusion
chemical_list: D018047:Receptors, Purinergic P1; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D015720:5'-Nucleotidase; D000241:Adenosine; C002630:alpha,beta-methyleneadenosine 5'-triphosphate