Embryonic motor neuron dendrite growth is stunted by inhibition of nitric oxide-dependent activation of soluble guanylyl cyclase and protein kinase G

Atherosclerotic lower extremity peripheral artery disease (PAD) is among the most prevalent, morbid and mortal of all cardiovascular disorders. Pathologic arterial smooth muscle (ASM) cell migration is a major component of atherogenic PAD and efforts aimed at attenuating its progression are clinically essential. Cyclic nucleotide signaling has long been studied for its growth-mitigating properties in the setting of PAD and other vascular growth disorders. In this study we hypothesized that the novel, heme-independent soluble guanylyl cyclase activator BAY 60-2770 (BAY) inhibits ASM cell migration through phosphorylation of the protein kinase G (PKG) target and actin-binding protein vasodilator-stimulated phosphoprotein (VASP). In a rat model of injury-induced arterial growth, BAY significantly reduced neointima formation and luminal narrowing compared to vehicle (Veh)-treated control arteries after 2 weeks. Using rat and human ASM cells BAY significantly attenuated cell migration, reduced G:F actin, and increased cyclic GMP content, PKG activity and phosphorylated VASP at Ser239 (pVASP.S239) compared to Veh controls. Using site-directed mutagenesis, both full-length VASP-overexpressing (wild type, WT) and VASP.S239 phosphorylation-resistant mutants showed significantly reduced cell migration compared to naïve controls, however, there was no effect on cell migration between either VASP transfected group in the presence of BAY. Interestingly, both VASP mutants showed significantly increased PKG activity compared to naïve cells, and in turn pharmacologic PKG blockade in the presence of BAY fully reversed the inhibitory effect of BAY alone on cell migration. These data suggest BAY has capacity to inhibit ASM cell migration through cyclic GMP/PKG/VASP signaling yet through mechanisms independent of pVASP.S239. Findings from this study implicate BAY via cyclic GMP/PKG/VASP as a potential pharmacotherapeutic agent against aberrant ASM growth disorders such as PAD.

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Protein kinase G phosphorylates soluble guanylyl cyclase and inhibits its activity

BMC Pharmacology ◽

10.1186/1471-2210-7-s1-p45 ◽

2007 ◽

Vol 7 (S1) ◽

Author(s):

Andreas Papapetropoulos ◽

Zongmin Zhou ◽

Nazish Sayed ◽

Anastasia Pyriochou ◽

Charis Roussos ◽

...

Keyword(s):

Protein Kinase ◽

Guanylyl Cyclase ◽

Soluble Guanylyl Cyclase ◽

Protein Kinase G

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Preservation of nitric oxide-induced relaxation of porcine coronary artery: roles of the dimers of soluble guanylyl cyclase, phosphodiesterase type 5, and cGMP-dependent protein kinase

Pflügers Archiv - European Journal of Physiology ◽

10.1007/s00424-014-1441-2 ◽

2014 ◽

Vol 466 (10) ◽

pp. 1999-2008 ◽

Cited By ~ 4

Author(s):

Juan Liu ◽

Zhengju Chen ◽

Liping Ye ◽

Huixia Liu ◽

Dou Dou ◽

...

Keyword(s):

Nitric Oxide ◽

Coronary Artery ◽

Protein Kinase ◽

Guanylyl Cyclase ◽

Soluble Guanylyl Cyclase ◽

Dependent Protein Kinase ◽

Porcine Coronary Artery ◽

Cgmp Dependent Protein Kinase ◽

Dependent Protein ◽

Phosphodiesterase Type

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Gaseous Transmitter Regulation of Catecholamine Secretion by hypoxia in Murine Adrenal Chromaffin Cells

Journal of Neurophysiology ◽

10.1152/jn.00669.2020 ◽

2021 ◽

Author(s):

Anna Gridina ◽

Xiaoyu Su ◽

Shakil A. Khan ◽

Ying-Jie Peng ◽

Benjamin L Wang ◽

...

Keyword(s):

Nervous System ◽

Protein Kinase ◽

Guanylyl Cyclase ◽

Chromaffin Cells ◽

Soluble Guanylyl Cyclase ◽

Protein Kinase G ◽

Adrenal Chromaffin Cells ◽

Wild Type ◽

Genetic Deletion ◽

Adrenal Chromaffin

Emerging evidence suggests that gaseous molecules, carbon monoxide (CO) and hydrogen sulfide (H2S) generated by heme oxygenase-(HO)-2 and cystathionine γ-lyase (CSE), respectively, function as transmitters in the nervous system. Present study examined the roles of CO and H2S in hypoxia-induced catecholamine (CA) release from adrenal medullary chromaffin cells (AMC). Studies were performed on AMC from adult (≥6 weeks of age) wild type (WT), HO-2 null, CSE null and HO-2/CSE double null mice of either gender. CA secretion was determined by carbon fiber amperometry and [Ca2+]i by microflurometry using Fura-2. HO-2- and CSE immunoreactivities were seen in WT AMC, which were absent in HO-2 and CSE null mice. Hypoxia (medium pO2 30-38 mmHg) evoked CA release and elevated [Ca2+]i. The magnitude of hypoxic response was greater in HO-2 null mice and in HO inhibitor treated WT AMC compared to controls. H2S levels were elevated in HO-2 null AMC. Either pharmacological inhibition or genetic deletion of CSE prevented the augmented hypoxic responses of HO-2 null AMC and H2S donor rescued AMC responses to hypoxia in HO-2/CSE double null mice. CORM-3, a CO donor, prevented the augmented hypoxic responses in WT and HO-2 null AMC. CO donor reduced H2S levels in WT AMC. The effects of CO donor were blocked by either ODQ or 8pCT, inhibitors of soluble guanylyl cyclase (SGC) or protein kinase G, respectively. These results suggest that HO-2-derived CO inhibits hypoxia-evoked CA secretion from adult murine AMC involving soluble guanylyl cyclase (SGC)-protein kinase G (PKG)-dependent regulation of CSE- derived H2S.

