Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins.
AuthorsKojima H, Nakatsubo N, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Nagano T
JournalAnal Chem
PubMed ID9666719
Nitric oxide is a gaseous, free radical which plays a role as an intracellular second messenger and a diffusable intercellular messenger. To obtain direct evidence for NO functions in vivo, we have designed and synthesized diaminofluoresceins (DAFs) as novel fluorescent indicators for NO. The fluorescent chemical transformation of DAFs is ... More
Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA.
AuthorsKojima H, Nakatsubo N, Kikuchi K, Urano Y, Higuchi T, Tanaka J, Kudo Y, Nagano T
JournalNeuroreport
PubMed ID9855277
The biological functions of nitric oxide in the neuronal system remain controversial. Using a novel fluorescence indicator, DAF-2 DA, for direct detection of NO, we examined both acute rat brain slices and organotypic culture of brain slices to ascertain NO production sites. The fluorescence intensity in the CA1 region of ... More
Development of a fluorescent indicator for nitric oxide based on the fluorescein chromophore.
AuthorsKojima H, Sakurai K, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Nagano T
JournalChem Pharm Bull (Tokyo)
PubMed ID9501473
Endogenous nitric oxide (NO) appears to modulate many physiological and pathophysiological processes. In order to obtain direct evidence for NO functions in vivo, we have developed 4,5-diaminofluorescein (DAF-2) as a novel fluorescent indicator for NO. Green-fluorescent triazolofluorescein formed by the reaction of NO and DAF-2 affords high sensitivity for NO ... More
Increased nitric oxide-dependent nitrosylation of 4,5-diaminofluorescein by oxidants: implications for the measurement of intracellular nitric oxide.
AuthorsJourd'heuil D
JournalFree Radic Biol Med
PubMed ID12208354
4,5 diaminofluorescein (DAF-2) is increasingly utilized as a fluorescent detector for nitric oxide (*NO) in cells and tissues. In oxygenated solutions, reactive nitrogen species derived from (*) NO autoxidation nitrosate DAF-2 to yield the highly fluorescent DAF-2 triazole. In the present study, we investigated the nitrosation of DAF-2 at a ... More
Increased basal nitric oxide release despite enhanced free radical production in hypertension.
AuthorsMaffei A, Poulet R, Vecchione C, Colella S, Fratta L, Frati G, Trimarco V, Trimarco B, Lembo G
JournalJ Hypertens
PubMed ID12023683
INTRODUCTION : Although in hypertension a defect in stimulated nitric oxide (NO) is well established, little is known about basal NO levels. Thus, we measured directly in vessels from normotensive [Wistar-Kyoto (WKY)] rats and spontaneously hypertensive rats (SHR) both basal and stimulated NO production using a novel technique [4,5-diaminofluorescein (DAF-2) ... More
4,5-diaminofluoroscein imaging of nitric oxide synthesis in crayfish terminal ganglia.
AuthorsSchuppe H, Cuttle M, Chad JE, Newland PL
JournalJ Neurobiol
PubMed ID12382263
We have analyzed the synthesis of nitric oxide in the terminal abdominal ganglion of the crayfish using the fluorescent probe 4,5-Diaminofluoroscein diacetate, DAF-2 DA. Following DAF-2 loading, ganglia showed cell-specific patterns of fluorescence in which the occurrence of strongly fluorescent cell bodies was highest in specific anterior, central, and posterior ... More
Nitric oxide synthase in filariae: demonstration of nitric oxide production by embryos in Brugia malayi and Acanthocheilonema viteae.
AuthorsPfarr KM, Qazi S, Fuhrman JA
JournalExp Parasitol
PubMed ID11384164
The radical gas nitric oxide (NO) is synthesized by nitric oxide synthase (NOS) from l-arginine and molecular oxygen. Nitric oxide is an important signaling molecule in invertebrate and vertebrate systems. Previously we have shown that NOS is localized to more tissues in Brugia malayi than has been reported in Ascaris ... More
Endocrinology: nitric oxide-mediated insulin secretion in response to citrulline in islet beta-cells.
AuthorsNakata M, Yada T
JournalPancreas
PubMed ID14508123
INTRODUCTION: Nitric oxide (NO) synthases (NOSs) are expressed in insulin secreting beta-cells. However, physiologic role of NO in insulin release is still controversial. We previously reported that argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL), together with NOS, constitute the citrulline-argininosuccinate-arginine (Cit-AS-Arg) cycle in beta-cells and that this cycle metabolizes citrulline ... More
The demonstration of nitric oxide in cochlear blood vessels in vivo and in vitro: the role of endothelial nitric oxide in venular permeability.
AuthorsShi X, Nuttall AL
JournalHear Res
PubMed ID12361868
The objectives of the current study were to investigate the distribution and production of NO in cochlear blood vessels, and to assess whether the inhibition of basal NO production leads to vascular protein leakage of the cochlear microvasculature. Using the fluorescent dye 4,5-diaminofluorescein diacetate, NO was detected, both in vitro ... More
Serum nitric oxide (NO) levels in systemic sclerosis patients: correlation between NO levels and clinical features.
AuthorsTakagi K, Kawaguchi Y, Hara M, Sugiura T, Harigai M, Kamatani N
JournalClin Exp Immunol
PubMed ID14632763
Vascular damage in systemic sclerosis (SSc) may be a factor in the abnormal regulation of several vasoactive agents. It has been well confirmed that plasma endothelin-1, as a vasoconstrictive factor, is elevated in patients with SSc. However, it is still controversial whether the levels of serum nitric oxide (NO), a ... More
NMDA induces NOS 1 translocation to the cell membrane in NGF-differentiated PC 12 cells.
AuthorsArundine M, Sanelli T, Ping He B, Strong MJ
JournalBrain Res
PubMed ID12763249
Glutamatergic-mediated nitric oxide (NO) production occurs via the N-methyl-D-aspartic acid (NMDA) postsynaptic density protein 95 (PSD95)-neuronal nitric oxide synthase (NOS1) ternary complex. To determine whether NOS1 is targeted to the membrane subsequent to NMDA receptor activation, we examined the effect of NMDA on NOS1 subcellular localization in nerve growth factor ... More
Insulin receptor substrate-1 and phosphoinositide-dependent kinase-1 are required for insulin-stimulated production of nitric oxide in endothelial cells.
AuthorsMontagnani M, Ravichandran LV, Chen H, Esposito DL, Quon MJ
JournalMol Endocrinol
PubMed ID12145346
Vasodilator actions of insulin are mediated by signaling pathways involving phosphatidylinositol 3-kinase (PI 3-kinase) and Akt that lead to activation of endothelial nitric oxide synthase (eNOS) in endothelium. Signaling molecules immediately upstream and downstream from PI 3-kinase involved with production of NO in response to insulin have not been previously ... More
Nitric oxide mediated photo-induced cell death in human malignant cells.
AuthorsAli SM, Olivo M
JournalInt J Oncol
PubMed ID12632064
Photodynamic therapy (PDT) is a therapeutic modality used for the treatment of a variety of solid neoplasms. The principle of PDT is based on the selective uptake of a photosensitizing chemical in tumor tissue/cell followed by irradiation of tumors with visible light. The treatment results in a cascade of oxidative ... More
Oxyresveratrol and resveratrol are potent antioxidants and free radical scavengers: effect on nitrosative and oxidative stress derived from microglial cells.
