Efeito do estresse hídrico e envelhecimento precoce na viabilidade de sementes osmocondicionadas de paineira (Chorisia speciosa)

O objetivo deste trabalho foi avaliar a eficiência do condicionamento osmótico em aumentar a tolerância de sementes de paineira (Chorisia speciosa St. Hil.) ao estresse hídrico e ao envelhecimento precoce. As sementes foram condicionadas em solução de KNO3 (0,1M) a 20ºC durante 24 horas, e desidratadas a 27ºC até atingirem o peso inicial, apresentado antes do tratamento. Em seguida, foram expostas ao estresse hídrico simulado com soluções de PEG 6000 nos potenciais osmóticos de 0,0, -0,1, -0,2, -0,3, -0,4, -0,5, -0,6 e -0,7 MPa e aos efeitos do envelhecimento precoce (45ºC e 100% UR) durante 0, 24, 48, 72, 96 e 120 horas. Nas sementes condicionadas e submetidas ao estresse hídrico, os valores médios de porcentagem e velocidade de germinação foram estatisticamente superiores aos obtidos em sementes não condicionadas. Além disso, o condicionamento osmótico aumentou o limite máximo de tolerância ao estresse hídrico. As sementes condicionadas foram mais sensíveis ao envelhecimento precoce.

Comportamento de linhagens de sorgo forrageiro submetidas a déficit hídrico sob condição controlada

O objetivo deste trabalho foi avaliar os efeitos do estresse hídrico no estabelecimento, desenvolvimento e na taxa de sobrevivência de 11 linhagens de sorgo forrageiro em casa de vegetação. O delineamento experimental foi o de blocos ao acaso no arranjo de parcelas subdivididas no tempo, com cinco repetições. As unidades experimentais foram constituídas por recipientes de 250 L, onde foram colocados 187,9 kg de solo da classe textural areia franca. O estudo compreendeu dois períodos de estresse, e três irrigações, sendo uma no início e as outras aplicadas quando grande número de folhas tinham aspecto de palha seca. No primeiro período, as linhagens apresentaram alta taxa de sobrevivência, demonstrando resistência ao estresse hídrico. As linhagens que apresentaram menor velocidade de crescimento nesse período sofreram menos com o estresse. No segundo período não ocorreram diferenças significativas entre as linhagens com relação ao desenvolvimento das plantas. Durante os dois períodos de estresse, a velocidade de crescimento não diferenciou satisfatoriamente linhagens de comportamento distintos, quanto à resistência à seca, competindo em um mesmo recipiente e em alta densidade populacional. As linhagens 84, 29, 99, 41, 63 e 104 apresentaram maior resistência ao estresse hídrico e as linhagens 33, 38, 40, 67 e 92, menor resistência.

Condicionamento osmótico na germinação de sementes de cássia-do-nordeste sob estresse hídrico, térmico e salino

O objetivo deste trabalho foi avaliar o efeito do condicionamento osmótico na germinação de sementes de Cassia excelsa Schrad. sob condições de estresse hídrico, térmico e salino. O condicionamento consistiu na imersão de quatro repetições de 25 sementes cada, em soluções de polietilenoglicol (PEG 6000) com potenciais osmóticos de -0,2, -0,4 e -0,6 MPa por 72, 96, 168 horas, respectivamente, e em água destilada por 48 horas, a 20ºC. Em seguida, as sementes foram colocadas para germinar em três condições: estresse hídrico, simulado com soluções de PEG a -0,2, -0,4, -0,6 e -0,8 MPa a 27ºC; estresse térmico, sob temperaturas subótima (12ºC) e supra-ótima (39ºC) em água destilada; estresse salino, em soluções de NaCl a -0,2, 0,4, 0,6, 0,8, 1,0, 1,2 e 1,4 MPa a 27ºC. Foram avaliadas as porcentagens de germinação e tempos médios de emergência da radícula. A técnica de condicionamento osmótico com PEG ou com água destilada foi eficiente em aumentar a germinabilidade sob estresse hídrico e sob estresse térmico. A porcentagem de germinação sob estresse salino aumentou com o condicionamento com PEG, porém não houve redução no tempo de germinação.