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Endothelium-derived NO, but not cyclic GMP, is required for hypoxic augmentation in isolated porcine coronary arteries

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00258.2011 ◽

2011 ◽

Vol 301 (6) ◽

pp. H2313-H2321 ◽

Cited By ~ 29

Author(s):

Calvin K. Y. Chan ◽

Judith Mak ◽

Yuansheung Gao ◽

Ricky Y. K. Man ◽

Paul M. Vanhoutte

Keyword(s):

Nitric Oxide ◽

Protein Kinase ◽

Coronary Arteries ◽

Cyclic Gmp ◽

Guanylyl Cyclase ◽

Calcium Influx ◽

Soluble Guanylyl Cyclase ◽

Isometric Tension ◽

Nitric Oxide Donor ◽

Atpase Inhibitor

The present study investigated the mechanism underlying the transient potentiation of vasoconstriction by hypoxia in isolated porcine coronary arteries. Isometric tension was measured in rings with or without endothelium. Hypoxia (Po2 <30 mmHg) caused a transient further increase in tension (hypoxic augmentation) in contracted (with U46619) preparations. The hypoxic response was endothelium dependent and abolished by inhibitors of nitric oxide synthase [ Nω-nitro-l-arginine methyl ester (l-NAME)] or soluble guanylyl cyclase (ODQ and NS2028). The addition of DETA NONOate (nitric oxide donor) in the presence of l-NAME restored the hypoxic augmentation, suggesting the involvement of the nitric oxide pathway. However, the same was not observed after incubation with 8-bromo-cyclic GMP, atrial natriuretic peptide, or isoproterenol. Assay of the cyclic GMP content showed no change upon exposure to hypoxia in preparations with and without endothelium. Incubation with protein kinase G and protein kinase A inhibitors did not inhibit the hypoxic augmentation. Thus the hypoxic augmentation is dependent on nitric oxide and soluble guanylyl cyclase but independent of cyclic GMP. The hypoxic augmentation persisted in calcium-free buffer and in the presence of nifedipine, ruling out a role for extracellular calcium influx. Hypoxia did not alter the intracellular calcium concentration, as measured by confocal fluorescence microscopy. This observation and the findings that hypoxic augmentation is enhanced by thapsigargin (sarco/endoplasmic reticulum calcium ATPase inhibitor) and inhibited by HA1077 or Y27632 (Rho kinase inhibitors) demonstrate the involvement of calcium sensitization in the phenomenon.

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Differential Contribution of the Guanylyl Cyclase-Cyclic GMP-Protein Kinase G Pathway to the Proliferation of Neural Stem Cells Stimulated by Nitric Oxide

Neurosignals ◽

10.1159/000332811 ◽

2013 ◽

Vol 21 (1-2) ◽

pp. 1-13 ◽

Cited By ~ 16

Author(s):

Bruno P. Carreira ◽

Maria Inês Morte ◽

Ana Sofia Lourenço ◽

Ana Isabel Santos ◽

Ângela Inácio ◽

...

Keyword(s):

Nitric Oxide ◽

Stem Cells ◽

Protein Kinase ◽

Neural Stem Cells ◽

Cyclic Gmp ◽

Guanylyl Cyclase ◽

Protein Kinase G ◽

Differential Contribution

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Activation of the nitric oxide-cGMP pathway reduces phasic contractions in neonatal rat bladder strips via protein kinase G

AJP Renal Physiology ◽

10.1152/ajprenal.00207.2009 ◽

2009 ◽

Vol 297 (2) ◽

pp. F333-F340 ◽

Cited By ~ 20

Author(s):

Debra E. Artim ◽

F. Aura Kullmann ◽

Stephanie L. Daugherty ◽

Hsi-Yang Wu ◽

William C. de Groat

Keyword(s):

Nitric Oxide ◽

Protein Kinase ◽

Soluble Guanylyl Cyclase ◽

Inhibitory Effect ◽

Neonatal Rat ◽

Protein Kinase G ◽

No Donor ◽

Adult Rat ◽

Rat Bladder ◽

Normal Bladder

Nitric oxide (NO), a neurotransmitter in the lower urinary tract, stimulates soluble guanylyl cyclase (sGC) and in turn cGMP-dependent protein kinase G (PKG) to modulate a number of downstream targets. NO donors reduce bladder hyperactivity in some pathological models but do not affect normal bladder activity in the adult rat. In this study, the NO donor S-nitroso- N-acetyl-dl-penicillamine (SNAP; 100 μM) decreased the amplitude and frequency of spontaneous and carbachol-enhanced contractions in neonatal rat bladder strips, which are intrinsically hyperactive. This effect was blocked by inhibition of sGC and mimicked by application of a membrane-permeable cGMP analog (8-bromo-cGMP, 100 μM). Inhibition of PKG prevented or reversed the inhibitory effects of 8-bromo-cGMP. A portion of the SNAP-mediated inhibition was also dependent upon PKG; however, a short-lasting, sGC-dependent inhibitory effect of SNAP was still present after PKG inhibition. Inhibition of NO synthase with l-NAME (100 μM) did not change the amplitude or frequency of contractions. However, inhibition of endogenous phosphodiesterase (PDE)-5 with zaprinast (25 μM) reduced the amplitude and frequency of phasic contractions and increased the magnitude of inhibition produced by maximal concentrations of SNAP, suggesting that endogenous PDEs are constitutively active and regulate cGMP production. These results suggest that the NO-cGMP-PKG pathway may be involved in inhibitory control of the neonatal rat bladder.

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