Nitric oxide: a synchronizing chemical messenger

Nitric oxide (NO) has been recognized as a ubiquitous chemical messenger in a large number of different biological systems. Its chemical properties make it less specific and less controllable than practically any other neurotransmitter or hormone. In view of this, its extensive biological role as a chemical messenger seems surprising. It is suggested that the biological function of NO evolved early in the anaerobic stage of evolution. In view of its low molecular weight, limited interaction with water, and its electrical neutrality, which allow it to diffuse rapidly through the cytoplasm and biomembranes, it is suggested that the need for NO has been retained by and maintained in eukaryote cells because of its ability to affect many biochemical functions simultaneously, acting primarily as an intracellular synchronizing chemical messenger.     

Nitric oxide biosynthesis, nitric oxide synthase inhibitors and arginase competition for L-arginine utilization

Nitric oxide (NO) is a recently discovered mediator produced by mammalian cells. It plays a key role in neurotransmission, control of blood pressure, and cellular defense mechanisms. Nitric oxide synthases (NOSs) catalyze the oxidation of L-arginine to NO and L-citrulline. NOSs are unique enzymes in that they possess on the same polypeptidic chain a reductase domain and an oxygenase domain closely related to cytochrome P450s. NO and superoxide formation as well as NOS stability are finely regulated by Ca²⁺/calmodulin interactions, by the cofactor tetrahydrobiopterin, and by substrate availability. Strong interactions between the L-arginine-metabolizing enzymes are clearly demonstrated by competition between NOSs and arginases for L-arginine utilization, and by potent inhibition of arginase activity by N^ω-hydroxy-L-arginine, an intermediate in the L-arginine to NO pathway.     

Nitric oxide synthase reduces nitrite to NO under anoxia

Cultured bEND.3 endothelial cells show a marked increase in NO production when subjected to anoxia, even though the normal arginine pathway of NO formation is blocked due to absence of oxygen. The rate of anoxic NO production exceeds basal unstimulated NO synthesis in normoxic cells. The anoxic release of NO is mediated by endothelial nitric oxide synthase (eNOS), can be abolished by inhibitors of NOS and is accompanied by consumption of intracellular nitrite. The anoxic NO release is unaffected by the xanthine oxidase inhibitor oxypurinol. The phenomenon is attributed to anoxic reduction of intracellular nitrite by eNOS, and its magnitude and duration suggests that the nitrite reductase activity of eNOS is relevant for fast NO delivery in hypoxic vascular tissues. Received 20 August 2006; received after revision 21 September 2006; accepted 8 November 2006     

Nitric oxide inhibition intensifies cold-restraint induced gastric ulcers in rats

Treatment 20 min beforehand with an inhibitor of nitric oxide (NO) synthesis, N^W-nitro-l-arginine methyl ester (L-NAME) (12.5, 25, 50 or 100 mg/kg, s.c.), dose-dependently intensified gastric glandular mucosal ulceration produced by cold-restraint stress. Hexamethonium (20 mg/kg) or atropine (1 mg/kg) pretreatment s.c. 20 min before stress strongly antagonised stress-evoked ulceration, as well as the ulcer-potentiating effects of L-NAME when either cholinoceptor antagonist was given concurrently with the NO inhibitor. Stress-induced mast cell degranulation was not worsened by L-NAME pretreatment. The findings suggest that NO could confer partial protection against stress-induced gastric ulcer formation; its activity is triggered off by the ulcerogenic mechanism of stress.     

Nitric oxide can inhibit apoptosis or switch it into necrosis

Nitric oxide (NO) and its related molecules are important messengers that play central roles in pathophysiology. Redox modulation of thiol groups on protein cysteine residues by S-nitrosylation can modulate protein function. NO has emerged as a potent regulator of apoptosis in many cell types, either preventing cell death or driving an apoptotic response into a necrotic one. NO protects neuroblastoma cells from retinoid- and cisplatin-induced apoptosis, without significantly increasing necrotic cell damage. Nitrosylation of thiol groups of several critical factors may be important for cell survival. Indeed, S-nitrosylation of the active-site cysteine residue of apoptotic molecules, such as caspases and tissue transglutaminase, results in the inhibition of their catalytic activities and has important implications for the regulation of apoptosis by NO. On the other hand, NO is able to shift the anti-CD95- and ceramide-triggered apoptotic response of Jurkat T cells into necrotic cell death. In these apoptotic models, NO is therefore unable to solely inhibit cell death, indicating that it may act below the point of no return elicited by CD95-ligation and ceramide stimulation.     