Compactação do solo e intervalo hídrico ótimo na produtividade de arroz de sequeiro

O objetivo deste trabalho foi determinar o intervalo hídrico ótimo e sua relação com a produtividade de grãos de arroz de sequeiro em um Latossolo Vermelho de textura média. Os tratamentos foram 0, 1, 2, 4 e 6 passadas de um trator, lado a lado, na superfície do solo, com quatro repetições. O delineamento experimental foi inteiramente casualizado. Foram coletadas amostras indeformadas de solo nas camadas de 0,02-0,05, 0,07-0,10 e 0,15-0,18 m para determinação da resistência do solo à penetração, densidade do solo, intervalo hídrico ótimo, densidade do solo crítica e da curva de retenção de água no solo. Em parcelas de 3,6 m² foi semeado arroz de sequeiro (cv. Caiapó) e a sua produtividade foi avaliada em parcelas úteis de 1,80 m². O intervalo hídrico ótimo foi reduzido a partir da densidade do solo de 1,30 Mg m-3 pela resistência do solo à penetração. A partir do valor de 1,82 MPa ocorreu decréscimo da produtividade de arroz. A densidade do solo crítica ao desenvolvimento radicular no intervalo hídrico ótimo foi de 1,63 Mg m-3, equivalente a 1,62 Mg m-3 que foi limitante na produtividade de arroz. O monitoramento da compactação do solo é necessário para prevenir perdas na produtividade de arroz de sequeiro.

Processo germinativo de sementes de paineira sob estresses hídrico e salino

O ambiente mutável provoca múltiplos estresses, que podem limitar a germinação da semente e a emergência e sobrevivência da plântula. Os objetivos deste trabalho foram determinar o efeito dos estresses hídrico e salino na viabilidade e vigor de sementes de paineira (Chorisia speciosa St. Hil.) e o limite máximo de tolerância a esses estresses. Para avaliação do efeito do estresse hídrico foram utilizadas soluções de manitol e PEG 6000; para o estresse salino, soluções de NaCl, KCl e CaCl2. Após a punção do tegumento, quatro repetições de 50 sementes por tratamento foram submetidas à germinação em substrato de papel-filtro umedecido com as soluções de diferentes potenciais osmóticos e incubadas a 27ºC. As sementes apresentaram menor tolerância ao estresse hídrico simulado com PEG 6000 em relação ao simulado com manitol. O limite máximo de tolerância à seca está situado entre -0,6 e -0,7 MPa de PEG 6000 e entre -1,4 e -1,6 MPa de manitol. Independentemente dos sais utilizados, a porcentagem de germinação decresceu com a diminuição do potencial osmótico do meio. O limite máximo de tolerância ao estresse salino foi o mesmo para todos os sais avaliados e está situado entre -1,0 e -1,2 MPa. A paineira pode ser classificada como glicófita, com moderada tolerância aos sais NaCl, KCl e CaCl2.

Role of glutathione deficit in the disconnectivity syndrome in schizophrenia: from pathogenetic mechanisms to therapeutic interventions