AuthorsLorenz P, Roychowdhury S, Engelmann M, Wolf G, Horn TF
JournalNitric Oxide
PubMed ID14623172
Hydroxystilbenes are naturally occurring polyphenols with protective effects against reactive oxygen and nitrogen species (ROS/RNS). Here, we investigated oxyresveratrol (OXY), which is contained in high amounts in mulberry wood, in comparison to the antioxidant resveratrol (RES). We found that OXY is a more effective scavenger for 2,2-diphenyl-1-picryl-hydrazyl (DPPH, 100 microM) ... More
Platelet-activating factor synthesis by neutrophils, monocytes, and endothelial cells is modulated by nitric oxide production.
AuthorsMariano F, Bussolati B, Migliori M, Russo S, Triolo G, Camussi G
JournalShock
PubMed ID12688545
Nitric oxide (NO) and platelet-activating factor (PAF) can modulate the interaction between endothelial lining and circulating leukocytes. Several studies implicated the production of PAF and NO in the pathogenesis of microcirculatory alterations occurring in septic shock. However, the reciprocal interaction between PAF and NO has not been fully elucidated. In ... More
Nitric oxide (NO) synthase mRNA expression and NO production via muscarinic acetylcholine receptor-mediated pathways in the CEM, human leukemic T-cell line.
AuthorsKamimura Y, Fujii T, Kojima H, Nagano T, Kawashima K
JournalLife Sci
PubMed ID12628472
Nitric oxide (NO) is synthesized from L-arginine by neuronal, endothelial and inducible isoforms of NO synthase (nNOS, eNOS and iNOS, respectively) and is involved in the regulation of a variety of physiological functions, including immune activity. In vascular endothelial cells, stimulation of M(3) subtype of muscarinic acetylcholine receptors (mAChRs) triggers ... More
High-salt diet depresses acetylcholine reactivity proximal to NOS activation in cerebral arteries.
Rats were fed a low-salt (LS; 0.4% NaCl) or high-salt (HS; 4.0% NaCl) diet for 3 days, and the responses of isolated cerebral arteries to acetylcholine (ACh), the nitric oxide (NO)-dependent dilator bradykinin, and the NO donor 6-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-hex-anamine (NOC-9) were determined. ACh-induced vasodilation and NO release, assessed with the fluorescent ... More
Increased nitric oxide production by neutrophils from patients with chronic granulomatous disease on trimethoprim-sulfamethoxazole.
AuthorsTsuji S, Taniuchi S, Hasui M, Yamamoto A, Kobayashi Y
JournalNitric Oxide
PubMed ID12446177
Chronic granulomatous disease (CGD) is an inherited disease characterized by severe and recurrent bacterial and fungal infections. Phagocytic cells of CGD patients are unable to produce superoxide anion, and their efficiency in bacterial killing is significantly impaired. In these patients, the prophylactic and therapeutic validity of a long-term use of ... More
Hyperglycemic switch from mitochondrial nitric oxide to superoxide production in endothelial cells.
AuthorsBrodsky SV, Gao S, Li H, Goligorsky MS
JournalAm J Physiol Heart Circ Physiol
PubMed ID12384491
The accumulated ultrastructural and biochemical evidence is highly suggestive of the existence of mitochondrial nitric oxide (NO) synthase (mtNOS), where local production of NO regulates the electron transport along the respiratory chain. Here, the functional competence of mtNOS in situ in a living cell was examined using an intravital fluorescent ... More
Effect of SNI-2011 on amylase secretion from parotid tissue in rats and in neuronal nitric oxide synthase knockout mice.
AuthorsYuan Z, Iida H, Inoue N, Ishikawa Y, Ishida H
JournalEur J Pharmacol
PubMed ID12620514
The effect of (+/-)cis-2-methylspilo(1,3-oxathiolane-5,3')quinuclidine (SNI-2011) on the secretory pathway of amylase in parotid tissues was investigated. SNI-2011-induced exocytosis was inhibited by a cell-permeable Ca(2+) chelator or inhibitors of calmodulin kinase II, neuronal nitric oxide synthase (nNOS), soluble guanyl cyclase, cyclic GMP-dependent protein kinase (PKG), and myosin light chain kinase, suggesting ... More
Orthogonality of calcium concentration and ability of 4,5-diaminofluorescein to detect NO.
AuthorsSuzuki N, Kojima H, Urano Y, Kikuchi K, Hirata Y, Nagano T
JournalJ Biol Chem
PubMed ID11641405
We have developed diaminofluoresceins (DAFs) and diaminorhodamines as fluorescent indicators for NO based on the specific reaction of the aromatic vicinal diamines with NO. Among them, 4,5-diaminofluorescein (DAF-2) is widely used for real-time biological imaging of NO in cultured cells or tissues by many researchers. Contrary to a recent report ... More
Visualization of nitric oxide production in the earthworm ventral nerve cord.
AuthorsKitamura Y, Naganoma Y, Horita H, Tsuji N, Shimizu R, Ogawa H, Oka K
JournalNeurosci Res
PubMed ID11377756
Distribution of nitric oxide (NO)-producible neurons in the ventral nerve cord (VNC) of the earthworm was investigated by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry. Some neurons (20-30 microm in diameter) were intensely stained and were localized in areas between the 1st and 2nd lateral nerves in the ventral side ... More
Transcriptional stimulation of the eNOS gene by the stable prostacyclin analogue beraprost is mediated through cAMP-responsive element in vascular endothelial cells: close link between PGI2 signal and NO pathways.
AuthorsNiwano K, Arai M, Tomaru K, Uchiyama T, Ohyama Y, Kurabayashi M
JournalCirc Res
PubMed ID12919953
Beraprost sodium (BPS), an orally active prostacyclin analogue, has been reported to be beneficial in the treatment of primary pulmonary hypertension and obstructive peripheral arterial disease. Although BPS was originally described for its effects on platelet aggregation and vasodilatory response, the effect on endothelial cells has been poorly understood. In ... More
Inhibition by (+/-)-indenestrol A of interferon gamma-stimulated nitric oxide formation in murine macrophage RAW 264.7 cells.
AuthorsOda T, So Y, Sato Y, Shimizu N, Handa H, Yasukochi Y, Kasahara T
JournalMutat Res
PubMed ID12504767
We examined the effects of (+/-)-indenestrol A (IA), an antioxidative and superoxide-producing metabolite of diethylstilbestrol (DES), on the activation of murine macrophages (RAW 264.7 cells) in vitro, particularly with regard to interferon (IFN)-gamma-induced nitric oxide (NO) production. (+/-)-IA inhibited NO production more strongly than DES as assessed by a nitrite ... More
Insulin-stimulated activation of eNOS is independent of Ca2+ but requires phosphorylation by Akt at Ser(1179).
