title: Role of Purinergic Receptor Expression and Function for Reduced Responsiveness to Adenosine Diphosphate in Washed Human Platelets.
abstract: BACKGROUND
  Washing of platelets is an important procedure commonly used for experimental studies, e.g. in cardiovascular research. As a known phenomenon, responsiveness to adenosine diphosphate (ADP) is reduced in washed platelets, although underlying molecular mechanisms-potentially interfering with experimental results-have not been thoroughly studied.
  
  
  OBJECTIVE
  Since ADP mediates its effects via three purinergic receptors P2Y1, P2X1 and P2Y12, their surface expression and function were investigated in washed platelets and, for comparison, in platelet-rich-plasma (PRP) at different time points for up to 2 hours after preparation.
  
  
  RESULTS
  In contrast to PRP, flow cytometric analysis of surface expression in washed platelets revealed an increase of all receptors during the first 60 minutes after preparation followed by a significant reduction, which points to an initial preactivation of platelets and consecutive degeneration. The activity of the P2X1 receptor (measured by selectively induced calcium flux) was substantially maintained in both PRP and washed platelets. P2Y12 function (determined by flow cytometry as platelet reactivity index) was partially reduced after platelet washing compared to PRP, but remained stable in course of ongoing storage. However, the function of the P2Y1 receptor (measured by selectively induced calcium flux) continuously declined after preparation of washed platelets.
  
  
  CONCLUSIONS
  In conclusion, decreasing ADP responsiveness in washed platelets is particularly caused by impaired activity of the P2Y1 receptor associated with disturbed calcium regulation, which has to be considered in the design of experimental studies addressing ADP mediated platelet function.
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Juergen
authors.initials: J
authors.lastname: Koessler
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hermann
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Weber
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Angela
authors.initials: A
authors.lastname: Koessler
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Kuhn
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Boeck
authors.affiliation: Institute of Transfusion Medicine and Haemotherapy, University of Wuerzburg, Wuerzburg, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Kobsar
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects*; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects; D053657:Platelet-Rich Plasma / Q000166:cytology; D011983:Receptors, Purinergic / Q000378:metabolism*; D058474:Receptors, Purinergic P2X1 / Q000378:metabolism; D058472:Receptors, Purinergic P2Y1 / Q000378:metabolism; D058925:Receptors, Purinergic P2Y12 / Q000378:metabolism
chemical_list: D011983:Receptors, Purinergic; D058474:Receptors, Purinergic P2X1; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D000244:Adenosine Diphosphate
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