eNOS phosphorylation and translocation are altered in male but not female mice by increased activation of the Gαq protein

2010 ◽  
Vol 88 (2) ◽  
pp. 121-129 ◽  
Author(s):  
C. Ruiz-Holst ◽  
B. Bölck ◽  
A. Ghanem ◽  
K. Tiemann ◽  
S. Brokat ◽  
...  
Keyword(s):  
Protein Expression ◽  
Cardiac Tissue ◽  
Weight Ratio ◽  
Wild Type ◽  
Mice Model ◽  
Enos Phosphorylation ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
Enos Protein Expression ◽  
Enos Protein

Little is known about sex-dependent physiological and pathophysiological differences in cardiac endothelial nitric oxide synthase (eNOS) expression and activation. Therefore, we investigated cardiac morphology and eNOS protein expression, including its translocation-dependent activation and phosphorylation, in cardiac tissue of male and female wild-type mice and transgenic heart-failure mice having a cardiac-specific, 5-fold overexpression of the Gαq protein. In addition, we measured calcineurin protein expression. Heart-to-body weight ratio was increased in Gαq mice. Female wild-type mice showed higher eNOS protein expression and activation (translocation and phosphorylation) than did wild-type males. In cardiac tissue of Gαq mice, these sex-dependent differences remained or were enhanced. Protein expression of the catalytic subunit calcineurin A, which has been shown to dephosphorylate eNOS, was higher in wild-type males than in wild-type females. These differences were increased in the Gαq mice model. We conclude that sex differences exist in cardiac eNOS protein expression and phosphorylation. Increased activation of the Gαq protein appears to alter eNOS protein expression and phosphorylation only in males.

Prolonged AMP-activated protein kinase induction impairs vascular functions

2013 ◽  
Vol 91 (12) ◽  
pp. 1025-1030 ◽  
Author(s):  
Saadet Turkseven ◽  
Elif Ertuna
Keyword(s):  
Protein Kinase ◽  
Protein Expression ◽  
Response Curves ◽  
Compound C ◽  
Amp Activated Protein Kinase ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
The Impact ◽  
Enos Protein Expression ◽  
Enos Protein

AMP-activated protein kinase (AMPK) is a regulator of cellular metabolism and is involved in the pathogenesis of several diseases, including type 2 diabetes and cardiovascular diseases. Data showing the effects of AMPK on vasculature are controversial. Therefore, the aim of this study was to determine the impact of prolonged AMPK activation on vascular functions. For this purpose we have examined the role of AMPK in endothelium-dependent and -independent relaxation and vascular contractions. For this, we incubated thoracic aortic rings, from rats, with AMPK activator 5-aminoimidazole-4-carboxamide-1-4-ribofuranoside (AICAR, 500 μmol/L or 2 mmol/L) in the presence or absence of AMPK inhibitor compound C (10 μmol/L). Next, cumulative dose–response curves to acetylcholine (ACh) (10−9−10−4 mol/L), nitroglycerine (NG) (10−9–3 × 10−5 mol/L), and noradrenaline (NA) (10−9−10−4 mol/L) were obtained. Endothelial nitric oxide synthase (eNOS) protein expression was determined. Our results show that endothelium-dependent relaxation was inhibited after AICAR treatment, and that this effect was reversed by AMPK inhibition. Moreover, AICAR enhanced the contractile response to NA and caused a decrease in eNOS protein expression. In conclusion, prolonged AMPK induction causes endothelial impairment, possibly via increased degradation and (or) reduced expression of eNOS.