AuthorsMontagnani M, Chen H, Barr VA, Quon MJ
JournalJ Biol Chem
PubMed ID11402048
Vasodilator actions of insulin are mediated by activation of endothelial nitric-oxide synthase (eNOS) and subsequent production of NO. Phosphatidylinositol 3-kinase and Akt play important roles in insulin-signaling pathways leading to production of NO in vascular endothelium. Here we dissected mechanisms whereby insulin activates eNOS by using the fluorescent dye DAF-2 ... More
High-salt diet increases sensitivity to NO and eNOS expression but not NO production in THALs.
L-Arginine inhibits thick ascending limb (THAL) NaCl absorption by activating endothelial NO synthase (eNOS) and increasing NO production. Inhibition of renal NO production combined with a high-salt diet produces hypertension, and the THAL has been implicated in salt-sensitive hypertension. We hypothesized that a high-salt diet enhances the inhibitory action of ... More
Nitrosative stress by cyclosporin A in the endothelium: studies with the NO-sensitive probe diaminofluorescein-2/diacetate using flow cytometry.
AuthorsNavarro-Antolin J, Lamas S
JournalNephrol Dial Transplant
PubMed ID11369812
BACKGROUND: We have reported previously that exposure of endothelial cells to cyclosporin A (CsA) may result in the regulation of specific genes, such as the endothelial nitric oxide synthase. In the context of endothelial toxicity, whether this represents an adaptive response to injury at the physiological level or contributes further ... More
Nicotinic receptor alpha 7-subunits are coupled to the stimulation of nitric oxide synthase in rat dorsal root ganglion neurons.
AuthorsHaberberger RV, Henrich M, Lips KS, Kummer W
JournalHistochem Cell Biol
PubMed ID12898272
In dorsal root ganglia (DRG) intraganglionic communication takes place both among neurons and between neurons and satellite cells. One diffusible substance involved in this signalling is nitric oxide (NO), and acetylcholine (ACh) is a candidate for the stimulation of intraganglionic NO synthesis. DRG neurons react to ACh-receptor stimulation with NO-dependent ... More
Involvement of hydrogen peroxide and nitric oxide in expression of the ipomoelin gene from sweet potato.
AuthorsJih PJ, Chen YC, Jeng ST
JournalPlant Physiol
PubMed ID12746543
The IPO (ipomoelin) gene was isolated from sweet potato (Ipomoea batatas cv Tainung 57) and used as a molecular probe to investigate its regulation by hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) after sweet potato was wounded. The expression of the IPO gene was stimulated by H(2)O(2) whether or not ... More
Determinants of shear stress-stimulated endothelial nitric oxide production assessed in real-time by 4,5-diaminofluorescein fluorescence.
AuthorsQiu W, Kass DA, Hu Q, Ziegelstein RC
JournalBiochem Biophys Res Commun
PubMed ID11500041
The extremely short biological half-life of endothelial-derived nitric oxide (NO) has impeded real-time measurements of NO synthesis. We used the membrane-permeable fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA) to study determinants of NO synthesis in bovine aortic endothelial cells (BAECs). A step increase in shear stress (SS) from 0.3 to 3.4 ... More
Mislocalization of eNOS and upregulation of cerebral vascular Ca2+ channel activity in angiotensin-hypertension.
AuthorsGerzanich V, Ivanova S, Zhou H, Simard JM
JournalHypertension
PubMed ID12668586
We tested the hypothesis that endothelial dysfunction induced by angiotensin II (Ang-hypertension) would impair regulatory control of vascular smooth muscle L-type Ca2+ channels by endothelial nitric oxide synthase (eNOS). We studied cerebral lenticulostriate arterioles (LSAs) from control rats, from rats infused with Ang (240 microg x kg(-1) x h(-1) SQ ... More
Reliable in vitro measurement of nitric oxide released from endothelial cells using low concentrations of the fluorescent probe 4,5-diaminofluorescein.
AuthorsLeikert JF, Räthel TR, Müller C, Vollmar AM, Dirsch VM
JournalFEBS Lett
PubMed ID11591386
4,5-Diaminofluorescein (DAF-2) and its membrane-permeable derivate DAF-2 diacetate are fluorescent probes that have been developed to perform real-time biological detection of nitric oxide (NO). Their use for intracellular imaging, however, has recently been seriously questioned and data using DAF-2 for extracellular NO detection at low levels, as for example released ... More
Interfering with nitric oxide measurements. 4,5-diaminofluorescein reacts with dehydroascorbic acid and ascorbic acid.
AuthorsZhang X, Kim WS, Hatcher N, Potgieter K, Moroz LL, Gillette R, Sweedler JV
JournalJ Biol Chem
PubMed ID12370177
4,5-Diaminofluorescein (DAF-2) is widely used for detection and imaging of NO based on its sensitivity, noncytotoxicity, and specificity. In the presence of oxygen, NO and NO-related reactive nitrogen species nitrosate 4,5-diaminofluorescein to yield the highly fluorescent DAF-2 triazole (DAF-2T). However, as reported here, the DAF-2 reaction to form a fluorescent ... More
Nonendothelial source of nitric oxide in arterioles but not in venules: alternative source revealed in vivo by diaminofluorescein microfluorography.
AuthorsKashiwagi S, Kajimura M, Yoshimura Y, Suematsu M
JournalCirc Res
PubMed ID12480826
This study aimed to examine topographic distribution of microvascular NO generation in vivo. To this end, nitrosonium ion (NO+)-sensitive diaminofluorescein diacetate was superfused continuously on the rat mesentery and the fluorescence was visualized in the microvessels through laser confocal microfluorography. Two major sites exhibited a time-dependent elevation of the fluorescence: ... More
Role of ascorbate in the regulation of nitric oxide generation by polymorphonuclear leukocytes.
AuthorsSharma P, Raghavan SA, Dikshit M
JournalBiochem Biophys Res Commun
PubMed ID12943656
We have recently demonstrated that NO-mediated polymorphonuclear (PMN)-dependent inhibition of rat platelet aggregation is significantly enhanced in the presence of ascorbate. Consequently, the present study was undertaken to elucidate the underlying mechanisms involved in ascorbate-mediated potentiation of NO synthesis in PMNs. We observed that ascorbate or its oxidized product, dehydroascorbate ... More
A fluorescence-based method for measuring nitric oxide in extracts of skeletal muscle.
AuthorsSutherland H, Khundkar R, Zolle O, McArdle A, Simpson AW, Jarvis JC, Salmons S
JournalNitric Oxide
PubMed ID11587562
We describe here a fluorescence assay for nitric oxide synthase activity in skeletal muscle based on a new indicator, 4,5-diaminofluorescein (DAF-2). The rapid and irreversible binding of DAF-2 to oxidized NO allows real-time measurement of NO production. The method is safer and more convenient than the usual citrulline radioassay and ... More
Urotensin II is a nitric oxide-dependent vasodilator and natriuretic peptide in the rat kidney.
AuthorsZhang AY, Chen YF, Zhang DX, Yi FX, Qi J, Andrade-Gordon P, de Garavilla L, Li PL, Zou AP
JournalAm J Physiol Renal Physiol
PubMed ID12783779
Recent studies have indicated that urotensin II (UII), a cyclic peptide, is vasoactive and may be involved in cardiovascular dysfunctions. It remains unknown, however, whether UII plays a role in the control of renal vascular tone and tubular function. In the present study, a continuous infusion of synthetic human UII ... More
Arrest of B16 melanoma cells in the mouse pulmonary microcirculation induces endothelial nitric oxide synthase-dependent nitric oxide release that is cytotoxic to the tumor cells.
