Melatonin-Nitric Oxide Crosstalk in Plants and the Prospects of NOMela as a Nitric Oxide Donor.

Melatonin regulates vital physiological processes in animals, such as the circadian cycle, sleep, locomotion, body temperature, food intake, and sexual and immune responses. In plants, melatonin modulates seed germination, longevity, circadian cycle, photoperiodicity, flowering, leaf senescence, postharvest fruit storage, and resistance against biotic and abiotic stresses. In plants, the effect of melatonin is mediated by various regulatory elements of the redox network, including RNS and ROS. Similarly, the radical gas NO mediates various physiological processes, like seed germination, flowering, leaf senescence, and stress responses. The biosynthesis of both melatonin and NO takes place in mitochondria and chloroplasts. Hence, both melatonin and nitric oxide are key signaling molecules governing their biological pathways independently. However, there are instances when these pathways cross each other and the two molecules interact with each other, resulting in the formation of N-nitrosomelatonin or NOMela, which is a nitrosated form of melatonin, discovered recently and with promising roles in plant development. The interaction between NO and melatonin is highly complex, and, although a handful of studies reporting these interactions have been published, the exact molecular mechanisms governing them and the prospects of NOMela as a NO donor have just started to be unraveled. Here, we review NO and melatonin production as well as RNS-melatonin interaction under normal and stressful conditions. Furthermore, for the first time, we provide highly sensitive, ozone-chemiluminescence-based comparative measurements of the nitric oxide content, as well as NO-release kinetics between NOMela and the commonly used NO donors CySNO and GSNO.

Products of oxidative and non-oxidative metabolism of L-arginine as potential regulators of Ca2+ transport in mitochondria of uterine smooth muscle.

Mitochondria play a crucial role in maintaining Ca2+ homeostasis in cells. Due to the critical regulatory role of the products of oxidative and non-oxidative metabolism of L-arginine, it is essential to clarify their effect on Ca2+ transport in smooth muscle mitochondria. Experiments were performed on the uterine myocytes of rats and isolated mitochondria. The possibility of NO synthesis by mitochondria was demonstrated by confocal microscopy and spectrofluorimetry methods using the NO-sensitive fluorescent probe DAF-FM and Mitotracker Orange CM-H2TMRos. It was shown that 50 µM L-arginine stimulates the energy-dependent accumulation of Ca2+ in mitochondria using the fluorescent probe Fluo-4 AM. A similar effect occurred when using nitric oxide donors 100 µM SNP, SNAP, and sodium nitrite (SN) directly. The stimulating effect was eliminated in the presence of the NO scavenger C-PTIO. Nitric oxide reduces the electrical potential in mitochondria without causing them to swell. The stimulatory effect of spermine on the accumulation of Ca2+ by mitochondria is attributed to the enhancement of NO synthesis, which was demonstrated with the use of C-PTIO, NO-synthase inhibitors (100 µM NA and L-NAME), as well as by direct monitoring of NO synthesis fluorescent probe DAF-FM. A conclusion was drawn about the potential regulatory effect of the product of the oxidative metabolism of L-arginine - NO on the transport of Ca2+ in the mitochondria of the myometrium, as well as the corresponding effect of the product of non-oxidative metabolism -spermine by increasing the synthesis of NO in these subcellular structures.

DNIC is a Structure Providing Specificity of Physiological Effects of NO.

Metabolism of nitric oxide (NO) donors: dinitrosyl iron complexes (DNIC), nitrosothiols (RSNO), and nitroprusside was studied on a chick embryo model. The obtained results give reason to assume that DNIC constituting the main pool of nitroso compounds in the vast majority of tissues are NO donors immediately interacting with the physiological target of NO, and other NO donors can perform this function after their transformation into DNIC. NO is released from DNIC not spontaneously, but under a joint influence of a factor destroying the complex and a target having chemical affinity for NO. A similar mechanism is apparently implicated in NO passage through the cell membrane.

Role of NO in arterial vascular function of intertidal fish (Girella laevifrons) and marine fish (Isacia conceptionis) / Papel do NO na função vascular arterial de peixes entremarés (Girella laevifrons) e peixes marinhos (Isacia conceptionis)

Abstract Previous studies performed in intertidal fish (Girella laevifrons),as well as marine fish (Isacia conceptionis), showed that acetylcholine (ACh) produced contractions mediated by cyclooxygenases that were dependent on the area and potency of contraction in several arterial vessels. Given that the role of nitric oxide is poorly understood in fish, the objective of our study was to evaluate the role of nitric oxide in branchial afferent (ABA), branchial efferent (ABE), dorsal (DA) and mesenteric (MA) arterial vessels from both Girella laevifrons and Isacia conceptionis. We studied afferent and efferent branchial, dorsal and mesenteric arteries that were dissected from 6 juvenile specimens. Isometric tension studies were done using dose response curves (DRC) for Ach (10–13 to 10–3 M) and blockade with L-NAME (10–5 M), and DRC for sodium nitroprusside (SNP, a donor of NO). L-NAME produced an attenuation of the contractile response in the dorsal, afferent and efferent branchial arteries and a potentiation of the contraction in the MA. SNP caused 70% dilation in the mesenteric artery and 40% in the dorsal artery. Our results suggest that Ach promotes precarious dilatation in MA mediated by NO; data that is supported by the use of sodium nitroprusside. In contrast, in the vessels DA, ABA and EBA our results support that the pathway Ach-NO-relaxation is absent in both species.