NO from smooth muscle cells decreases NOS expression in endothelial cells: role of TNF-α

1999 ◽  
Vol 277 (4) ◽  
pp. H1317-H1325 ◽  
Author(s):  
Trinidad de Frutos ◽  
Lourdes Sánchez de Miguel ◽  
Margarita García-Durán ◽  
Fernando González-Fernández ◽  
Juan A. Rodríguez-Feo ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Smooth Muscle ◽  
Nitric Oxide Synthase ◽  
Protein Expression ◽  
Enos Expression ◽  
Tnf Α ◽  
Oxide Synthase ◽  
Enos Protein Expression ◽  
Enos Protein

Despite the evidence that cytokines stimulate nitric oxide (NO) production by inducible nitric oxide synthase (iNOS), several reports recently demonstrated that the hypotensive response related to endothelial nitric oxide synthase (eNOS) activity could be inhibited by the same cytokines. The aim of the present work was to analyze whether NO generated by vascular smooth muscle cells (VSMC) could modify eNOS protein expression in endothelial cells. Bovine aortic endothelial cells (BAEC) and bovine VSMC (BVSMC) in coculture were used for the study. Interleukin-1β (IL-1β, 10 ng/ml)-treated BVSMC, which expressed iNOS protein, decreased eNOS protein expression in BAEC. The presence of NO antagonists N ω-nitro-l-arginine methyl ester (10−3 mol/l) or N G-monomethyl-l-arginine (10−3 mol/l) prevented the decrease in eNOS protein expression induced by IL-1β-treated BVSMC. Surprisingly, two different NO donors, 3-morpholinosydnonimine (10−4 mol/l) and S-nitroso- N-acetyl-d,l-penicillamine (10−4 mol/l), failed to modify eNOS expression in BAEC, suggesting the existence of a diffusible mediator released from IL-1β-treated BVSMC that acts on endothelial cells by reducing eNOS expression. The presence of NO antagonists reduced tumor necrosis factor-α (TNF-α) production by IL-1β-stimulated BVSMC. This effect was also produced in the presence of a protein kinase G inhibitor, guanosine-5′-O-(2-thiodiphosphate) trilithium salt. A polyclonal antibody against TNF-α prevented eNOS expression in the BAEC-BVSMC coculture. In conclusion, NO by itself failed to modify eNOS protein expression in endothelial cells but increased TNF-α generation by IL-1β-stimulated BVSMC and, in this way, reduced eNOS expression in the endothelium.

scholarly journals Expression of endothelial nitric oxide synthase in the ovine ovary throughout the estrous cycle

Reproduction ◽  
2006 ◽  
Vol 132 (4) ◽  
pp. 579-587 ◽  
Author(s):  
Anna T Grazul-Bilska ◽  
Chainarong Navanukraw ◽  
Mary Lynn Johnson ◽  
Daniel A Arnold ◽  
Lawrence P Reynolds ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Blood Vessels ◽  
Estrous Cycle ◽  
Endothelial Nitric Oxide Synthase ◽  
Follicular Growth ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
Enos Protein Expression ◽  
Enos Protein

This study was conducted to evaluate the expression of endothelial nitric oxide synthase (eNOS) in ovarian follicles and corpora lutea (CL) throughout the estrous cycle in sheep. Three experiments were conducted to (1) immunolocalize eNOS protein, (2) determine expression of mRNA for eNOS and its receptor guanylate cyclase 1 soluble β3 (GUCY1B3), and (3) co-localize eNOS and vascular endothelial growth factor (VEGF) proteins in the follicles and/or CL throughout the estrous cycle. In experiment 1, ovaries were collected from ewes treated with FSH, to induce follicular growth or atresia. In experiment 2, ovaries were collected from ewes treated with FSH and hCG to induce follicular growth and ovulation. In experiment 3, ovaries were collected from superovulated ewes to generate multiple CL on days 2, 4, 10, and 15 of the estrous cycle. In experiments 1 and 2, the expression of eNOS protein was detected in the blood vessels of the theca externa and interna of healthy ovarian follicles. However, in early and advanced atretic follicles, eNOS protein expression was absent or reduced. During the immediate postovulatory period, eNOS protein expression was detected in thecal-derived cells that appeared to be invading the granulosa layer. Expression of eNOS mRNA tended to increase in granulosa cells at 12 and 24 h, and in theca cells 48 h after hCG injection. In experiment 3, eNOS protein was located in the blood vessels of the CL during the estrous cycle. Dual localization of eNOS and VEGF proteins in the CL demonstrated that both were found in the blood vessels.