AuthorsQiu H, Orr FW, Jensen D, Wang HH, McIntosh AR, Hasinoff BB, Nance DM, Pylypas S, Qi K, Song C, Muschel RJ, Al-Mehdi AB
JournalAm J Pathol
PubMed ID12547699
Metastatic cancer cells seed the lung via blood vessels. Because endothelial cells generate nitric oxide (NO) in response to shear stress, we postulated that the arrest of cancer cells in the pulmonary microcirculation causes the release of NO in the lung. After intravenous injection of B16F1 melanoma cells, pulmonary NO ... More
Oxidized LDL inhibits vascular endothelial growth factor-induced endothelial cell migration by an inhibitory effect on the Akt/endothelial nitric oxide synthase pathway.
AuthorsChavakis E, Dernbach E, Hermann C, Mondorf UF, Zeiher AM, Dimmeler S
JournalCirculation
PubMed ID11319202
BACKGROUND: Oxidized LDL (oxLDL) inhibits endothelial cell (EC) migration. Stimulating ECs with vascular endothelial growth factor (VEGF) leads to the activation of Akt/protein kinase B, which in turn activates endothelial nitric oxide synthase (eNOS) by phosphorylation on serine 1177. VEGF-induced cell migration is dependent on the generation of nitric oxide ... More
Bioimaging of nitric oxide with fluorescent indicators based on the rhodamine chromophore.
AuthorsKojima H, Hirotani M, Nakatsubo N, Kikuchi K, Urano Y, Higuchi T, Hirata Y, Nagano T
JournalAnal Chem
PubMed ID11354477
Diaminofluoresceins are widely used for detection and imaging of nitric oxide (NO), but for biological applications, they have the disadvantages that the fluorescence of the fluorescein chromophore is pH-sensitive and overlaps the autofluorescence of cells. We have developed a membrane-permeable fluorescent indicator for NO based on the rhodamine chromophore, DAR-4M ... More
Attenuation of neuropathic pain by the nociceptin/orphanin FQ antagonist JTC-801 is mediated by inhibition of nitric oxide production.
AuthorsMabuchi T, Matsumura S, Okuda-Ashitaka E, Kitano T, Kojima H, Nagano T, Minami T, Ito S
JournalEur J Neurosci
PubMed ID12713641
At the spinal level, the involvement of nociceptin/orphanin FQ (N/OFQ) in pain transmission is controversial. JTC-801, a selective nonpeptidergic N/OFQ antagonist, is a good tool to examine the involvement of endogenous N/OFQ in pathophysiological conditions. In the present study, we studied the effect of JTC-801 on neuropathic pain induced by ... More
Cooperation between insulin and leptin in the modulation of vascular tone.
AuthorsVecchione C, Aretini A, Maffei A, Marino G, Selvetella G, Poulet R, Trimarco V, Frati G, Lembo G
JournalHypertension
PubMed ID12835332
High levels of insulin and leptin have been reported in human hypertension, suggesting a role for these metabolic hormones in blood pressure homeostasis. These hormones interact on intermediate metabolism, but nothing is known about their interaction at the vascular level. Our data demonstrate that insulin (0.6 nmol/L) is able to ... More
Mitochondrial nitric oxide localization in H9c2 cells revealed by confocal microscopy.
AuthorsZanella B, Calonghi N, Pagnotta E, Masotti L, Guarnieri C
JournalBiochem Biophys Res Commun
PubMed ID11798175
This study shows the presence of all three nitric oxide synthases (NOSs) and NOS activity in H9c2 cells cultured under non-stimulated conditions. By using the 4,5 diaminofluoresceindiacetate (DAF-2DA) fluorimetric nitric oxide (NO(*)) detection system we observed NO(*) production in H9c2 cells. As revealed by confocal microscopy, NO(*) fluorescence colocalizes in ... More
ROS and NO trigger early preconditioning: relationship to mitochondrial KATP channel.
Reactive oxygen species (ROS) and nitric oxide (NO) are implicated in induction of ischemic preconditioning. However, the relationship between these oxidant signals and opening of the mitochondrial ATP-dependent potassium (K(ATP)) channel during early preconditioning is not fully understood. We observed preconditioning protection by hypoxia, exogenous H(2)O(2), or PKC activator PMA ... More
Characterization of basal nitric oxide production in living cells.
AuthorsLópez-Figueroa MO, Caamaño C, Marin R, Guerra B, Alonso R, Morano MI, Akil H, Watson SJ
JournalBiochim Biophys Acta
PubMed ID11583820
Nitric oxide (NO) is an important modulator of immune, endocrine and neuronal functions; however, measuring physiological levels of NO in cell cultures is generally difficult because of the lack of suitable methodologies. We have selected three cell lines from different origins: the neuroblastoma-derived Neuro2A (N2A), the cholinergic SN56 and the ... More
Nitric oxide production within rat urothelial cells.
AuthorsMastrangelo D, Baertschi AJ, Roatti A, Amherdt M, Iselin CE
JournalJ Urol
PubMed ID14501779
PURPOSE: Recent studies have suggested that nitric oxide (NO) synthase (NOS) may be localized in the urothelium of the proximal part of the mammalian ureter. We investigated endogenous NO production in the proximal half of the rat ureter, localized its cellular source, characterized the NOS isoforms involved and assessed the ... More
Tumor cell apoptosis by irradiation-induced nitric oxide production in vascular endothelium.
AuthorsHirakawa M, Oike M, Masuda K, Ito Y
JournalCancer Res
PubMed ID11888919
We examined the effects of X-ray irradiation on endothelial nitric oxide (NO) production in bovine aortic endothelial cells (BAECs). Single irradiation of up to 60 Gy did not affect the expression of endothelial NO synthase (eNOS) mRNA, as assessed by reverse transcription-PCR, in BAECs. The basal level of intracellular Ca(2+) ... More
Nitric oxide and reactive oxygen species do not elicit hypersensitive cell death but induce apoptosis in the adjacent cells during the defense response of oat.
AuthorsTada Y, Mori T, Shinogi T, Yao N, Takahashi S, Betsuyaku S, Sakamoto M, Park P, Nakayashiki H, Tosa Y, Mayama S
JournalMol Plant Microbe Interact
PubMed ID15000391
Nitric oxide (NO) acts as a signaling molecule in many cellular responses in plants and animals. Oat plants (Avena sativa L.) evoke the hypersensitive response (HR), which shares morphological and biochemical features with mammalian apoptosis, such as DNA laddering and heterochromatin condensation, in response to the avirulent crown rust fungus ... More
Flavopiridol downregulates the expression of both the inducible NO synthase and p27(kip1) in malignant cells from B-cell chronic lymphocytic leukemia.