In the cerebrospinal fluid of 26 drug-naive schizophrenics (DSM-III- R), we observed that the level of glutathione ([GSH]) and of its metabolite γ-Glu-Gln was decreased by 27% and 16% respectively. Using a new in-vivo method based on magnetic resonance spec- troscopy, [GSH] was measured in the medial prefrontal cortex of 18 schizophrenics and found to be 52 % lower than in controls (n = 20). This is consistent with the recently observed decreased mRNA levels in fibroblasts of patients (n=32) of the two GSH synthesizing en- zymes (glutathione synthetase (GSS), and glutamate-cysteine ligase M (GCLM) the modulatory subunit of glutamate-cysteine ligase). Moreover, the level of GCLM expression in fibroblasts correlates neg- atively with the psychopathology (positive, general and some nega- tive symptoms). Thus, the observed difference in gene expression is not only the cause of low brain [GSH], but is also related to the sever- ity of symptoms, suggesting that fibroblasts are adequate surrogate for brain tissue. A hypothesis was proposed, based on a central role of GSH in the pathophysiology of schizophrenia. GSH is an important endogenous redox regulator and neuroactive substance. GSH is pro- tecting cells from damage by reactive oxygen species generated, among others, by the metabolism of dopamine. A GSH deficit-in- duced oxidative stress would lead to lipid peroxidation and micro-le- sions in the surrounding of catecholamine terminals, affecting the synaptic contacts on dendritic spines of cortical neurones, where ex- citatory glutamatergic terminals converge with dopaminergic ones. This would lead to spines degeneration and abnormal nervous con- nections or structural disconnectivity, possibly responsible for posi- tive, perceptive and cognitive symptoms of schizophrenia. In addi- tion, a GSH deficit could also lead to a functional disconnectivity by depressing NMDA neurotransmission, in analogy to phencyclidine effects. Present experimental biochemical, cell biological and behav- ioral data are consistent with the proposed mechanism: decreasing pharmacologically [GSH] in experimental models, with or without blocking DA uptake (GBR12909), induces morphological and behav- ioral changes similar to those observed in patients. Dendritic spines: (a) In neuronal cultures, low [GSH] and DA induce decreased density of neural processes; (b) In developing rats (p5-p16), [GSH] deficit and GBR induce a decrease in normal spines in prefrontal pyramids and in GABA-parvalbumine but not of -calretinine immunoreactivity in anterior cingulate. NMDA-dependant synaptic plasticity: GSH deple- I/13 tion in hippocampal slices impairs long-term potentiation. Develop- ing rats with low [GSH] and GBR have deficit in olfactory integration and in object recognition which appears earlier in males than fe- males, in analogy to the delay of the psychosis onset between man and woman. In summary, a deficit of GSH and/or GSH-related enzymes during early development could constitute a major vulnerability fac- tor in schizophrenia.

Mice with chronic GSH deficit display phenotypical anomalies that resemble key aspects of schizophrenia

ABSTRACTSchizophrenia is a major psychiatric disorder occurring with a prevalence of 1% in the worldwide population. It develops progressively with psychosis onset in late adolescence or earlyadulthood. The disorder can take many different facets and has a highly diffuse anddistributed neuropathology including deficits in major neurotransmitter systems,myelination, stress regulation, and metabolism. The delayed onset and the heterogeneouspathology suggest that schizophrenia is a developmental disease that arises from interplayof genetic and environmental factors during sensitive periods. Redox dysregulation due to animbalance between pro-oxidants and antioxidant defence mechanisms is among the riskfactors for schizophrenia. Glutathione (GSH) is the major cellular redox regulator andantioxidant. Levels of GSH are decreased in cerebrospinal fluid, prefrontal cortex and postmortemstriatum of schizophrenia patients. Moreover, polymorphisms of the key GSHsynthesizingenzyme, glutamate-cysteine ligase, modifier (GCLM) subunit, are associatedwith the disease, suggesting that GSH deficit is of genetic origin. Here we used miceknockout (KO) for the GCLM gene, which display chronic GSH deficit (~70 to 80% decrease)to investigate the direct link between redox dysregulation and schizophrenia. Accordingly,we evaluated whether GCLM KO compared to normal wildtype mice display behavioralchanges that relate to schizophrenia symptoms and whether their brains showmorphological, functional or metabolic alterations that resemble those in patients.Moreover, we exposed pubertal GCLM mice to repeated mild stress and measured theirhormonal and behavioral stress reactivity. Our data show that chronic GSH deficit isassociated with altered emotion- and stress-related behaviors, deficient prepulse inhibition,pronounced amphetamine-induced hyperlocomotion but normal spatial learning andworking memory. These changes represent important schizophrenia endophenotypes.Moreover, this particular pattern of change indicates impairment of the ventralhippocampus (VH) and related circuitry as opposed to the dorsal hippocampus (DH), which isimplicated in spatial information processing. This is consistent with a selective deficit ofparvalbumin positive interneurons and gamma oscillation in the VH but not DH. Increasedlevels of circulating stress hormones in KO mice following pubertal stress corroborate VHdysfunction as it is involved in negative feedback control of the stress response. VHstructural and functional deficits are frequently found in the schizophrenic brain. Metabolicevaluation of the developing GCLM KO anterior cortex using in vivo magnetic resonancespectroscopy revealed elevated glutamine (Gln), glutamate (Glu), Gln/Glu and N-acetylaspartate(NAA) during the pre-pubertal period. Similar changes are reported in earlyschizophrenia. Overall, we observe phenotypic anomalies in GSH deficient GCLM KO micethat correspond to major schizophrenia endophenotypes. This supports an important rolefor redox dysregulation in schizophrenia and validates the GCLM KO mouse as model for thedisease. Moreover, our results indicate that puberty may be a sensitive period for redoxsensitivechanges highliting the importance of early intervention. Gln, Gln/Glu, Glu and NAAmay qualify as early metabolic biomarkers to identify young at-risk individuals. Since chronictreatment with NAC normalized most metabolic changes in GCLM KO mice, NAC may be oneadjunct treatment of choice for early intervention in patients.RESUMELa schizophrénie est une maladie psychiatrique majeure avec une prévalence de 1% dans lapopulation. Son développement est progressif, les premières psychoses apparaissant àl'adolescence ou au début de l'âge adulte. La maladie a plusieurs présentations et uneneuropathologie étendue, qui inclut des déficits neurochimiques, métaboliques, de lamyélination et de la régulation du stress. L'émergence tardive et l'hétérogénéité de lapathologie suggèrent que la schizophrénie est une maladie développementale, favorisée pardes facteurs génétiques et environnementaux durant des périodes sensibles. La dérégulationrédox, due à un déséquilibre entre facteurs pro-oxidantes et défenses anti-oxidantes,constitue un facteur de risque. Le glutathion (GSH) est le principal régulateur rédox et antioxidantdes cellules, ses taux sont diminués dans le liquide céphalorachidien, le cortexpréfrontal et le striatum de patients. De plus, des variations du gène codant la sous-unitémodulatrice (GCLM) de la glutamate-cystéine ligase, enzyme de synthèse du GSH, sontassociés la maladie, suggérant que le déficit observé chez les patients est d'originegénétique. Nous avons donc utilisé des souris ayant une délétion du gène GCLM (KO), quiont un déficit chronique en GSH (70-80%), afin d'étudier le lien entre une dérégulation rédoxet la schizophrénie. Nous avons évalué si ces souris présentent des altérationscomportementales analogues aux symptômes de la maladie, et des modificationsstructurelles, fonctionnelles et métaboliques au niveau du cerveau, ressemblant à celles despatients. De plus, nous avons soumis les souris à des stresses modérés durant la puberté,puis mesuré les réponses hormonales et comportementales. Les animaux présentent undéficit pré-attentionnel du traitement des informations moto-sensorielles, un déficit pourcertains apprentissages, une réponse accrue à l'amphétamine, mais leurs mémoires spatialeet de travail sont préservées. Ces atteintes comportementales sont analogues à certainsendophénotypes de la schizophrénie. De plus, ces changements comportementaux sontlargement expliqués par une perturbation morphologique et fonctionnelle de l'hippocampeventral (HV). Ainsi, nous avons observé un déficit sélectif des interneurones immunoréactifsà la parvalbumine et une désynchronisation neuronale dans l'HV. L'hippocampe dorsal,impliqué dans l'orientation spatiale, demeure en revanche intact. L'augmentationd'hormones de stress dans le sang des souris KO suite à un stress prépubertal soutien aussil'hypothèse d'une dysfonction de l'HV, connu pour moduler ce type de réponse. Des déficitsstructurels et fonctionnels dans l'hippocampe antérieur (ventral) ont d'ailleurs été rapportéschez des patients schizophrènes. Par de résonance magnétique, nous avons également suivile profil métabolique du le cortex antérieur au cours du développement postnatal des sourisKO. Ces mesures ont révélé des taux élevés de glutamine (Gln), glutamate (Glu), du ratioGln/Glu, et de N-acétyl-aspartate (NAA) durant la période prépubertale. Des altérationssimilaires sont décrites chez les patients durant la phase précoce. Nous avons donc révélédes anomalies phénotypiques chez les souris GCLM KO qui reflètent certainsendophénotypes de la schizophrénie. Nos résultats appuient donc le rôle d'une dérégulationrédox dans l'émergence de la maladie et le potentiel des souris KO comme modèle. De plus,cette étude met en évidence la puberté comme période particulièrement sensible à unedérégulation rédox, renforçant l'importance d'une intervention thérapeutique précoce. Dansce cadre, Gln, Gln/Glu, Glu and NAA seraient des biomarqueurs clés pour identifier de jeunesindividus à risque. De part son efficacité dans notre modèle, NAC pourrait être unesubstance de choix dans le traitement précoce des patients.

