Ceramide in apoptosis signaling: relationship with oxidative stress

Ceramide is one of the major sphingosine-based lipid second messengers that is generated in response to various extracellular agents. However, while widespread attention has focused on ceramide as a second messenger involved in the induction of apoptosis, important issues with regard to the mechanisms of ceramide formation and mode of action remain to be addressed. Several lines of evidence suggest that ceramide and oxidative stress are intimately related in cell death induction. This review focuses on the putative relationships between oxidative stress and sphingolipid metabolism in the apoptotic process and discusses the potential mechanisms that connect and regulate the two phenomena.     

Neuregulins: functions,forms, and signaling strategies

The neuregulins (NRGs) are cell-cell signaling proteins that are ligands for receptor tyrosine kinases of the ErbB family. The neuregulin family of genes has four members: NRG1, NRG2, NRG3, and NRG4. Relatively little is known about the biological functions of the NRG2, 3, and 4 proteins, and they are considered in this review only briefly. The NRG1 proteins play essential roles in the nervous system, heart, and breast. There is also evidence for involvement of NRG signaling in the development and function of several other organ systems, and in human disease, including the pathogenesis of schizophrenia and breast cancer. There are many NRG1 isoforms, raising the question "Why so many neuregulins?" Study of mice with targeted mutations ("knockout mice") has demonstrated that isoforms differing in their N-terminal region or in their epidermal growth factor (EGF)-like domain differ in their in vivo functions. These differences in function might arise because of differences in expression pattern or might reflect differences in intrinsic biological characteristics. While differences in expression pattern certainly contribute to the observed differences in in vivo functions, there are also marked differences in intrinsic characteristics that may tailor isoforms for specific signaling requirements, a theme that will be emphasized in this review.     

Reactive oxygen species signaling and stomatal movement in plant responses to drought stress and pathogen attack

Stomata, the pores formed by a pair of guard cells, are the main gateways for water transpiration and photosynthetic CO_2 exchange, as well as pathogen invasion in land plants. Guard cell movement is regulated by a combination of environmental factors, including water status, light, CO_2 levels and pathogen attack, as well as endogenous signals, such as abscisic acid and apoplastic reactive oxygen species(ROS). Under abiotic and bioticstress conditions, extracellular ROS are mainly produced by plasma membrane-localized NADPH oxidases, whereas intracellular ROS are produced in multiple organelles. These ROS form a sophisticated cellular signaling network, with the accumulation of apoplastic ROS an early hallmark of stomatal movement. Here, we review recent progress in understanding the molecular mechanisms of the ROS signaling network,primarily during drought stress and pathogen attack. We summarize the roles of apoplastic ROS in regulating stomatal movement, ABA and CO_2 signaling, and immunity responses.Finally, we discuss ROS accumulation and communication between organelles and cells. This information provides a conceptual framework for understanding how ROS signaling is integrated with various signaling pathways during plant responses to abiotic and biotic stress stimuli.     

Maize cystatins respond to developmental cues, cold stress and drought

Comprehensive searches of maize EST data allowed us to identify 8 novel Corn Cystatin (CC) genes in addition to the previously known genes CCI and CCII. The deduced amino acid sequences of all 10 genes contain the typical cystatin family signature. In addition, they show an extended overall similarity with cystatins from other species that belong to several different phyto-cystatin subfamilies. To gain further insight into their respective roles in the maize plant, gene-specific expression profiles were established by semi-quantitative RT-PCR. While 7 CC genes were expressed in two or more tissues varying from gene to gene, CCI was preferentially expressed in immature tassels and CC8 and CC10 in developing kernels. As shown by in situ hybridisation of maize kernels, CC8 was specifically expressed in the basal region of the endosperm and CC10 both in the starchy endosperm and the scutellum of the embryo. The remaining, not kernel-specific genes, all had distinct expression kinetics during kernel development, generally with peaks during the early stages. In addition to developmental regulation, the effect of cold stress and water starvation were tested on cystatin expression. Two genes (CC8 and CC9) were induced by cold stress and 5 genes (CCII, CC3, CC4, CC5 and CC9) were down-regulated in response to water starvation. Taken together our data suggest distinct functions for CC genes in the maize plant.     