Flavopiridol, an inhibitor of cyclin-dependent kinases and other protein kinases, induces in vitro apoptosis of malignant cells from B-cell chronic lymphocytic leukemia (B-CLL). Previously, we reported that nitric oxide (NO), produced by an inducible NO synthase (iNOS), spontaneously expressed by the B-CLL cells, contributed to their deficiency in apoptosis. In ... More
Nitric oxide is induced by wounding and influences jasmonic acid signaling in Arabidopsis thaliana.
Nitric oxide (NO) has been associated with plant defense responses during microbial attack, and with induction and/or regulation of programmed cell death. Here, we addressed whether NO participates in wound responses in Arabidopsis thaliana (L.) Heynh. Real-time imaging by confocal laser-scanning microscopy in conjunction with the NO-selective fluorescence indicator 4,5-diaminofluorescein ... More
Distinct glutamate receptors govern differential levels of nitric oxide production in a layer-specific manner in the rat cerebellar cortex.
AuthorsOkada D, Yap CC, Kojima H, Kikuchi K, Nagano T
JournalNeuroscience
PubMed ID15062988
To evaluate roles of nitric oxide (NO) in neural functions, it is critical to know how neural inputs activate neuronal NO synthase in individual sites. Although NMDA receptor-dependent mechanism well explains postsynaptic, robust NO production, this sole mechanism does not explain some aspects of NO production in the brain, such ... More
Constitutive nitric oxide production in bovine aortic and brain microvascular endothelial cells: a comparative study.
AuthorsKimura C, Oike M, Ohnaka K, Nose Y, Ito Y
JournalJ Physiol
PubMed ID14617679
Vascular endothelium constitutively generates nitric oxide (NO) in large vessels and induces a relaxation of smooth muscle cells. However, little is known about the production of NO in microvessels, where smooth muscle layers are thin or absent. In this study, we have compared the constitutive production of NO in bovine ... More
Application of 4,5-diaminofluorescein to reliably measure nitric oxide released from endothelial cells in vitro.
AuthorsRäthel TR, Leikert J Vollmar AM, Dirsch VM
JournalBiol Proced Online
PubMed ID14569611
Here we describe in more depth the previously published application of the fluorescent probe 4,5-diaminofluorescein (DAF-2) in order to reliably measure low levels of nitric oxide (NO) as released from human endothelial cells in vitro. The used approach is based on the following considerations a) use low concentrations of DAF-2 ... More
High concentration of glucose inhibits glomerular endothelial eNOS through a PKC mechanism.
AuthorsChu S, Bohlen HG
JournalAm J Physiol Renal Physiol
PubMed ID15140758
Kidney glomeruli are important targets of diabetic nephropathy. We hypothesized a high concentration of glucose could suppress glomerular endothelial nitric oxide synthase (eNOS) by a protein kinase C (PKC) mechanism, as has been found in other tissues. Mouse kidney slices (150-200 microm) were bathed in Hanks' solution with 100 microM ... More
Luminal flow induces eNOS activation and translocation in the rat thick ascending limb. II. Role of PI3-kinase and Hsp90.
AuthorsOrtiz PA, Hong NJ, Garvin JL
JournalAm J Physiol Renal Physiol
PubMed ID15100099
Endothelial nitric oxide synthase (eNOS) regulates NaCl absorption by the thick ascending limb of the loop of Henle (THAL). We found that augmenting luminal flow induces eNOS activation and translocation to the apical membrane of THALs (Ortiz PA, Hong NJ, and Garvin JL. Am J Physiol Renal Physiol 287: F274-F280, ... More
Impaired nitric oxide production of the myenteric plexus in colitis detected by a new bioimaging system.
AuthorsKono T, Chisato N, Ebisawa Y, Asama T, Sugawara M, Ayabe T, Kohgo Y, Kasai S, Yoneda M, Takahashi T
JournalJ Surg Res
PubMed ID15047139
Direct measurement of the release of nitric oxide (NO) from the myenteric plexus has been extremely difficult to date, due to the lack of suitable methodologies. We have developed a new bioimaging system to visualize the nitrergic neurons of the myenteric plexus and investigated whether NO production is impaired in ... More
Nitric oxide imaging in neurons using confocal microscopy.
AuthorsZheng X, Ning G, Hong D, Zhang M
JournalMethods Mol Biol
PubMed ID15199237
The instability and low concentration of nitric oxide (NO) in specimens make it difficult to be detected directly. In this chapter, a method for imaging nitric oxide using laser scanning confocal microscopy (LSCM) is presented. A cultured hippocampal neuron is dyed with DAF-2 and observed under a Zeiss LSM 510 ... More
Proinsulin C-peptide increases nitric oxide production by enhancing mitogen-activated protein-kinase-dependent transcription of endothelial nitric oxide synthase in aortic endothelial cells of Wistar rats.
AuthorsKitamura T, Kimura K, Makondo K, Furuya DT, Suzuki M, Yoshida T, Saito M
JournalDiabetologia
PubMed ID14586499
AIMS/HYPOTHESIS: Recent studies have suggested that proinsulin C-peptide improves vascular functions, possibly through nitric oxide (NO) production. To clarify the molecular mechanisms of vascular NO production induced by C-peptide, we examined the effects of C-peptide on NO production and NO synthase expression in rat aortic endothelial cells in connection with ... More
Constitutive nitric oxide synthases in rat pancreatic islets: direct imaging of glucose-induced nitric oxide production in beta-cells.
AuthorsNakada S, Ishikawa T, Yamamoto Y, Kaneko Y, Nakayama K
JournalPflugers Arch
PubMed ID14564523
The production of nitric oxide (NO) by constitutive nitric oxide synthase (NOS) was investigated in isolated rat pancreatic islets and dispersed beta-cells. Double-immunocytochemical analyses with a confocal microscope demonstrated the presence of NOS1 in alpha-, beta-, delta-, and PP-cells and that of NOS3 in beta-, delta-, and PP-cells, but not ... More
Nicotinic receptor mediated stimulation of NO-generation in neurons of rat thoracic dorsal root ganglia.
The contribution of nicotinic acetylcholine receptors (nAChRs) to stimulation of NO-production was investigated in isolated rat dorsal root ganglion (DRG) neurons utilizing an NO-sensitive fluorescent indicator 4,5-diaminofluorescein-diacetate (DAF-2DA) and appropriate channel blockers. RT-PCR and immunohistochemical analysis of NOS isoforms in cultured neurons revealed the expression of eNOS in the vast ... More
C. elegans SGK-1 is the critical component in the Akt/PKB kinase complex to control stress response and life span.
AuthorsHertweck M, Göbel C, Baumeister R
JournalDev Cell
PubMed ID15068796
The DAF-2 insulin receptor-like signaling pathway controls metabolism, development, longevity, and stress response in C. elegans. Here we show that SGK-1, the C. elegans homolog of the serum- and glucocorticoid-inducible kinase SGK, acts in parallel to the AKT kinases to mediate DAF-2 signaling. Loss of sgk-1 results in defective egg-laying, ... More
Signaling mechanisms that mediate nitric oxide production induced by acetylcholine exposure and withdrawal in cat atrial myocytes.
AuthorsDedkova EN, Ji X, Wang YG, Blatter LA, Lipsius SL
JournalCirc Res
PubMed ID14615286
Fluorescence microscopy and the NO-sensitive indicator 4,5-diaminofluorescein were used to determine the effects of acetylcholine (ACh) on intracellular NO (NOi) in cat atrial myocytes. Field stimulation (1 Hz) of cells or exposure of quiescent cells to ACh (1 to 10 micromol/L) had no effect on NOi. However, in field-stimulated cells, ... More
Acute and chronic regulation of thick ascending limb endothelial nitric oxide synthase by statins.
AuthorsVarela M, Garvin JL
JournalJ Am Soc Nephrol
PubMed ID14747373
Statins, 3-hydroxy-3-methylglutaryl CoA reductase inhibitors, acutely increase endothelial nitric oxide synthase (eNOS) activity and chronically increase eNOS expression in endothelial cells. NO decreases transport in thick ascending limbs (TAL). We hypothesized that statins inhibit TAL transport by acutely activating eNOS, thereby increasing NO production and chronically enhancing eNOS expression. Oxygen ... More
A nonradioactive assay for nitric oxide synthase activity in tissue extracts.
AuthorsSutherland H, Zolle O, Khundkar R, Simpson AW, Jarvis JC, Salmons S
JournalMethods Mol Biol
PubMed ID15199239
We describe a fluorescence assay for nitric oxide synthase activity based on a new indicator, 4,5-diaminofluorescein (DAF-2). The method offers the advantage of being safer and more convenient than the citrulline radioassay in common use. The rapid and irreversible binding of DAF-2 to oxidized nitric oxide (NO) enables NO production ... More
Direct evidence that induced nitric oxide production in hepatocytes prevents liver damage during lipopolysaccharide tolerance in rats.
AuthorsEbisawa Y, Kono T, Yoneda M, Asama T, Chisato N, Sugawara M, Ishikawa K, Iwamoto J, Ayabe T, Kohgo Y, Kasai S
JournalJ Surg Res
PubMed ID15100007
BACKGROUND: The role of nitric oxide (NO) in lipopolysaccharide (LPS) tolerance in the liver has been investigated in a number of previous studies, but it is still not clear whether NO is cytotoxic or cytoprotective. The aims of this study were to investigate whether low-dose LPS (LLPS)-induced hepatic production of ... More
Nitric oxide induces cell death in Taxus cells.
AuthorsPedroso MC, Magalhaes JR, Durzan D
JournalPlant Sci
PubMed ID10960730
Sodium nitroprusside (SNP), a nitric oxide donor, or centrifugation at 150 times unit gravity, caused a nitric oxide burst in oocyte-derived Taxus brevifolia haploid cultures. This burst, visualized by the specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA), preceded a significant increase in nuclear DNA fragmentation and cell death. DNA fragmentation ... More
Effect of high-salt diet on NO release and superoxide production in rat aorta.
AuthorsZhu J, Mori T, Huang T, Lombard JH
JournalAm J Physiol Heart Circ Physiol
PubMed ID14527935
Sprague-Dawley rats were fed either a high-salt (HS) diet (4.0% NaCl) or a low-salt (LS) diet (0.4% NaCl) for 3 days. Nitric oxide (NO) and superoxide production were assessed in the thoracic aorta by evaluating the fluorescence signal intensity from 4,5-diaminofluorescein (DAF-2DA) and dihydroethidine, respectively. Methacholine caused increased NO release ... More
Altered production of nitric oxide and reactive oxygen species in rat nodose ganglion neurons during acute hypoxia.
AuthorsYamamoto Y, Henrich M, Snipes RL, Kummer W
JournalBrain Res
PubMed ID12535770
'Nitric oxide (NO) production in the sensory neurons of the rad nodose ganglion was studied by examining the distribuiotn of NO synthase (NOS) by use of NADPH diaphorase (NADPHD) histochemistry and immunohistochemistry ofr the presence of isoformes of NOS: neuronal (nNOS), endothelial (eNOS) and the inducible isoform (iNOS). Distribution and ... More
A stress surveillance system based on calcium and nitric oxide in marine diatoms.
AuthorsVardi A, Formiggini F, Casotti R, De Martino A, Ribalet F, Miralto A, Bowler C
JournalPLoS Biol
PubMed ID16475869
'Diatoms are an important group of eukaryotic phytoplankton, responsible for about 20% of global primary productivity. Study of the functional role of chemical signaling within phytoplankton assemblages is still in its infancy although recent reports in diatoms suggest the existence of chemical-based defense strategies. Here, we demonstrate how the accurate ... More
Focusing of nitric oxide mediated nitrosation and oxidative nitrosylation as a consequence of reaction with superoxide.
AuthorsEspey MG, Thomas DD, Miranda KM, Wink DA
JournalProc Natl Acad Sci U S A
PubMed ID12177414
'The impact of nitric oxide (NO) synthesis on different biological cascades can rapidly change dependent on the rate of NO formation and composition of the surrounding milieu. With this perspective, we used diaminonaphthalene (DAN) and diaminofluorescein (DAF) to examine the nitrosative chemistry derived from NO and superoxide (O2-) simultaneously generated ... More
Nitric oxide-dependent mitochondrial biogenesis generates Ca2+ signaling profile of lupus T cells.
AuthorsNagy G, Barcza M, Gonchoroff N, Phillips PE, Perl A
JournalJ Immunol
PubMed ID15356113
'Abnormal T cell activation and cell death underlie the pathology of systemic lupus erythematosus. Although mitochondrial hyperpolarization (MHP) represents an early and reversible checkpoint of T cell activation and apoptosis, lupus T cells exhibit persistent MHP. NO has recently been recognized as a key signal of mitochondrial biogenesis and mediator ... More
Fluorescence monitoring of ATP-stimulated, endothelium-derived nitric oxide production in channels of a poly(dimethylsiloxane)-based microfluidic device.
AuthorsD'Amico Oblak T, Root P, Spence DM
JournalAnal Chem
PubMed ID16643013
'Intracellular nitric oxide (NO) production in a microfluidic endothelium is detected using fluorescence microscopy. Bovine pulmonary artery endothelial cells (bPAECs) were loaded with the fluorescence probe diaminodifluorofluorescein diacetate (DAF-FM DA), and the subsequent fluorescent DAF-FM DA/NO adduct was measured. Solutions of bradykinin, a well-known stimulus of endothelium-derived NO, activated nitric ... More
Mitochondrial nitric oxide synthase is not eNOS, nNOS or iNOS.
AuthorsLacza Z, Snipes JA, Zhang J, Horváth EM, Figueroa JP, Szabó C, Busija DW
JournalFree Radic Biol Med
PubMed ID14607521
'Recent studies indicated that there is a distinct mitochondrial nitric oxide synthase (mtNOS) enzyme, which may be identical to the other known NOS isoforms. We investigated the possible involvement of the endothelial, the neuronal, and the inducible NOS isoforms (eNOS, nNOS, iNOS, respectively) in mitochondrial NO production. Mouse liver mitochondria ... More
In vivo imaging of an elicitor-induced nitric oxide burst in tobacco.