Nitric oxide synthases: targets for therapeutic strategies in neurological diseases

Glutamate excitotoxicity, oxidative stress, and mitochondrial dysfunctions are common features leading to neuronal death in cerebral ischemia, traumatic brain injury, Parkinson's disease, Huntington's disease, Alzheimer's disease and amyotrophic lateral sclerosis. Nitric oxide (NO) alone or in cooperation with superoxide anion and peroxynitrite is emerging as a predominant effector of neurodegeneration The use of NO synthase (NOS) inhibitors and mutant mice lacking each NOS isoform have provided evidence for the injurious effects of NO derived from neuronal or inducible isoforms. New neuroprotective strategies have been proposed with selective NOS inhibitors for the neuronal (ARL17477) or the inducible (1400 W) isoforms or with compounds combining in one molecule selective nNOS inhibition and antioxidant properties (BN 80933), in experimental ischemia-induced acute neuronal damage. The efficacy of these new strategies is well established in acute neuronal injury but remains to be determined in more chronic neurological diseases.     

Nitric oxide stimulates human neural progenitor cell migration via cGMP-mediated signal transduction

Neuronal migration is one of the most critical processes during early brain development. The gaseous messenger nitric oxide (NO) has been shown to modulate neuronal and glial migration in various experimental models. Here, we analyze a potential role for NO signaling in the migration of fetal human neural progenitor cells. Cells migrate out of cultured neurospheres and differentiate into both neuronal and glial cells. The neurosphere cultures express neuronal nitric oxide synthase and soluble guanylyl cyclase that produces cGMP upon activation with NO. By employing small bioactive enzyme activators and inhibitors in both gain and loss of function experiments, we show NO/cGMP signaling as a positive regulator of migration in neurosphere cultures of early developing human brain cells. Since NO signaling regulates cell movements from developing insects to mammalian nervous systems, this transduction pathway may have evolutionary conserved functions.     

Nitric oxide: a radical molecule in quest of free radicals in proteins

A number of enzymes use an amino acid free radical cofactor. Tyrosyl and tryptophanyl radicals react with nitric oxide (NO) with an almost diffusion-limited rate. The catalytically competent tyrosyl radical in ribonucleotide reductase (RR) and prostaglandin H synthase (PGHS) recombines with NO in a radical-radical reaction. The unstable adduct formed can dissociate to regenerate the tyrosyl radical. However, upon prolonged incubation with NO, the diiron center of mouse RR leaks out, while the adduct is sucessively oxidized into an iminoxyl radical and a nitrotyrosine in PGHS. These data provide a plausible mechanism for the physiological inactivation of RR observed in various models, and may help in understanding the inhibition of PGHS reported in some cases. Reversible combination with NO is an intrinsic property of tyrosyl radicals, which also occurs with Y_D ^• and Y_Z ^• in photosystem II, where NO has been useful in the analysis of the oxygen-evolving complex.     

Control of respiration by nitric oxide in Keilin-Hartree particles,mitochondria and SH-SY5Y neuroblastoma cells

The pattern of cytochrome c oxidase inhibition by nitric oxide (NO) was investigated polarographically using Keilin-Hartree particles, mitochondria and human neuroblastoma cells. NO reacts with purified cytochrome c oxidase forming either a nitrosyl- or a nitrite-inhibited derivative, displaying distinct kinetics and light sensitivity of respiration recovery in the absence of free NO. Keilin-Hartree particles or cells, respiring either on endogenous substrates alone or in the presence of ascorbate, as well as state 3and state 4mitochondria respiring on glutamate and malate, displayed the rapid recovery characteristic of the nitrite derivative. All systems, when respiring in the presence of tetramethyl-p-phenylenediamine, were characterised by the slower, light-sensitive recovery typical of the nitrosyl derivative. Together the results suggest that the reaction of NO with cytochrome c oxidase in situ follows two alternative inhibition pathways, depending on the electron flux through the respiratory chain.Received 1 April 2003; received after revision 22 May 2003; accepted 3 June 2003     

Aerial and aquatic respiration of certain trochids

Résumé La consommation d'oxygène de 4 espèces de Troques dans l'air et dans l'eau est en relation avec leur distribution dans la zone intercotidale.

---

---

I thank ProfessorG. E. Newell for his advice.     

Ultradian and other rhythms in human respiration rate

Summary Twelve male subjects had their respiration rate (RR) measured at 3 min intervals for 6 h. Although there were substantial individual differences, most subjects demonstrated a 90±15 min rhythm of RR, with several subjects also showing 60±10 min and 30±3 min rhythms.     