scholarly journals Developmental changes in endothelial nitric oxide synthase expression and activity in ovine fetal lung

2000 ◽  
Vol 278 (1) ◽  
pp. L202-L208 ◽  
Author(s):  
Thomas A. Parker ◽  
Timothy D. le Cras ◽  
John P. Kinsella ◽  
Steven H. Abman
Keyword(s):  
Nitric Oxide ◽  
Protein Expression ◽  
Fetal Lung ◽  
Late Gestation ◽  
Enos Expression ◽  
Enos Mrna ◽  
Ovine Fetal ◽  
Endothelial Nitric Oxide ◽  
Enos Protein Expression ◽  
Enos Protein

Endothelial nitric oxide (NO) synthase (eNOS) produces NO, which contributes to vascular reactivity in the fetal lung. Pulmonary vasoreactivity develops during late gestation in the ovine fetal lung, during the period of rapid capillary and alveolar growth. Although eNOS expression peaks near birth in the fetal rat, lung capillary and distal air space development occur much later than in the fetal lamb. To determine whether lung eNOS expression in the lamb differs from the timing and pattern reported in the rat, we measured eNOS mRNA and protein by Northern and Western blot analyses and NOS activity by the arginine-to-citrulline conversion assay in lung tissue from fetal, newborn, and maternal sheep. Cellular localization of eNOS expression was determined by immunohistochemistry. eNOS mRNA, protein, and activity were detected in samples from all ages, and eNOS was expressed predominantly in the vascular endothelium. Lung eNOS mRNA expression increases from low levels at 70 days gestation to peak at 113 days and remains high for the rest of fetal life. Newborn eNOS mRNA expression does not change from fetal levels but is lower in the adult ewe. Lung eNOS protein expression in the fetus rises and peaks at 118 days gestation but decreases before birth. eNOS protein expression rises in the newborn period but is lower in the adult. Lung NOS activity also peaks at 118 days gestation in the fetus before falling in late gestation and remaining low in the newborn and adult. We conclude that the pattern of lung eNOS expression in the sheep differs from that in the rat and may reflect species-related differences in lung development. We speculate that the rise in fetal lung eNOS may contribute to the marked lung growth and angiogenesis that occurs during the same period of time.

Developmental expression of eNOS in postnatal swine mesenteric artery

2002 ◽  
Vol 283 (6) ◽  
pp. G1328-G1335 ◽  
Author(s):  
Kristina M. Reber ◽  
Baogen Y. Su ◽  
K. Reed Clark ◽  
Dana L. Pohlman ◽  
Charles E. Miller ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mesenteric Artery ◽  
Developmental Expression ◽  
Early Postnatal Development ◽  
Before And After ◽  
Enos Mrna ◽  
Group Resistance ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide ◽  
Enos Protein

Developmental changes in the expression of endothelial nitric oxide synthase (eNOS) within the mesenteric artery of swine were studied in fetal (110 days postconception/117 days total gestation) and on postnatal days 1, 3, 10, and 30. Subjects in the 1-day-old group were subdivided into fed and nonfed. Transcription of eNOS was determined by real-time PCR, protein expression was evaluated by Western blotting, and hemodynamic and oxygenation parameters were measured within in situ gut loops before and after the administration of N G-monomethyl-l-arginine (l-NMMA). The abundance of eNOS mRNA remained steady throughout all ages. In contrast, expression of eNOS protein was twofold greater in the 1-day-old fed subjects compared with fetal or 1-day-old nonfed subjects. eNOS protein expression remained elevated on day 3, increased on day 10, and then declined to a level similar to the day 1 nonfed group by postnatal day 30. Intestinal vascular resistance was 31% lower in the day 1 fed group when compared to the day 1 nonfed group; resistance continued to decline through day 10 but then significantly increased on day 30. We conclude that the expression of eNOS changes within the mesenteric artery during early postnatal development at a posttranscriptional level.