Photosynthetic response of four fern species from different habitats to drought stress: relationship between morpho-anatomical and physiological traits

Ferns flourish in many habitats, from epiphytic to terrestrial and from sunny to shady, and such varied conditions require contrasting photosynthetic strategies to cope with drought. Four species of temperate ferns from different habitats were subjected to drought by withholding irrigation in order to investigate their photosynthetic responses. Lepisorus thunbergianus (epiphytic) had low stomatal density and showed high water-use efficiency (WUE) retaining photosynthetic activity with low relative frond water content under drought stress, which suggested their high adaptation to drought. On the other hand, low WUE with low light-saturated photosynthetic rate in Adiantum pedatum (terrestrial, shady environment) was associated with much lower photosynthesis than in the other species under drought stress, suggesting lower adaptation to drought-prone habitats. Morphological stomatal traits such as stomatal density and photosynthetic response to drought in ferns involved species-specific adaptation to survive and grow in their natural habitats with different levels of drought.     

Rapid responses of C14 clone of Eucalyptus globulus to root drought stress: Time-course of hormonal and physiological signaling

The responses of juvenile plants of forest crops to drought stress are a key stage in the survival of forest populations. In this work, a suitable experimental system to study the early drought resistance mechanisms and signaling in a drought-tolerant clone (C14) of Eucalyptus globulus Labill is proposed. This system, using hydroponic culture and an osmotic agent, polyethylene glycol 8000, was demonstrated to induce severe stress in the root area, affecting the responses of the plantlets at the aerial level. These responses were very fast, beginning only 3 h after the induction of stress, and the results highlight the roles of xylematic abscisic acid (ABA) and pH changes over other signals, such as cytokinins, as early chemical signals in rapid water stress. The relationship between these chemical factors, ABA and pH, and the physiological and water parameters observed were significant, supporting their proposed principal role. This work aids our understanding of underlying responses to hydrological limitations of forest crops, and provides valuable information for further physiological and molecular studies of water stress in this and other tree species.     

Whole plant responses, key processes, and adaptation to drought stress: the case of rice

Most high-yielding rice cultivars developed for irrigated conditions, including the widely grown lowland variety IR64, are highly susceptible to drought stress. This limits their adoption in rainfed rice environments where there is a risk of water shortage during the growing season. Mapping studies using lowland-by-upland rice populations have provided limited information about the genetic basis of variation in yield under drought. One approach to simultaneously improve and understand rice drought tolerance is to generate backcross populations, select superior lines in managed stress environments, and then evaluate which features of the selected lines differ from the recurrent parent. This approach was been taken with IR64, using a range of tolerant and susceptible cultivars as donor parents. Yields of the selected lines measured across 13 widely contracting water environments were generally greater than IR64, but genotype-by-environment effects were large. Traits expected to vary between IR64 and selected lines are plant height, because many donors were not semi-dwarf types, and maturity, because selection in a terminal stress environment is expected to favour earliness. In these experiments it was found that some lines that performed better under upland drought were indeed taller than IR64, but that shorter lines with good yield under drought could also be identified. In trials where drought stress developed in previously flooded (lowland) fields, height was not associated with performance. There was little change in maturity with selection. Other notable differences between IR64 and the selected backcross lines were in their responses to applied ABA and ethylene in greenhouse experiments at the vegetative stage and in leaf rolling observed under chronic upland stress in the field. These observations are consistent with the hypothesis that adaptive responses to drought can effectively allow for improved performance across a broad range of water environments. The results indicate that the yield of IR64 under drought can be significantly improved by backcrossing with selection under stress. In target environments where drought is infrequent but significant in certain years, improved IR64 with greater drought tolerance would be a valuable option for farmers.     

