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    Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5

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    Author
    Adderley, Shaquria P.; Thuet, Kelly M.; Sridharan, Meera; Bowles, Elizabeth A.; Stephenson, Alan H.; Ellsworth, Mary L.; Sprague, Randy S.
    Abstract
    Background Within erythrocytes (RBCs), cAMP levels are regulated by phosphodiesterases (PDEs). Increases in cAMP and ATP release associated with activation of β-adrenergic receptors (βARs) and prostacyclin receptors (IPRs) are regulated by PDEs 2, 4 and PDE 3, respectively. Here we establish the presence of cytosolic PDEs in RBCs and determine a role for PDE5 in regulating levels of cGMP. Material/Methods Purified cytosolic proteins were obtained from isolated human RBCs and western analysis was performed using antibodies against PDEs 3A, 4 and 5. Rabbit RBCs were incubated with dbcGMP, a cGMP analog, to determine the effect of cGMP on cAMP levels. To determine if cGMP affects receptor-mediated increases in cAMP, rabbit RBCs were incubated with dbcGMP prior to addition of isoproterenol (ISO), a βAR receptor agonist. To demonstrate that endogenous cGMP produces the same effect, rabbit and human RBCs were incubated with SpNONOate (SpNO), a nitric oxide donor, and YC1, a direct activator of soluble guanylyl cyclase (sGC), in the absence and presence of a selective PDE5 inhibitor, zaprinast (ZAP). Results Western analysis identified PDEs 3A, 4D and 5A. dbcGMP produced a concentration dependent increase in cAMP and ISO-induced increases in cAMP were potentiated by dbcGMP. In addition, incubation with YC1 and SpNO in the presence of ZAP potentiated βAR-induced increases in cAMP. Conclusions PDEs 2, 3A and 5 are present in the cytosol of human RBCs. PDE5 activity in RBCs regulates cGMP levels. Increases in intracellular cGMP augment cAMP levels. These studies suggest a novel role for PDE5 in erythrocytes.
    URI
    http://hdl.handle.net/10342/5791
    Subject
     red blood cell; cGMP; isoproterenol; PDE5; zaprinast 
    Date
    2011
    Citation:
    APA:
    Adderley, Shaquria P., & Thuet, Kelly M., & Sridharan, Meera, & Bowles, Elizabeth A., & Stephenson, Alan H., & Ellsworth, Mary L., & Sprague, Randy S.. (January 2011). Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, (17:5), p.CR241-CR247. Retrieved from http://hdl.handle.net/10342/5791

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    MLA:
    Adderley, Shaquria P., and Thuet, Kelly M., and Sridharan, Meera, and Bowles, Elizabeth A., and Stephenson, Alan H., and Ellsworth, Mary L., and Sprague, Randy S.. "Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5". Medical Science Monitor : International Medical Journal of Experimental and Clinical Research. 17:5. (CR241-CR247.), January 2011. August 10, 2022. http://hdl.handle.net/10342/5791.
    Chicago:
    Adderley, Shaquria P. and Thuet, Kelly M. and Sridharan, Meera and Bowles, Elizabeth A. and Stephenson, Alan H. and Ellsworth, Mary L. and Sprague, Randy S., "Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5," Medical Science Monitor : International Medical Journal of Experimental and Clinical Research 17, no. 5 (January 2011), http://hdl.handle.net/10342/5791 (accessed August 10, 2022).
    AMA:
    Adderley, Shaquria P., Thuet, Kelly M., Sridharan, Meera, Bowles, Elizabeth A., Stephenson, Alan H., Ellsworth, Mary L., Sprague, Randy S.. Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research. January 2011; 17(5) CR241-CR247. http://hdl.handle.net/10342/5791. Accessed August 10, 2022.
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