Association Of Nitric Oxide Synthase Research Articles

Association of nitric oxide synthase 1 adaptor protein gene polymorphisms with schizophrenia in a Chinese Han population.

Introduction:Previous studies have analyzed the association between nitric oxide synthase 1 adaptor protein (NOS1AP) polymorphisms and schizophrenia; however, the results were inconsistent and there was a lack of evidence in a larger sample of Chinese Han population.Subjects and Methods:We decided to determine the association between four NOS1AP single-nucleotide polymorphisms (i.e., rs1858232A/G, rs4531275C/T, rs4657178C/T, and rs6704393C/T) and schizophrenia in northern Chinese Han population (350 patients and 522 controls) using restriction fragment length polymorphism.Results:Between schizophrenia group and healthy group, the genotype and allele frequencies for rs1858232A/G differed significantly (χ2 = 6.256, 4.145; P = 0.044, 0.045), but neither genotype nor allele frequencies of rs4531275C/T differed significantly. The genotype frequencies for rs4657178C/T and rs6704393C/T differed significantly (χ2 = 19.782, 12.683; P < 0.01, P = 0.002) between schizophrenia group and healthy group. In the gender-specific analysis, we found statistically significant difference in genotype frequencies between patients and controls in both subgroups for rs4657178C/T (χ2 = 9.356, 9.585; P = 0.009, 0.008). There was also a significant difference in the genotype frequency between patients and controls in male subgroup for rs6704393C/T (χ2 = 8.800, P = 0.012). In the haplotype analysis, only the TCT haplotype frequency of rs6704393C/T, rs4531275C/T, and rs4657178C/T differed significantly between patients and controls in total population (χ2 = 5.215, P = 0.022).In conclusion:Individuals with G allele of rs1858232A/G and C allele of rs4657178C/T which may be risk factors for schizophrenia should be given more attention, and also to individuals with the TCT haplotype, who are more likely to have schizophrenia. These results provide novel evidence for an association between NOS1AP polymorphisms and schizophrenia.

Disrupting the Interaction of nNOS with CAPON Prevents the Reinstatement of Morphine Conditioned Place Preference.

Drug abuse is a dramatic challenge for the whole society because of high relapse rate. Environmental cues are crucial for the preference memory of drug abuse. Extinction therapy has been developed to inhibit the motivational effect of drug cues to prevent the reinstatement of morphine abuse. However, extinction therapy alone only forms a new kind of unstable inhibitory memory. We found that morphine conditioned place preference (CPP) extinction training increased the association of nitric oxide synthase (nNOS) with its carboxy-terminal PDZ ligand (CAPON) in the dorsal hippocampus (dHPC) significantly and blocking the morphine-induced nNOS-CAPON association using Tat-CAPON-12C during and after extinction training reversed morphine-induced hippocampal neuroplasticity defect and prevented the reinstatement and spontaneous recovery of morphine CPP. Moreover, in the hippocampal selective ERK2 knock-out or nNOS knockout mice, the effect of Tat-CAPON-12C on the reinstatement of morphine CPP and hippocampal neuroplasticity disappeared, suggesting ERK2 is necessary for the effects of Tat-CAPON-12C. Together, our findings suggest that nNOS-CAPON interaction in the dHPC may affect the consolidation of morphine CPP extinction and dissociating nNOS-CAPON prevents the reinstatement and spontaneous recovery of morphine CPP, possibly through ERK2-mediated neuroplasticity and extinction memory consolidation, offering a new target to prevent the reinstatement of drug abuse.

Association between polymorphisms of NOS1, NOS2 and NOS3 genes and suicide behavior: a systematic review and meta-analysis.

The enzyme nitric oxide synthase has been associated with suicide behavior. NOS1, NOS2 and NOS3 genes are implicated in the production of nitric oxide. However, the association between NOS genes and suicide behavior has not yet been established. To assess the association of Nitric Oxide Synthase (NOS) genes and suicide behavior we performed a systematic review a meta-analysis. We searched articles published in three electronic databases, PubMed, Scopus and Web of Sciences, up to February 2019. We used keywords and combinations "NOS", "NOS1", "NOS2", "NOS3" and "suicide". Only articles that met the inclusion criteria were included. To assess the association between NOS genes and suicide behavior we used allelic, dominant and recessive models, as well as homozygous and heterozygous comparisons. The pooled results showed that rs2682826 of Nitric Oxide Synthase 1 gene (NOS1) increased the risk for suicide attempt in the allelic (OR: 1.34; 95 CI: 1.00-1.78), recessive (OR: 1.45; 95 CI:1.06-1.98) and heterozygous (OR: 1.41; 95 CI: 1.09-1.81) models. We found that the rs2682826 of NOS1 could increase the risk for suicide attempt. However, these results should only be taken as exploratory; more studies are necessary to determine the association between NOS genes and suicide behavior.