马尾松菌根化苗木对干旱的生理响应及抗旱性评价

采用温室盆栽方法,研究了持续干旱及复水处理后,接种褐环乳牛肝菌7、牛肝菌1、鸡油菌、彩色豆马勃和土生空团菌的马尾松苗木生理变化,并对菌根化苗木进行抗旱性评价.结果表明:在持续干旱条件下,马尾松苗木的丙二醛和相对质膜透性随之增加,但菌根化苗木的丙二醛和相对质膜透性均显著低于未接种苗木(对照);复水后,菌根化苗木中丙二醛和质膜透性较对照迅速降低.在持续干旱胁迫前21 d,马尾松苗木超氧阴离子自由基产生速率增加,同时也诱导了菌根化苗木中过氧化物歧化酶、过氧化物酶和硝酸还原酶活性显著增加.随着胁迫时间的延长,苗木复水后的恢复能力各异.在胁迫14 d复水后,苗木过氧化物歧化酶、过氧化物酶和硝酸还原酶的活性均得以恢复.菌根化苗木抗旱性的强弱为牛肝菌7＞牛肝菌1＞鸡油菌＞土生空团菌＞彩色豆马勃.过氧化物歧化酶和丙二醛与马尾松菌根化苗木抗旱性关联度较大,可以作为抗旱鉴定的主要指标.     

Synthetic and semi-synthetic strategies to study ubiquitin signaling

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(h)GR,beans and drought stress