Resumo Estudos anteriores, realizados no peixe intertidal (Girellalaevifrons) no peixe marinho (Isacia conceptionis), mostram que a acetilcolina (Ach) provoca contrações mediadas por ciclooxigenases que eram dependentes da área e potencia da contração em vários vasos arteriais. Tendo em conta que o papel do óxido nítrico é mal compreendido em peixes, o objetivo do nosso estudo foi avaliar o papel do óxido nítrico em vasos arteriais de ambos os peixes Girella laevifrons e Isacia conceptionis. Nós estudamos os vasos aferente, branquial (ABA), eferente branquial (ABE), dorsal (DA) e mesentérica (MA), que foram dissecadas de seis espécimes juvenis. Estudos de tensão isométrica foram realizados utilizando as curvas de dose-resposta (DRC) para Ach (10–13 a 10–3M) e bloqueio com L-NAME (10–5 M), e na DRC para o nitroprussiato de sódio (SNP, doador do NO). L- NAME produziu uma atenuação da resposta contrátil nas artérias dorsais, aferentes e eferentes branquial e uma potenciação da contração no MA. SNP causaram 70% da dilatação da artéria mesentérica e 40% na artéria dorsal. Nossos resultados sugerem que Ach promove dilatação precária em MA mediada por NO; dados que é suportada pela utlilização de nitroprussiato de sódio. Em contraste, nos vasos de DA, ABA e EBA nossos resultados suportam que a via de Ach-NO-relaxamento está ausente em ambas as espécies.

Nitric oxide synthesis and biological functions of nitric oxide released from ruthenium compounds

During three decades, an enormous number of studies have demonstrated the critical role of nitric oxide (NO) as a second messenger engaged in the activation of many systems including vascular smooth muscle relaxation. The underlying cellular mechanisms involved in vasodilatation are essentially due to soluble guanylyl-cyclase (sGC) modulation in the cytoplasm of vascular smooth cells. sGC activation culminates in cyclic GMP (cGMP) production, which in turn leads to protein kinase G (PKG) activation. NO binds to the sGC heme moiety, thereby activating this enzyme. Activation of the NO-sGC-cGMP-PKG pathway entails Ca2+ signaling reduction and vasodilatation. Endothelium dysfunction leads to decreased production or bioavailability of endogenous NO that could contribute to vascular diseases. Nitrosyl ruthenium complexes have been studied as a new class of NO donors with potential therapeutic use in order to supply the NO deficiency. In this context, this article shall provide a brief review of the effects exerted by the NO that is enzymatically produced via endothelial NO-synthase (eNOS) activation and by the NO released from NO donor compounds in the vascular smooth muscle cells on both conduit and resistance arteries, as well as veins. In addition, the involvement of the nitrite molecule as an endogenous NO reservoir engaged in vasodilatation will be described.

Respuesta de las arterias pulmonares de lechón al óxido nítrico y a dadores de óxido nítrico: Evolución con la edad postnatal y modulación por el estrés oxidativo / Nitric oxide- and nitric oxide donor-induced responses in piglet pulmonary arteries: Postnatal maturation and modulation by oxidative stress

En el presente estudio hemos analizado la influencia de la edad postnatal y los cambios en el nivel de estrés oxidativo sobre la vasodilatación pulmonar in vitro inducida por el NO endógeno, el NO exógeno y donadores de NO. Se han utilizado las arterias pulmonares procedentes de lechones de 1 día y de 2 semanas de edad para el registro de la fuerza contráctil. En las arterias pulmonares de lechones de 1 y 15 días de edad, el estrés oxidativo basal modula la acción vasodilatadora del NO, siendo la NAD(P)H oxidasa de la adventicia la principal fuente endógena del anión superóxido. La vasodilatación inducida por el NO de origen endotelial y por el NO exógeno aumenta con la edad, posiblemente por un incremento en la actividad de la ciclooxigenasa-1 en los primeros momentos de vida extrauterina que modularía el efecto vasodilatador del NO. Finalmente, encontramos que el NO y los donadores de NO, SNAP y nitroprusiato (SNP), difieren en la cinética y distribución regional de la liberación del NO, lo que influye en la susceptibilidad para la inactivación por el anión superóxido y por la oxihemoglobina (AU)