The effects of anti-hypertensive drugs and the mechanism of hypertension in vascular smooth muscle cell-specific ATP2B1 knockout mice

ATP2B1 is a gene associated with hypertension. We reported previously that mice lacking ATP2B1 in vascular smooth muscle cells (VSMC ATP2B1 KO mice) exhibited high blood pressure and increased intracellular calcium concentration. The present study was designed to investigate whether lack of the ATP2B1 gene causes a higher response to calcium channel blockers (CCBs) than to other types of anti-hypertensive drugs. Both VSMC ATP2B1 KO and control mice were administered anti-hypertensive drugs while monitoring blood pressure shifts. We also examined the association of nitric oxide synthase (NOS) activity in those mice to investigate whether another mechanism of hypertension existed. VSMC ATP2B1 KO mice exhibited significantly greater anti-hypertensive effects with a single injection of nicardipine, but the effects of an angiotensin II receptor blocker (ARB), an α-blocker and amlodipine on blood pressure were all similar to control mice. However, long-term treatment with amlodipine, but not an ARB, significantly decreased the blood pressure of KO mice compared with control mice. Both mRNA and protein expression levels of the L-type calcium channel were significantly upregulated in KO VSMCs. There were no alterations in neural NOS protein expression of VSMCs or in urinary NO production between the two groups. VSMC ATP2B1 KO mice had a higher response to CCBs for blood pressure-lowering effects than other anti-hypertensive drugs. These results mean that increased intracellular calcium concentration in VSMCs due to lack of ATP2B1 and subsequent activation of L-type calcium channels mainly affects blood pressure and suggests increased susceptibility to CCBs in this type of hypertension.

Association of nitric oxide synthase 3 gene polymorphism with the risk of type 2 diabetes.

Type-2 diabetes (T2D) is a multifactorial (environmental and genetic factors) and global epidemic disease with an estimated high prevalence worldwide. Studies have indicated that nitric oxide synthase 3 (NOS3) has several important roles in the pathogenesis of T2D. The present study aims to investigate the association between NOS3 rs1800779(A/G) and T2D in an Iranian sample population. A case-control study was conducted on 250 T2D patients and 250 healthy control subjects (HCs). Genotyping of the rs1800779(A/G) variant was conducted using a Tetra-Amplification Refractory Mutation System polymerase chain reaction. The frequencies of genotypes AA, AG and GG polymorphisms were 56.8, 39.2 and 4% in the T2D group, and 42.8, 56 and 1.2% in the HCs group, respectively. The frequency of the minor (G) allele was 23.6% in the T2D group and 29.2% in the HCs group. The genotype frequencies of the rs1800779(A/G) variant demonstrated statistically significant differences between T2D and controls in a codominant model (AG vs. AA, OR=0.527, 95% CI=0.368-0.756, P<0.001) and dominant model (AG+GG vs. AA, OR=0.569, 95% CI=0.399-0.811, P=0.002). There was no significant association between clinical and demographic characteristics and the NOS3 rs1800779(A/G) polymorphism in dominant status (P>0.05). The dominant model and AG genotype of NOS3 rs1800779(A/G) polymorphism may had a protective effect on T2D of Iranian population.

Role of polymorphisms of inducible nitric oxide synthase and endothelial nitric oxide synthase in idiopathic environmental intolerances.

Oxidative stress and inflammation play a pathogenetic role in idiopathic environmental intolerances (IEI), namely, multiple chemical sensitivity (MCS), fibromyalgia (FM), and chronic fatigue syndrome (CFS). Given the reported association of nitric oxide synthase (NOS) gene polymorphisms with inflammatory disorders, we aimed to investigate the distribution of NOS2A −2.5 kb (CCTTT)n as well as Ser608Leu and NOS3 −786T>C variants and their correlation with nitrite/nitrate levels, in a study cohort including 170 MCS, 108 suspected MCS (SMCS), 89 FM/CFS, and 196 healthy subjects. Patients and controls had similar distributions of NOS2A Ser608Leu and NOS3 −786T>C polymorphisms. Interestingly, the NOS3 −786TT genotype was associated with increased nitrite/nitrate levels only in IEI patients. We also found that the NOS2A −2.5 kb (CCTTT)11 allele represents a genetic determinant for FM/CFS, and the (CCTTT)16 allele discriminates MCS from SMCS patients. Instead, the (CCTTT)8 allele reduces by three-, six-, and tenfold, respectively, the risk for MCS, SMCS, and FM/CFS. Moreover, a short number of (CCTTT) repeats is associated with higher concentrations of nitrites/nitrates. Here, we first demonstrate that NOS3 −786T>C variant affects nitrite/nitrate levels in IEI patients and that screening for NOS2A −2.5 kb (CCTTT)n polymorphism may be useful for differential diagnosis of various IEI.