Early glutathione deficit impairs parvalbumin expression in gaba interneurons and kainate-induced gamma oscillations

An increased oxidative stress and alteration of the antioxidant systems have been observed in schizophrenia. Glutathione (GSH), a major redox regulator, is decreased in patients' cerebrospinal fluid, prefrontal cortex in vivo and striatum post-mortem tissue. Most importantly, there is genetic and functional evidence for the implication of the gene of the glutamate cysteine ligase (GCL) catalytic subunit, the key GSH-synthesizing enzyme. We have developed animal models for a GSH deficit to study the consequences of such deficit on the brain development. A GSH deficit combined with elevated dopamine (DA) during development leads to reduced parvalbumin (PV) expression in a subclass of GABA interneurons in rat anterior cingulate cortex (ACC). Similar changes are observed in postmortem brain tissue of schizophrenic patients. GSH dysregulation increases vulnerability to oxidative stress, that in turn could lead to cortical circuit anomalies in the schizophrenic brain. In the present study, we use a GCL modulatory subunit (GCLM) knock-out (KO) mouse model that presents up to 80% decreased brain GSH levels. During postnatal development, a subgroup of animals from each genotype is exposed to elevated oxidative stress induced by treatment with the DA reuptake inhibitor GBR12909. Results reveal a significant genotype-specific delay International Congress on Schizophrenia Research 136 10. 10. Neuroanatomy, Animal Downloaded from http://schizophreniabulletin.oxfordjournals.org at Bibliotheque Cantonale et Universitaire on June 18, 2010 in cortical PV expression at postnatal day P10 in GCLM-KO mice, as compared to wild-type. This effect seems to be further exaggerated in animals treated with GBR12909 from P5 to P10. At P20, PV expression is no longer significantly reduced in GCLM-KO ACC without GBR but is reduced if GBR is applied from P10 to P20. However, our result show that GCLM-KO mice exhibit increased oxidative stress, cortical altered myelin development as shown by MBP marker, and more specifically impairment of the peri-neuronal net known to modulate PV connectivity. In addition, we also observe a reduced PV expression in the ventro-temporal hippocampus of adult GCLM-KO mice, suggesting that anomalies of the PV interneurons prevail at least in some brain regions throughout the adulthood. Interestingly, the power of kainate-induced gamma oscillations, known to be dependent on proper activation of PV interneuron's, is also lower in hippocampal slices of adult GCLM KO mice. These results suggest that the PV positive GABA interneurons is particularly vulnerable to increased oxidative stress

Brazilian maize genotypes sensitivity to water deficit estimated through a simple crop yield model

The objective of this work was to determine the sensitivity of maize (Zea mays) genotypes to water deficit, using a simple agrometeorological crop yield model. Crop actual yield and agronomic data of 26 genotypes were obtained from the Maize National Assays carried out in ten locations, in four Brazilian states, from 1998 to 2006. Weather information for each experimental location and period were obtained from the closest weather station. Water deficit sensitivity index (Ky) was determined using the crop yield depletion model. Genotypes can be divided into two groups according to their resistance to water deficit. Normal resistance genotypes had Ky ranging from 0.4 to 0.5 in vegetative period, 1.4 to 1.5 in flowering, 0.3 to 0.6 in fruiting, and 0.1 to 0.3 in maturing period, whereas the higher resistance genotypes had lower values, respectively 0.2-0.4, 0.7-1.2, 0.2-0.4, and 0.1-0.2. The general Ky for the total growing season was 2.15 for sensitive genotypes and 1.56 for the resistant ones. Model performance was acceptable to evaluate crop actual yield, whose average errors estimated for each genotype ranged from -5.7% to +5.8%, and whose general mean absolute error was 960 kg ha-1 (10%).