AuthorsFoissner I, Wendehenne D, Langebartels C, Durner J
JournalPlant J
PubMed ID10998192
'A growing body of evidence suggests that nitric oxide (NO), an important signalling and defence molecule in mammals, plays a key role in activating disease resistance in plants, acting as signalling molecule and possibly as direct anti-microbial agent. Recently, a novel fluorophore (diaminofluorescein diacetate, DAF-2 DA) has been developed which ... More
Real-time measurement of nitric oxide in single mature mouse skeletal muscle fibres during contractions.
AuthorsPye D, Palomero J, Kabayo T, Jackson MJ
JournalJ Physiol
PubMed ID17331997
'Nitric oxide (NO) is thought to play multiple roles in skeletal muscle including regulation of some adaptations to contractile activity, but appropriate methods for the analysis of intracellular NO activity are lacking. In this study we have examined the intracellular generation of NO in isolated single mature mouse skeletal muscle ... More
Kinetic characterization of the nitric oxide toxicity for PC12 cells: effect of half-life time of NO release.
AuthorsYamamoto T, Yuyama K, Nakamura K, Kato T, Yamamoto H
JournalEur J Pharmacol
PubMed ID10844095
'We investigated the effects of low concentrations of nitric oxide (NO) on cell viability using NO donors, (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hex enamid e (NOR1), (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR2), (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR3) and (+/-)-N-[(E)-4-ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexen-1- yl]-3-pyr idine (NOR4). The half-life times of the NO release from these four NOR analogs, NOR1, NOR2, NOR3 and NOR4, were determined (6.5, ... More
Examining intracellular organelle function using fluorescent probes: from animalcules to quantum dots.
AuthorsZorov DB, Kobrinsky E, Juhaszova M, Sollott SJ
JournalCirc Res
PubMed ID15297386
'Fluorescence microscopy imaging has become one of the most useful techniques to assess the activity of individual cells, subcellular trafficking of signals to and between organelles, and to appreciate how organelle function is regulated. The past 2 decades have seen a tremendous advance in the rational design and development in ... More
Elevated glucose concentrations promote receptor-independent activation of adherent human neutrophils: an experimental and computational approach.
AuthorsKummer U, Zobeley J, Brasen JC, Fahmy R, Kindzelskii AL, Petty AR, Clark AJ, Petty HR
JournalBiophys J
PubMed ID17237194
'Neutrophil activation plays integral roles in host tissue damage and resistance to infectious diseases. As glucose uptake and NADPH availability are required for reactive oxygen metabolite production by neutrophils, we tested the hypothesis that pathological glucose levels (>or=12 mM) are sufficient to activate metabolism and reactive oxygen metabolite production in ... More
'The source of superoxide (O2*-) production and cell-to-cell interactions of O2*- and nitric oxide (NO) in response to angiotensin II (AngII) were studied by fluorescence microscopic techniques to image rat renal outer medullary microtissue strips. Changes in intracellular O2*- were determined by dihydroethidium-ethidium ratios, and NO was determined with 4,5-diaminofluorescein ... More
Fluorescent Indicators for Imaging Nitric Oxide Production.
AuthorsKojima H, Urano Y, Kikuchi K, Higuchi T, Hirata Y, Nagano T
JournalAngew Chem Int Ed Engl
PubMed ID10556905
'The membrane-permeating indicator DAF-FM DA is transformed by intracellular esterases into the highly water-soluble dye DAF-FM, which traps NO produced by NO synthase (NOS) to yield a highly fluorescent triazole compound in cells (see schematic diagram). Monitoring with a fluorescence microscope should allow direct identification of intracellular production and location ... More
Nitric oxide production in living neurons is modulated by sphingosine: a fluorescence microscopy study.
AuthorsViani P, Giussani P, Ferraretto A, Signorile A, Riboni L, Tettamanti G
JournalFEBS Lett
PubMed ID11602242
'An investigation was carried out into the possible effect of sphingosine (Sph) on nitric oxide (NO) production in living neurons. Differentiated granule cells were used in a dynamic videoimaging analysis of single cells labeled, simultaneously, with FURA-2 and the NO indicator 4,5-diaminofluorescein. The results demonstrate that Sph exerts a potent ... More
What is the real physiological NO concentration in vivo?
AuthorsHall CN, Garthwaite J,
JournalNitric Oxide
PubMed ID19602444
'Clarity about the nitric oxide (NO) concentrations existing physiologically is essential for developing a quantitative understanding of NO signalling, for performing experiments with NO that emulate reality, and for knowing whether or not NO concentrations become abnormal in disease states. A decade ago, a value of about 1 microM seemed ... More
Regulation of beta cell glucokinase by S-nitrosylation and association with nitric oxide synthase.
AuthorsRizzo MA, Piston DW
JournalJ Cell Biol
PubMed ID12707306
'Glucokinase (GK) activity plays a key role in glucose-stimulated insulin secretion from pancreatic beta cells. Insulin regulates GK activity by modulating its association with secretory granules, although little is known about the mechanisms involved in regulating this association. Using quantitative imaging of multicolor fluorescent proteins fused to GK, we found ... More
Constitutive and permissive roles of nitric oxide activity in embryonic ciliary cells.
AuthorsDoran SA, Tran CH, Eskicioglu C, Stachniak T, Ahn KC, Goldberg JI
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID12676758
'Embryos of Helisoma trivolvis exhibit cilia-driven rotation within the egg capsule during development. In this study we examined whether nitric oxide (NO) is a physiological regulator of ciliary beating in cultured ciliary cells. The NO donor S-nitroso-N-acetylpenicillamine (SNAP; 1-1,000 microM) produced a dose-dependent increase in ciliary beat frequency (CBF). In ... More
Direct evidence of nitric oxide presence within mitochondria.
'Nitric oxide (NO) has been implicated in the modulation of mitochondrial respiration, membrane potential, and subsequently in apoptosis. Although the presence of a mitochondrial NO synthase (mtNOS) has been described, there is no direct evidence in vivo of the presence of NO within mitochondria. It was the aim of this ... More
Neuronal nitric oxide synthase: its role and regulation in macula densa cells.
AuthorsKovács G, Komlósi P, Fuson A, Peti-Peterdi J, Rosivall L, Bell PD
JournalJ Am Soc Nephrol
PubMed ID14514725
'Macula densa (MD) cells detect changes in distal tubular sodium chloride concentration ([NaCl](L)), at least in part, through an apical Na:2Cl:K co-transporter. This co-transporter may be a site for regulation of tubuloglomerular feedback (TGF), and recently angiotensin II (Ang II) was shown to regulate the MD Na:2Cl:K co-transporter. In addition, ... More
Ceramide reduces endothelium-dependent vasodilation by increasing superoxide production in small bovine coronary arteries.
AuthorsZhang DX, Zou AP, Li PL
JournalCirc Res
PubMed ID11325875
'Ceramide serves as a second messenger in a variety of mammalian cells. Little is known regarding the role of ceramide in the regulation of vascular endothelial function. The present study was designed to determine whether ceramide affects endothelium-dependent vasodilation in coronary arteries and to explore the mechanism of action of ... More
Effect of hydrogen peroxide on guinea pig nasal mucosa vasculature.
AuthorsHirai T, Tsuru H, Tanimitsu N, Takumida M, Watanabe H, Yajin K, Sasa M
JournalJpn J Pharmacol
PubMed ID11202622
'The effect of hydrogen peroxide (H2O2) on guinea pig nasal mucosa vasculature was studied by in vitro assay. H2O2 elicited relaxation of guinea pig nasal mucosa strips precontracted with phenylephrine in a concentration-dependent manner. The relaxant response to H2O2 was abolished in the presence of catalase. Preincubation of the strips ... More
Formation of peroxynitrite in vascular endothelial cells exposed to cyclosporine A.
AuthorsNavarro-Antolin J, Lopez-Munoz MJ, Klatt P, Soria J, Michel T, Lamas S
JournalFASEB J
PubMed ID11282949
Methods for detection of reactive metabolites of oxygen and nitrogen: in vitro and in vivo considerations.
AuthorsTarpey MM, Wink DA, Grisham MB
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID14761864
Facile detection of reactive oxygen and nitrogen species in biologic systems is often problematic. This is a result of the numerous cellular mechanisms, both enzymatic and nonenzymatic involved in their catabolism/decomposition, the complex and overlapping nature of their reactivities, as well as the often limited intracellular access of detector systems. ... More
On the specificity of 4-amino-5-methylamino-2',7'-difluorofluorescein as a probe for nitric oxide.
AuthorsBalcerczyk A, Soszynski M, Bartosz G
JournalFree Radic Biol Med
PubMed ID15993331
The specificity of 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) for nitric oxide was evaluated in in vitro systems. The probe was found fairly specific for nitric oxide. Potential sources of artifacts include the autoxidation of DAF-FM, potentiated by light, and its oxidation by sources of superoxide and peroxyl radicals, leading to fluorescence spectra indistinguishable ... More
Nitric oxide inhibits adenosine 5'-triphosphate-induced Ca2+ response in inner hair cells of the guinea pig cochlea.
AuthorsShen J, Harada N, Yamashita T
JournalNeurosci Lett
PubMed ID12536042
To investigate the interaction between Ca(2+) and nitric oxide (NO) in inner hair cells of the guinea pig cochlea (IHCs), the extracellular adenosine 5'-triphosphate (ATP)-induced NO production and the effects of NO on ATP-induced increase of intracellular Ca(2+) concentrations ([Ca(2+)](i)) were investigated in IHCs using the NO-sensitive dye DAF-2 and ... More
Simultaneous measurement of intracellular Ca(2+) and nitric oxide: a new method.
AuthorsBerkels R, Dachs C, Roesen R, Klaus W
JournalCell Calcium
PubMed ID10859594
Different methods to measure the unstable radical nitric oxide (NO) have been established. We are going to present a new method to measure intracellular calcium and NO simultaneously in endothelial cells. A new fluorescent dye (DAF-2) has been developed recently which binds NO resulting in an enhanced fluorescence. We loaded ... More
Functional implications of Ca2+ mobilizing properties for nitric oxide production in aortic endothelium.
AuthorsKoyama T, Kimura C, Park SJ, Oike M, Ito Y
JournalLife Sci
PubMed ID12467891
We have investigated the relationship between Ca2+ mobilization and the cellular production of nitric oxide (NO) by using fura-2 and diaminofluorescein-2 (DAF-2), an NO-sensitive dye, in bovine aortic endothelial cells (BAEC). High concentrations of ATP (100 microM) or thapsigargin (1 micro M) depleted intracellular Ca2+ store sites with a single ... More
Ca2+ -linked upregulation and mitochondrial production of nitric oxide in the mouse preimplantation embryo.
AuthorsManser RC, Houghton FD
JournalJ Cell Sci
PubMed ID16638811
Previous studies have demonstrated a role for the signalling agent nitric oxide in regulating preimplantation embryo development. We have now investigated the biochemical mode of action of nitric oxide in mouse embryos in terms of mitochondrial function and Ca2+ signalling. DETA-NONOate, a nitric oxide donor, decreased day 4 blastocyst cell ... More
An iron-induced nitric oxide burst precedes ubiquitin-dependent protein degradation for Arabidopsis AtFer1 ferritin gene expression.
AuthorsArnaud N, Murgia I, Boucherez J, Briat JF, Cellier F, Gaymard F
JournalJ Biol Chem
PubMed ID16782706
Ferritins play an essential role in iron homeostasis by sequestering iron in a bioavailable and non-toxic form. In plants, ferritin mRNAs are highly and quickly accumulated in response to iron overload. Such accumulation leads to a subsequent ferritin protein synthesis and iron storage, thus avoiding oxidative stress to take place. ... More
Ca2+- and phosphatidylinositol 3-kinase-dependent nitric oxide generation in lung endothelial cells in situ with ischemia.
AuthorsAl-Mehdi AB, Song C, Tozawa K, Fisher AB
JournalJ Biol Chem
PubMed ID11042195
Endothelial cells generate nitric oxide (NO) in response to agonist stimulation or increased shear stress. In this study, we evaluated the effects of abrupt cessation of shear stress on pulmonary endothelial NO generation and its relationship to changes in intracellular Ca(2+). In situ endothelial generation of NO and changes in ... More
Neuroprotection of vestibular sensory cells from gentamicin ototoxicity obtained using nitric oxide synthase inhibitors, reactive oxygen species scavengers, brain-derived neurotrophic factors and calpain inhibitors.
AuthorsTakumida M, Anniko M, Shimizu A, Watanabe H
JournalActa Otolaryngol
PubMed ID12625566
OBJECTIVE: In order to devise a new treatment for inner ear disorders, the efficacy of a nitric oxide synthase inhibitor (L-N(G)-nitroarginine methylester [L-NAME]), a radical scavenger (D-methionine), a neurotrophin (brain-derived neurotrophic factor [BDNF]) and a calpain inhibitor (leupeptin) for protection from hair cell damage was investigated. MATERIAL AND METHODS: The ... More
Simultaneous in situ monitoring of intracellular Ca2+ and NO in endothelium of coronary arteries.
AuthorsYi FX, Zhang AY, Campbell WB, Zou AP, Van Breemen C, Li PL
JournalAm J Physiol Heart Circ Physiol
PubMed ID12388315
We developed an in situ assay system to simultaneously monitor intracellular Ca(2+) concentration ([Ca(2+)](i), fura 2 as indicator) and nitric oxide (NO) levels [4,5-diaminofluorescein as probe] in the intact endothelium of small bovine coronary arteries by using a fluorescent microscopic imaging technique with high-speed wavelength switching. Bradykinin (BK; 1 microM) ... More
Involvement of nitric oxide synthase in sucrose-enhanced hydrogen peroxide tolerance of Rhodococcus sp. strain APG1, a plant-colonizing bacterium.
AuthorsCohen MF, Yamasaki H
JournalNitric Oxide
PubMed ID14559426
Hydrogen peroxide (H2O2) tolerance of Rhodococcus sp. strain APG1, previously isolated from the aquatic fern Azolla pinnata, was examined in relation to nitric oxide (NO) production by cells cultured on a variety of C sources. Cells inoculated onto A. pinnata fronds established a surface-sterilant resistant density of 2-4x10(7) cells g(-1) ... More