Association of nitric oxide synthase 3 (NOS3) 894 G&gt;T polymorphism with prognostic outcomes of anthracycline in Chinese patients with acute myeloid leukaemia

The aim of the present study was to investigate the influence of the nitric oxide synthase 3 (NOS3) 894G>T polymorphism on prognostic outcomes of anthracycline in Chinese patients with de novo intermediate-risk acute myeloid leukaemia (AML) and to examine the gene expression level in relation to genetic variation. In all, 225 Chinese patients with intermediate-risk AML (at the complete remission stage) treated with anthracycline were enrolled in the study. The 894G>T polymorphism of the NOS3 gene was analysed by allele-specific matrix-assisted laser desorption ionization time-of-flight. Expression of NOS3 mRNA was tested in 72 patients of known genotype for NOS3 894G>T. The clinical characteristics of these patients were obtained from medical records. Survival analysis showed that patients with AML (GG genotype) had a longer overall survival (OS; P=0.006). After adjusting for age, gender, leucocyte count, haemoglobin level, platelet level, French, American and Britain (FAB) classification, lactate dehydrogenase levels, Eastern Cooperative Oncology Group Performance Status, nucleophosmin gene and fms-related tyrosine kinase 3 gene, multivariate survival analysis showed that the NOS3 894G>T polymorphism appeared to be a predicting factor for OS (P=0.014; hazard ratio=1.856). However, no significant associations between the NOS3 894G>T polymorphism and relapse-free survival and relapse in patients with AML were observed. Gene expression levels were significantly higher in patients with the GG genotype than in patients with the GT and TT genotypes (P=0.033). The findings suggest that the NOS3 894G>T variant may be a biomarker for the prediction of OS in Chinese patients with AML.

Rebuttal: The independent association of nitric oxide synthase 3 polymorphisms and their combination with inflammatory markers

Rebuttal: The independent association of nitric oxide synthase 3 polymorphisms and their combination with inflammatory markers -

Enhancement of nitric oxide production by association of nitric oxide synthase with N‐methyl‐d‐aspartate receptors via postsynaptic density 95 in genetically engineered Chinese hamster ovary cells: real‐time fluorescence imaging using nitric oxide sensitive dye

The current quantitative study demonstrates that the recruitment of neuronal nitric oxide synthase (nNOS) beneath N-methyl-D-aspartate (NMDA) receptors, via postsynaptic density 95 (PSD-95) proteins significantly enhances nitric oxide (NO) production. Real-time single-cell fluorescence imaging was applied to measure both NO production and Ca(2+) influx in Chinese hamster ovary (CHO) cells expressing recombinant NMDA receptors (NMDA-R), nNOS, and PSD-95. We examined the relationship between the rate of NO production and Ca(2+) influx via NMDA receptors using the NO-reactive fluorescent dye, diaminofluorescein-FM (DAF-FM) and the Ca(2+)-sensitive yellow cameleon 3.1 (YC3.1), conjugated with PSD-95 (PSD-95-YC3.1). The presence of PSD-95 enhanced the rate of NO production by 2.3-fold upon stimulation with 100 microm NMDA in CHO1(+) cells (expressing NMDA-R, nNOS and PSD-95) when compared with CHO1(-) cells (expressing NMDA-R and nNOS lacking PSD-95). The presence of nNOS inhibitor or NMDA-R blocker almost completely suppressed this NMDA-stimulated NO production. The Ca(2+) concentration beneath the NMDA-R, [Ca(2+)](NR), was determined to be 5.4 microm by stimulating CHO2 cells (expressing NMDA-R and PSD-95-YC3.1) with 100 microm NMDA. By completely permealizing CHO1 cells with ionomycin, a general relationship curve of the rate of NO production versus the Ca(2+) concentration around nNOS, [Ca(2+)](NOS), was obtained over the wide range of [Ca(2+)](NOS). This sigmoidal curve had an EC(50) of approximately 1.2 microm of [Ca(2+)](NOS), implying that [Ca(2+)](NR) = 5.4 microm can activate nNOS effectively.

Association of nitric oxide synthase 2(NOS2G-954C) polymorphism with cardiomyopathy in Trypanosoma cruzi infection

Association of nitric oxide synthase 2(NOS2G-954C) polymorphism with cardiomyopathy in Trypanosoma cruzi infection

Migraine association and linkage studies of an endothelial nitric oxide synthase (NOS3) gene polymorphism.