Reactive oxygen species (ROS) are commonly found in plants as natural by-products of the metabolism but their production is greatly enhanced under abiotic stresses. Particular metabolites and enzymes belonging to the ascorbate-glutathione cycle are able to scavenge these deleterious molecules and modulate the cellular redox-status. In the March issue of Journal of Plant Physiology, we have shown that drought stress induces a raise in glutathione reductase (GR) activity and gene expression that could be related to the intensity of the drought treatment and the drought susceptibility of the bean cultivar (cowpea and/or common bean). In the present addendum we show new data on GR specific activity during progressive drought stress and recovery of the drought-susceptible bean cultivar which can be related to the previously found dual-targeted GR gene expression. Furthermore, since in leguminous plants homoglutathione (hGSH) is generally the most abundant low molecular weight thiol form, we discuss on the occurrence of a (homo)glutathione reductase activity in beans.Key words: common bean, cowpea, drought stress, (homo)glutathione, (homo)glutathione reductase, legumes, Phaseolus vulgaris, recovery, Vigna unguiculataDrought stress is the most common form of abiotic stress and plants are likely to encounter periods of water shortage at least once in their lifecycle. One of the inevitable consequences of drought stress is enhanced reactive oxygen species (ROS) production which will imbalance the cellular redox-status. This shift in the steady-state cellular redox-status is currently believed to have an initial signaling effect, triggering adaptive/defense responses (reviewed in ref. ¹). However, in order to avoid oxidative stress, enhanced ROS production must be kept under tight control by the cellular antioxidant machinery. Glutathione reductase (GR; EC 1.6.4.2) is a major cellular antioxidant enzyme. It belongs to the ascorbate-glutathione cycle and it is ubiquitously found in all cellular compartments.² Using several bean plants (common bean, Phaseolus vulgaris and cowpea, Vigna unguiculata) as a model system to study drought responses and relate them to the degree of drought tolerance and/or susceptibility, we have shown that severe drought stress leads to an enhanced cellular GR activity related to the drought susceptibility of the cultivar.^3,4 Similar results have also been found in a wheat system composed of drought-tolerant and susceptible cultivars.⁵ Regarding the more susceptible cultivar of our bean system (P.v. Carioca) and under severe drought stress (S3, Ψ_w = −2.0 MPa; RWC = 50.9%), total leaf GR activity was raised to approximately 200% when compared to control plants (C, Ψ_w = −0.5 MPa; RWC = 95.3%) (Fig. 1). This could translate a higher degree of oxidative stress due to enhanced ROS production in drought-susceptible cultivars than in drought-tolerant ones. In fact it has been shown that at the cellular level these drought-susceptible bean plants suffer a higher degree of membrane integrity loss when compared to the drought-tolerant.^6–8 This can be related to enhanced ROS production since proteins and lipids of cellular membranes are main targets of ROS peroxidation.⁹Open in a separate windowFigure 1GR-specific activity and relative water content (RWC%) in common bean (Phaseolus vulgaris) ‘Carioca’ leaves. GR-specific activity and RWC were measured in control, severely drought stressed and on rewatered plants. Values are means ± s.d. of three to five independent measurements. GR activity was assayed by following the oxidation of NADPH (decrease in absorbance at 340 nm) and expressed in nmoles min⁻¹ mg⁻¹ protein. RWC was measured according to Weatherley.²⁴ Control plants (C), Ψ_w = −0.5 MPa; severely droughted plants (S3), Ψ_w = −2.0 MPa; 24 h rehydrated plant (24R), Ψ_w = −0.5 MPa; 48 h rehydrated plant (48R), Ψ_w = −0.5 MPa.Considering the responses to drought at the whole-plant level, susceptible and tolerant beans also differ. In fact, drought-tolerant bean cultivars present a water-saving strategy by precocious control of stomatal opening which allows for photosynthetic activity to proceed at lower leaf Ψ_w.^10–12 The maintenance of stomatal opening and photosynthetic activity during drought stress results in lower ROS production by photorespiration and/or the Mehler reaction as opposed to complete stomatal closure where inhibition of CO₂ fixation occurs.¹ Indeed, in the drought-tolerant cowpea cultivar, total GR activity was found constant throughout the progressive drought treatment.³After 24 h rewatering (24R, Ψ_w = −0.5 MPa; RWC = 88.1%), from a moderate water stress: Ψ_w = −1.5 MPa; RWC = 69.2%, GR activity in the drought-susceptible bean cultivar was further raised by ∼270% as compared to control (Fig. 1). This enhanced GR activity can be directly related to the upregulation of the dual-targeted form of the bean GR gene (PvGRdt) (targeted to both chloroplasts and mitochondria) observed on rewatering of this drought-susceptible cultivar.⁴ In fact a significant upregulation of PvGRdt was detected as soon as 6 h after rewatering and the high expression levels were maintained up to 24 h after rewatering to then decrease at 48 h after rewatering.⁴ Hence in the drought-susceptible cultivar it seems that the dual-targeted form is more responsive to drought and rewatering than the cytosolic form. The same pattern was also seen on the less tolerant cowpea cultivar.³ Enhanced dual-targeted GR expression (and GR activity) under drought could be related to enhanced ROS production at those particular cellular compartments (mitochondria and chloroplasts). In fact, under a PEG-induced water deficit, drought-susceptible bean plants showed a higher number of disorganized chloroplasts when compared to the drought-tolerant,⁷ indicating that these organelles experienced oxidative stress during the treatment.The GR enzyme is responsible for the reduction of glutathione disulfide (GSSG) to glutathione (GSH) using NADPH, and not only it keeps glutathione in the reduced state but it is also responsible for the maintenance of the cellular GSH:GSSG ratio.^13,14 Interestingly, in leguminous plants such as the present bean plants, homoglutathione (hGSH) replaces completely or in part, glutathione (GSH). Homoglutathione has been shown to be the most abundant tripeptide in common bean and pea (Pisum sativum),¹⁵ in soybean (Glycine max),¹⁶ and in Lotus japonicus.¹⁷ The synthesis of hGSH proceeds through two ATP-dependent steps, the first step being common with GSH synthesis, the second step adding a β-Alanine instead of a Glycine to form the tripeptide. In the case of cowpea, which was up to now considered to be a non hGSH producing legume,¹⁵ we have recently detected the presence of a homoglutathione synthetase (hGSHS) mRNA and activity (MH Cruz de Carvalho, J Brunet, A Lameta, Y Zuily-Fodil and D Contour-Ansel, unpublished data).Besides the chemical difference of the two thiol tripeptides, many of the roles ascribed to GSH are also performed by hGSH,^18,19 particularly the control of the cellular redox status and ROS scavenging.²⁰ However, the presence of hGSH questions on the occurrence of a homoglutathione reductase (hGR). A role for hGR as a detoxifying enzyme of the ascorbate-glutathione cycle has been suggested, using hGSSG (oxidized homoglutathione) instead of GSSG (oxidized glutathione), thus maintaining the cellular homoglutathione pool in the reduced state and acting as an antioxidative molecule in these plants.^21,22 It can hence be suggested that in beans and other leguminous plants where both thiols co-exist (hGSH and GSH), the (h)GR enzyme will act as either a GR or a hGR in accordance to the thiol utilized in the ascorbate-glutathione cycle.     

