Nitric oxide-evoked transient kinetics of cyclic GMP in vascular smooth muscle cells
| Autor: | Carolyn L. Sawyer, Kara F. Brunelle, Sharon M Cawley, Albert van der Vliet, Wolfgang R. Dostmann |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Male
medicine.medical_specialty Vascular smooth muscle Phosphodiesterase 3 Kinetics Myocytes Smooth Muscle Biology Nitric Oxide Transfection Muscle Smooth Vascular Nitric oxide Rats Sprague-Dawley chemistry.chemical_compound Desensitization (telecommunications) 3' 5'-Cyclic-GMP Phosphodiesterases Internal medicine medicine Fluorescence Resonance Energy Transfer Animals Cyclic GMP Cells Cultured Cyclic Nucleotide Phosphodiesterases Type 5 Cell Biology PDE5 drug design Rats Endocrinology chemistry Guanylate Cyclase Biophysics Phosphorylation Indicators and Reagents PDE10A Signal Transduction |
| Zdroj: | Cellular signalling. 19(5) |
| ISSN: | 0898-6568 |
| Popis: | Cyclic-3′,5′-guanosine monophosphate (cGMP) mediates the intracellular signaling cascade responsible for the nitric oxide (NO) initiated relaxation of vascular smooth muscle (VSM). However, the temporal dynamics, including the regulation of cGMP turnover, are largely unknown. Here we report new mechanistic insights into the kinetics of cGMP synthesis and hydrolysis in primary VSM cells by utilizing FRET-based cGMP-indicators [A. Honda, S.R. Adams, C.L. Sawyer, V. Lev-Ram, R.Y. Tsien, W.R. Dostmann, Proc. Natl. Acad. Sci. U S A 98 (5) (2001) 2437.]. First, 2-( N , N -Diethylamino)-diazenolate 2-oxide (DEA/NO) and 2,2′-(Hydroxynitrosohydrazono)-bis-ethanimine (DETA/NO) induced NO-concentration dependent, transient cGMP responses (“peaks”) irrespective of their rates of NO release. The kinetic characteristics of these cGMP peaks were governed by the concerted action of the NO-sensitive guanylyl cyclase (GC) and phosphodiesterase type V (PDE5) as shown by their respective inhibition using 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and Sildenafil. These responses occurred in the presence of moderately elevated cGMP (5–15% FRET ratio), and thus activated PKG and phosphorylated PDE5, suggesting a prominent role for GC in the maintenance and termination of cGMP peaks. Furthermore, cGMP transients could be elicited repeatedly without apparent desensitization of GC or by suppression of cGMP via long-term PDE5 activity. These results demonstrate a continuous sensitivity of the NO/cGMP signaling system, inherent to the phasic nature of smooth muscle physiology. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect