Protein kinases A and C regulate receptor-mediated increases in cAMP in rabbit erythrocytes

Shaquria P. Adderley, Meera Sridharan, Elizabeth A. Bowles, Alan H. Stephenson, Mary L. Ellsworth, Randy S. Sprague
2010 American Journal of Physiology. Heart and Circulatory Physiology  
Adderley SP, Sridharan M, Bowles EA, Stephenson AH, Ellsworth ML, Sprague RS. Protein kinases A and C regulate receptormediated increases in cAMP in rabbit erythrocytes. Activation of the ␤-adrenergic receptor (␤-AR) or the prostacyclin receptor (IPR) results in increases in cAMP and ATP release from erythrocytes. cAMP levels depend on a balance between synthesis via adenylyl cyclase and hydrolysis by phosphodiesterases (PDEs). Previously, we reported that cAMP increases associated with
more » ... on of the ␤-AR and IPR in rabbit and human erythrocytes are tightly regulated by distinct PDEs (1). Importantly, inhibitors of these PDEs potentiated both increases in cAMP and ATP release. It has been shown that increases in protein kinase (PK) activity can activate PDE3 and PDE4. Both PKA and PKC are present in the erythrocyte and can phosphorylate and activate these PDEs. Here we investigate the hypothesis that PKA regulates PDE activity associated with the ␤-AR and both PKA and PKC regulate the PDE activity associated with the IPR in rabbit erythrocytes. Pretreatment of erythrocytes with the PKA inhibitor, H89 (10 M), in the presence of the PDE4 inhibitor, rolipram (10 M), augmented isoproterenol (1 M)-induced cAMP increases. In contrast, in the presence of the PDE3 inhibitor, cilostazol (10 M), pretreatment of erythrocytes with either H89 (1 M) or two chemically dissimilar inhibitors of PKC, calphostin C (1 M) or GFX109203X (1 M), potentiated iloprost (1 M)-induced cAMP increases. Furthermore, pretreatment of erythrocytes with both H89 and GFX109203X in the presence of cilostazol augmented the iloprost-induced increases in cAMP to a greater extent than either PK inhibitor individually. These results support the hypothesis that PDEs associated with receptor-mediated increases in cAMP in rabbit erythrocytes are regulated by kinases specific to the receptor's signaling pathway.
doi:10.1152/ajpheart.00975.2009 pmid:20008267 pmcid:PMC2822589 fatcat:zjvaa3bptfdlnfglxdlfjpc734