Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5
| Autor: | Kelly M. Thuet, Randy S. Sprague, Mary L. Ellsworth, Shaquria P. Adderley, Elizabeth A. Bowles, Meera Sridharan, Alan H. Stephenson |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Erythrocytes Purinones Phosphodiesterase Inhibitors Phosphodiesterase 3 Prostacyclin red blood cell Biology 03 medical and health sciences chemistry.chemical_compound Cytosol 0302 clinical medicine Clinical Research zaprinast Cyclic AMP medicine Animals Humans Receptor Cyclic GMP Vinca Alkaloids 030304 developmental biology Cyclic Nucleotide Phosphodiesterases Type 5 0303 health sciences isoproterenol Activator (genetics) Phosphodiesterase General Medicine Cyclic Nucleotide Phosphodiesterases Type 3 Cyclic Nucleotide Phosphodiesterases Type 4 Cell biology Isoenzymes cGMP chemistry Biochemistry Spermine Rabbits PDE5 PDE10A Soluble guanylyl cyclase Zaprinast 030217 neurology & neurosurgery medicine.drug |
| Zdroj: | Medical Science Monitor : International Medical Journal of Experimental and Clinical Research |
| ISSN: | 1234-1010 |
| DOI: | 10.12659/msm.881763 |
| Popis: | Summary 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. |
| Databáze: | OpenAIRE |
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