Cotyledon and binucleate cell nitric oxide synthase expression in an ovine model of fetal growth restriction

2001 ◽  
Vol 90 (6) ◽  
pp. 2420-2426 ◽  
Author(s):  
Henry L. Galan ◽  
Timothy R. H. Regnault ◽  
Timothy D. Le Cras ◽  
R. Weslie Tyson ◽  
Russell V. Anthony ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Protein Expression ◽  
Protein Content ◽  
Vascular Endothelium ◽  
Heat Exposure ◽  
Growth Restriction ◽  
Oxide Synthase ◽  
Enos Protein Expression ◽  
Enos Protein

Heat exposure early in ovine pregnancy results in placental insufficiency and intrauterine growth restriction (PI-IUGR). We hypothesized that heat exposure in this model disrupts placental structure and reduces placental endothelial nitric oxide synthase (eNOS) protein expression. We measured eNOS protein content and performed immunohistochemistry for eNOS in placentas from thermoneutral (TN) and hyperthermic (HT) animals killed at midgestation (90 days). Placental histomorphometry was compared between groups. Compared with the TN controls, the HT group showed reduced delivery weights (457 ± 49 vs. 631 ± 21 g; P < 0.05) and a trend for reduced placentome weights (288 ± 61 vs. 554 ± 122 g; P = 0.09). Cotyledon eNOS protein content was reduced by 50% in the HT group ( P < 0.03). eNOS localized similarly to the vascular endothelium and binucleated cells (BNCs) within the trophoblast of both experimental groups. HT cotyledons showed a reduction in the ratio of fetal to maternal stromal tissue (1.36 ± 0.36 vs. 3.59 ± 1.2; P≤ 0.03). We conclude that eNOS protein expression is reduced in this model of PI-IUGR and that eNOS localizes to both vascular endothelium and the BNC. We speculate that disruption of normal vascular development and BNC eNOS production and function leads to abnormal placental vascular tone and blood flow in this model of PI-IUGR.

Endothelial nitric oxide synthase increases in left atria of dogs with pacing-induced heart failure

1998 ◽  
Vol 275 (6) ◽  
pp. H1971-H1978 ◽  
Author(s):  
Fadi H. Khadour ◽  
Darryl W. O’Brien ◽  
Yuling Fu ◽  
Paul W. Armstrong ◽  
Richard Schulz
Keyword(s):  
Heart Failure ◽  
Cardiac Tissue ◽  
Left Ventricular ◽  
No Production ◽  
Ventricular Activity ◽  
Plasma Nitrate ◽  
Oxide Synthase ◽  
Nos Isoforms ◽  
Endothelial Nitric Oxide ◽  
Enos Protein

In congestive heart failure (CHF) the alterations in cardiac NO synthase (NOS) isoforms activity and expression are incompletely documented and the chamber specificity of these changes is unknown. We studied plasma nitrate-nitrite ([Formula: see text]), atrial, and ventricular NOS activities and protein expression (Western blot and densitometric analysis) in nonpaced control dogs and in dogs paced for 2 or 21 days into CHF. Plasma [Formula: see text] rose significantly after 7 and 21 days of pacing, whereas creatinine levels remained unchanged. In control dogs Ca2+-dependent NOS activity in left atria was double that of right or left ventricular activity. In paced animals the activity increased only in the atria after 21 but not 2 days of pacing. Levels of endothelial NOS (eNOS) protein were enhanced in the left atria but not ventricles after 21 days of pacing because of a greater quantity of the 150-kDa but not the 135-kDa eNOS. Ca2+-independent NOS activity was undetectable in any cardiac tissue. The specific upregulation of eNOS in the left atria suggests that NO production may be enhanced to counterbalance hypertrophy that develops during pacing-induced CHF.