Migraine shows strong familial aggregation. However, the number of genes involved in the disorder is unknown and not identified. Nitric oxide is involved in the central processing of pain stimuli and plays an important role in the regulation of basal or stimulated vasodilation. Nitric oxide synthase, which controls the synthesis of nitric oxide, could possibly be a cause, or candidate gene, in migraine etiology. In this study, we detected a polymorphism for endothelial nitric oxide synthase by polymerase chain reaction and tested this for association and linkage to migraine. Results from the study did not show an association of the nitric oxide synthase microsatellite when tested in 91 affected and 85 unaffected individuals. Using the FASTLINK program for parametric linkage analysis, the polymorphism did not show significant linkage to migraine when tested in four migraine pedigrees composed of 116 individuals, 52 affected. Total LOD scores excluded linkage up to 8.5 cM between the nitric oxide synthase polymorphism and migraine. Results using the nonparametric affected pedigree member form of analysis also did not support a role for this gene in migraine etiology.

Nitric oxide synthase (NOS) I during postnatal development in rat and mouse skeletal muscle

Previous studies on adult rat and mouse skeletal muscles have shown the spatial association of nitric oxide synthase (NOS) I to the dystrophin complex (DC) in the sarcolemma of type II fibers and, in combination with the NMDA receptor-1 (NMDAR-1), an accumulation of the enzyme at the neuromuscular junctions (NMJ) of this fiber type. Using immunohistochemistry, enzyme histochemistry and alpha-bungarotoxin labeling we report here temporal relationships of NOS I, members of the DC, other components of the cortical cytoskeleton in the junctional and non-junctional sarcolemma as well as of molecules involved in NMJ transmission of either type I or II myofibers especially in head and neck muscles during postnatal rat and mouse development. Fiber typing was performed by specific anti-myosin antibodies. Beginning with postnatal day (PD) 1 in both fiber types dystrophin, dystrophin-associated glycoproteins (DAG), beta-dystroglycan, alpha-sarcoglycan (adhalin) and spectrin were present in the junctional and extrajunctional sarcolemma, while utrophin, acetylcholinesterase, alpha-bungarotoxin labeled acetylcholine receptors were concentrated in the NMJ of both fiber types. NOS I activity and immunoreactivity were only found in the NMJ region of type II fibers, where NMDAR-1 appeared around PD 15. Primarily in the tongue there was no strict correlation between muscle fiber type and NOS I behaviour during early postnatal development, and muscle fibers not reactive for myosin antibodies against both fiber types were negative or positive for NOS I but always positive for the other molecules either in both the junctional and extrajunctional sarcolemma or in the NMJ only; later all muscle fibers of the tongue were of type II and NOS I-positive. Maturation of enzyme activities, immunoreactivities and AChR intensity depended on the respective muscle and can last until PD 50; in the tongue and neck muscles they appeared to increase approximately until PD 20 or 25. In conclusion, in type II fibers of rat and mouse skeletal muscle all molecules with the exception of NMDAR-1 and relevant for NOS I targeting and positioning as well as function inside and outside the NMJ are already present at birth, but their concentrations and/or activities increase postnatally, and the adult situation appears to be reached between the third and seventh week of postnatal life. Therefore, initial interactions between NOS I and the other molecules necessary for the formation of the NOS I-DC in and on the way to the sarcolemma presumably take place before birth.

Vesicle membrane association of nitric oxide synthase in primary mouse macrophages.

The isoform of nitric oxide synthase (NOS) whose activity is independent of elevated Ca2+ and exogenous calmodulin (iNOS; NOS type II) is inducible in a wide variety of cells and plays a major role in pathophysiology. The notion that iNOS is predominantly cytosolic is based on studies of a transformed cell line; almost nothing is known about the subcellular localization of iNOS in primary cells. Accordingly, we undertook a combined immunoelectron microscopic and biochemical analysis of iNOS in primary mouse macrophages. Approximately one-half of their iNOS activity and protein could be sedimented from 1 M KCl at 100,000 x g. The morphologic counterpart of particulate iNOS was a population of 50 to 80 nm vesicles that did not correspond to lysosomes nor peroxisomes. Vesicular iNOS arose from cytosolic iNOS by undergoing a post-translational modification that increased its apparent molecular mass by 4.5 kDa and promoted its salt-, detergent-, acid- and urea-resistant association with membranes, in the absence of detectable alternative splicing, myristoylation, palmitoylation, acetylation, glycosylation, or COOH-terminal truncation. Although primary macrophage iNOS underwent phosphorylation, ubiquitinylation, and binding of calmodulin tightly enough to resist boiling in SDS, these modifications did not allow us to distinguish between the cytosolic and particulate variants. The apparently novel iNOS-positive vesicles may translocate to phagosomes containing appropriately opsonized particles.