Inhibition of nuclear and mitochondrial respiration by arsenite

Résumé La respiration des noyaux isolés du thymus de rat est inhibée par l'arsénite à une concentration 10 fois supérieure à celle qui inhibe la respiration mitochondriale. Ce fait suggère, une difficulté de pénétration à travers la membrane nucleaire et ne rend pas compte de la différence entre le méchanisme de la phosphorylation oxydative des noyaux et celle des mitochondria.     

Stimulation by D-glucose of mitochondrial respiration

Summary D-glucose increases O₂ uptake by cerebellum mitochondria. This effect is abolished by D-glucose-6-phosphate and D-mannoheptulose. It is proposed that the phosphorylation of D-glucose as catalyzed by bound hexokinase directly affects mitochondrial respiration.     

A comparative approach to fish respiration

Zusammenfassung Es wird ein Überblick über neuere Forschungen gegeben, die das weite Arbeitsgebiet des Mechanismus der Fischatmung betreffen. Nachdruck wird auf den vergleichenden Standpunkt einer solchen Forschungsarbeit gelegt, wobei gezeigt wird, dass die Wahl besonderer Spezies ein förderndes Licht auf allgemein wichtige Probleme werfen kann, die bei Kiemenuntersuchungen sowohl auf der Makrostrukturstufe wie auf der Ultrastrukturstufe auftreten. Vom physiologischen Standpunkt aus gesehen, erweisen sich einige Arten auf Grund ihrer Lebensweise für experimentelle Untersuchungen ihres Sauerstoffverbrauchs besonders geeignet. Die strukturelle Anordnung des Pumpenmechanismus anderer Arten erleichert wiederum äusserst günstig die Direktmessung des Ventilationsvolumens und anderer mit dem Gasaustausch zusammenhängender Parameter.

---

---

Based on lectures given in Spring 1969 at Duke University (N. Carolina, USA), Universities of Goteborg, Umca and Uppsala (Sweden).     

Stimulation by D-glucose of mitochondrial respiration

D-glucose increases O2 uptake by cerebellum mitochondria. This effect is abolished by D-glucose-6-phosphate and D-mannoheptulose. It is proposed that the phosphorylation of D-glucose as catalyzed by bound hexokinase directly affects mitochondrial respiration.     

Oscillatory respiration in balanus amphitrite darwin

Résumé ChezBalanus amphitrite, espèce des eaux chaudes, l'utilisation de l'oxygène par des individus isolés (extraits de leur coquille), soumis à une température peu élevée est de forme oscillatoire, contrairement à ce que l'on observe chez des espèces des eaux froides. Il se peut que cette différence caractérise de manière générale deux types d'espèces.     

Nitric oxide mediates histamine induced down-regulation of H<Subscript>2</Subscript> receptor mRNA and internalization of the receptor protein (R1)

During agonist-dependent long-term stimulation of cells, histamine receptor subtypes are frequently down-regulated. However, the mechanisms underlying the modulation of receptor expression during long-term histamine stimulation have yet to be resolved. Based on our recently reported results showing an H₁-mediated down-regulation of histamine H₂ receptor mRNA in endothelial cells, our aim was to characterize the mechanism controlling rapid and long-term histamine-mediated modulation of H₂ receptor expression in more detail. We were able to show that the histamine-induced down-regulation of H₂ receptor mRNA and cell surface expression lasting for 24 h was accompanied by augmentation of the receptor protein level in the cytoplasmatic fraction of endothelial cells for this time period. Furthermore, changes in receptor protein levels in whole-cell lysate were negligible, indicating that the rapid and prolonged modulation of cell surface H₂ receptor levels by histamine was regulated solely via internalization. The role of nitric oxide (NO) as a key mediator in histamine-stimulated cell responses was underlined by subsequent studies showing the attenuation of histamine-induced H₂ receptor mRNA down-regulation and protein trafficking following NO synthase isozyme inhibition.Received 11 March 2003; received after revision 11 June 2003; accepted 17 June 2003     

Temperature-controlled alternative respiration and outgrowth rate from conidia ofNeurospora crassa

Summary Incubation of conidia of wild typeNeurospora crassa at temperatures ranging from 25 to 46°C modulates their respiratory type. Between 37 and 41°C, the transient activity of the cyanide-insensitive respiratory pathway parallels, with a maximal extrusion of protons into the medium, the optimal rate of germ tube outgrowth.We are grateful to Mrs Arlette Cattaneo for her competent technical assistance.