scholarly journals Ablation of eNOS does not promote adipose tissue inflammation

2016 ◽  
Vol 310 (8) ◽  
pp. R744-R751 ◽  
Author(s):  
Thomas J. Jurrissen ◽  
Ryan D. Sheldon ◽  
Michelle L. Gastecki ◽  
Makenzie L. Woodford ◽  
Terese M. Zidon ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Adipose Tissue ◽  
Control Diet ◽  
Western Diet ◽  
No Production ◽  
Wild Type ◽  
Oxphos Complex ◽  
Enos Phosphorylation ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Adipose tissue (AT) inflammation is a hallmark characteristic of obesity and an important determinant of insulin resistance and cardiovascular disease; therefore, a better understanding of factors regulating AT inflammation is critical. It is well established that reduced vascular endothelial nitric oxide (NO) bioavailability promotes arterial inflammation; however, the role of NO in modulating inflammation in AT remains disputed. In the present study, 10-wk-old C57BL6 wild-type and endothelial nitric oxide synthase (eNOS) knockout male mice were randomized to either a control diet (10% kcal from fat) or a Western diet (44.9% kcal from fat, 17% sucrose, and 1% cholesterol) for 18 wk ( n = 7 or 8/group). In wild-type mice, Western diet-induced obesity led to increased visceral white AT expression of inflammatory genes (e.g., MCP1, TNF-α, and CCL5 mRNAs) and markers of macrophage infiltration (e.g., CD68, ITGAM, EMR1, CD11C mRNAs, and Mac-2 protein), as well as reduced markers of mitochondrial content (e.g., OXPHOS complex I and IV protein). Unexpectedly, these effects of Western diet on visceral white AT were not accompanied by decreases in eNOS phosphorylation at Ser-1177 or increases in eNOS phosphorylation at Thr-495. Also counter to expectations, eNOS knockout mice, independent of the diet, were leaner and did not exhibit greater white or brown AT inflammation compared with wild-type mice. Collectively, these findings do not support the hypothesis that reduced NO production from eNOS contributes to obesity-related AT inflammation.

Increased cardiac microvascular permeability and activation of cardiac endothelial nitric oxide synthase in high tidal volume ventilation-induced lung injury

2010 ◽  
Vol 4 (1) ◽  
pp. 27-36
Author(s):  
Ming-Jui Hung ◽  
Ming-Yow Hung ◽  
Wen-Jin Cherng ◽  
Li-Fu Li
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Lung Injury ◽  
Tidal Volume ◽  
Endothelial Nitric Oxide Synthase ◽  
Microvascular Permeability ◽  
Akt Phosphorylation ◽  
Enos Phosphorylation ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Abstract Background: Positive pressure ventilation with large tidal volumes has been shown to cause lung injury via the serine/threonine kinase-protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS)-pathways. However, the effects of high tidal volume (VT) ventilation on the heart are unclear. Objectives: Evaluate the effect of VT ventilation on the cardiac vascular permeability and intracellular Akt and eNOS signaling pathway. Methods: C57BL/6 and Akt knock-out (heterozygotes, +/−) mice were exposed to high VT (30 mL/kg) mechanical ventilation with room air for one and/or five hours. Results: High VT ventilation increased cardiac microvascular permeability and eNOS phosphorylation in a timedependent manner. Serum cardiac troponin I was increased after one hour of high VT ventilation. Cardiac Akt phosphorylation was accentuated after one hour and attenuated after five hours of high VT ventilation. Pharmacological inhibition of Akt with LY294002 and high VT ventilation of Akt+/− mice attenuated cardiac Akt phosphorylation, but not eNOS phosphorylation. Conclusion: High VT ventilation increased cardiac myocardial injury, microvascular permeability, and eNOS phosphorylation. Involvement of cardiac Akt in high VT ventilation was transient.

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