Diabetes,oxidative stress and therapeutic strategies

Background

Diabetes has emerged as a major threat to health worldwide.

Scope of Review

The exact mechanisms underlying the disease are unknown; however, there is growing evidence that excess generation of reactive oxygen species (ROS), largely due to hyperglycemia, causes oxidative stress in a variety of tissues. Oxidative stress results from either an increase in free radical production, or a decrease in endogenous antioxidant defenses, or both. ROS and reactive nitrogen species (RNS) are products of cellular metabolism and are well recognized for their dual role as both deleterious and beneficial species. In type 2 diabetic patients, oxidative stress is closely associated with chronic inflammation. Multiple signaling pathways contribute to the adverse effects of glucotoxicity on cellular functions. There are many endogenous factors (antioxidants, vitamins, antioxidant enzymes, metal ion chelators) that can serve as endogenous modulators of the production and action of ROS. Clinical trials that investigated the effect of antioxidant vitamins on the progression of diabetic complications gave negative or inconclusive results. This lack of efficacy might also result from the fact that they were administered at a time when irreversible alterations in the redox status are already under way. Another strategy to modulate oxidative stress is to exploit the pleiotropic properties of drugs directed primarily at other targets and thus acting as indirect antioxidants.

Major Conclusions

It appears important to develop new compounds that target key vascular ROS producing enzymes and mimic endogenous antioxidants.

General significance

This strategy might prove clinically relevant in preventing the development and/or retarding the progression of diabetes associated with vascular diseases.     

Changes in response to drought stress of triticale and maize genotypes differing in drought tolerance

Direct effects and after-effects of soil drought for 7 and 14 d were examined on seedling dry matter, leaf water potential (ψ), leaf injury index (LI), and chlorophyll (Chl) content of drought (D) resistant and sensitive triticale and maize genotypes. D caused higher decrease in number of developed leaves and dry matter of shoots and roots in the sensitive genotypes than in the resistant ones. Soil D caused lower decrease of ψ in the triticale than maize leaves. Influence of D on the Chl b content was considerably lower than on the Chl a content. In triticale the most harmful D impact was observed for physiologically younger leaves, in maize for the older ones. A period of 7-d-long recovery was too short for a complete removal of an adverse influence of D.     

Oxidative stress, insulin signaling, and diabetes

Oxidative stress has been implicated as a contributor to both the onset and the progression of diabetes and its associated complications. Some of the consequences of an oxidative environment are the development of insulin resistance, β-cell dysfunction, impaired glucose tolerance, and mitochondrial dysfunction, which can lead ultimately to the diabetic disease state. Experimental and clinical data suggest an inverse association between insulin sensitivity and ROS levels. Oxidative stress can arise from a number of different sources, whether disease state or lifestyle, including episodes of ketosis, sleep restriction, and excessive nutrient intake. Oxidative stress activates a series of stress pathways involving a family of serine/threonine kinases, which in turn have a negative effect on insulin signaling. More experimental evidence is needed to pinpoint the mechanisms contributing to insulin resistance in both type 1 diabetics and nondiabetic individuals. Oxidative stress can be reduced by controlling hyperglycemia and calorie intake. Overall, this review outlines various mechanisms that lead to the development of oxidative stress. Intervention and therapy that alter or disrupt these mechanisms may serve to reduce the risk of insulin resistance and the development of diabetes.