Metabolic alterations in the cortex of a mouse model with glutathione deficit - relevance to schizophrenia

Background: Glutathione (GSH) is a major redox regulator and antioxidant and is decreased in cerebrospinal fluid and prefrontal cortex of schizophrenia patients [Do et al. (2000) Eur J Neurosci 12:3721]. The genes of the key GSH-synthesizing enzyme, glutamate- cysteine ligase catalytic (GCLC) and modifier (GCLM) subunits, are associated with schizophrenia, suggesting that the deficit in GSH synthesis is of genetic origin [Gysin et al. (2007) PNAS 104:16621]. GCLM knock-out (KO) mice, which display an 80% decrease in brain GSH levels, have abnormal brain morphology and function [Do et al. (2009) Curr Opin Neurobiol 19:220]. Developmental redox deregulation by impaired GSH synthesis and environmental risk factors generating oxidative stress may have a central role in schizophrenia. Here, we used GCLM KO mice to investigate the impact of a genetically dysregulated redox system on the neurochemical profile of the developing brain. Methods: The neurochemical profile of the anterior and posterior cortical areas of male and female GCLM KO and wild-type mice was determined by in vivo 1H NMR spectroscopy on postnatal days 10, 20, 30, 60 and 90, under 1 to 1.5% isoflurane anaesthesia. Localised 1H NMR spectroscopy was performed on a 14.1 T, 26 cm VNMRS spectrometer (Varian, Magnex) using a home-built 8 mm diameter quadrature surface coil (used both for RF excitation and signal reception). Spectra were acquired using SPECIAL with TE of 2.8 ms and TR of 4 s from VOIs placed in anterior or posterior regions of the cortex [Mlynárik et al. (2006) MRM 56:965]. LCModel analysis allowed in vivo quantification of a neurochemical profile composed of 18 metabolites. Results: GCLM KO mice displayed nearly undetectable GSH levels as compared to WT mice, demonstrating their drastic redox deregulation. Depletion of GSH triggered alteration of metabolites related to its synthesis, namely increase of glycine and glutamate levels during development (P20 and P30). Concentrations of glutamine and aspartate that are produced from glutamate were also increased in GCLM KO animals relative to WT. In addition, GCLM KO mice also showed higher levels of N-acetylaspartate that originates from the acetylation of aspartate. These metabolites are particularly implicated in neurotransmission processes and in mitochondrial oxidative metabolism. Their increase may indicate impaired mitochondrial metabolism with concomitant accumulation of lactate in the adult mice (P60 and P90). In addition, the GSH depletion triggers reduction of GABA concentration in anterior cortex of the P60 mice, which is in accordance with known impairment of GABAergic interneurons in that area. Changes were generally more pronounced in males than in females at P60, which is consistent with earlier disease onset in male patients. Discussion: In conclusion, the observed metabolic alterations in the cortex of a mouse model of redox deregulation suggest impaired mitochondrial metabolism and altered neurotransmission. The results also highlight the age between P20 and P30 as a sensitive period during the development for these alterations.

Septal lesions impair the acquisition of a cued place navigation task: attentional or memory deficit?

These experiments were designed to analyze how medial septal lesions reducing the cholinergic innervation in the hippocampus might affect place learning. Rats with quisqualic lesions of the medial septal area (MS) were trained in a water maze and on a homing table where the escape position was located at a spatially fixed position and further indicated by a salient cue suspended above it. The lesioned rats were significantly impaired in reaching the cued escape platform during training. In addition rats, did not show any discrimination of the training sector during a probe trial in which no platform or cue was present. This impairment remained significant during further training in the absence of the cue. When the cued escape platform was located at an unpredictable spatial location, the MS-lesioned rats showed no deficit and spent more time under the cue than control rats during the probe trial. On the homing board, with a salient object in close proximity to the escape hole, the MS rats showed no deficit in escape latencies, although a significant reduction in spatial memory was observed. However, this was overcome by additional training in the absence of the cue. Under these conditions, rats with septal lesions were prone to develop a pure guidance strategy, whereas normal rats combined a guidance strategy with a memory of the escape position relative to more distant landmarks. The presence of a salient cue appeared to decrease attention to environmental landmarks, thus reducing spatial memory. These data confirm the general hypothesis that MS lesions reduce the capacity to rely on a representation of the relation between several landmarks with different salience.