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Drought stress inhibits oat growth and yield. The application of fulvic acid (FA) can improve the drought resistance of oats, but the corresponding molecular mechanism of FA-mediated drought resistance remains unclear. Here, we studied the effects of FA on the drought tolerance of oat leaves through physiological, transcriptomic, and metabolomics analyses, and identified FA-induced genes and metabolites related to drought tolerance. Physiological analysis showed that under drought stress, FA increased the relative water and chlorophyll contents of oat leaves, enhanced the activity of antioxidant enzymes (SOD, POD, PAL, CAT and 4CL), inhibited the accumulation of malondialdehyde (MDA), hydrogen peroxide (H2O2) and dehydroascorbic acid (DHA), reduced the degree of oxidative damage in oat leaves, improved the drought resistance of oats, and promoted the growth of oat plants. Transcriptome and metabolite analyses revealed 652 differentially expressed genes (DEGs) and 571 differentially expressed metabolites (DEMs) in FA-treated oat leaves under drought stress. These DEGs and DEMs are involved in a variety of biological processes, such as phenylspropanoid biosynthesis and glutathione metabolism pathways. Additionally, FA may be involved in regulating the role of DEGs and DEMs in phenylpropanoid biosynthesis and glutathione metabolism under drought stress. In conclusion, our results suggest that FA promotes oat growth under drought stress by attenuating membrane lipid peroxidation and regulating the antioxidant system, phenylpropanoid biosynthesis, and glutathione metabolism pathways in oat leaves. This study provides new insights into the complex mechanisms by which FA improves drought tolerance in crops.
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Stem rust, caused by Puccinia graminis f. sp. avenae (Pga) is a key disease affecting oat production worldwide. Silicon (Si) plays an essential role in enhancing plant resistance against pathogens. However, the scientific evidence of Si-mediated stem rust resistance of oat from the photosynthetic perspective has not been reported. The specific objective of this research was to investigate the effects of Si application on disease inhibition, photosynthetic gas exchange parameters, light response parameters, photosynthetic pigments and chlorophyll fluorescence parameters under Pga infection. Our results illustrated that Si application significantly reduced rust severity while the other parameters like net photosynthetic rate (P n), stomatal conductance (Gs), intercellular CO2 concentration (C i) and transpiration rate (T r) were significantly increased. Si application increased maximum photosynthetic rate (P nmax) and light saturation point (LSP), while reduced the dark respiration rate (Rd) and light compensation point (LCP). The results also indicated that Si application significantly increased the activities of maximum fluorescence (F m), variable fluorescence (F v), maximum quantum yield of photosystem II (F v/F m), photochemical quenching (qP), photosynthetic performance index (PI ABS), actual PSII quantum yield (ΦPSII), electron transfer rate (ETR), the absorbed light energy per unit reaction center (ABS/RC) and the dissipated energy per unit reaction center (DIo/RC), whereas it decreased the minimal fluorescence (F o), non-photochemical quenching (NPQ), the absorbed light energy used for electron transfer per unit reaction center (ETo/RC) and the absorbed light energy used for reduction of QA per unit reaction center (TRo/RC). The contents of chlorophyll a, b and carotenoids were also increased due to the change in the activity of parameters due to Si application as mentioned above. In conclusion, the results of the current study suggests that Si imparts tolerance to the stem rust possibly by the underlying mechanisms of improving gas exchange performance, and efficiency of the photochemical compounds in oat leaves.
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Oat is considered as a moderately salt-tolerant crop that can be used to improve saline and alkaline soils. Previous studies have focused on short-term salt stress exposure, and the molecular mechanisms of salt tolerance in oat have not yet been elucidated. In this study, the salt-tolerant oat cultivar Vao-9 and the salt-sensitive oat cultivar Bai5 were treated with 6 days of 0 and 150 mmol L-1 salt stress (nNaCl:nNa2SO4 = 1:1). Label-Free technology was then used to analyze the differentially expressed proteins in leaves under 0 and 150 mmol L-1 salt stress. The obtained results indicated that total of 2,631 proteins were identified by mass spectrometry in the four samples. The salt-tolerant cultivar Vao-9 mainly enhances its carbohydrate and energy metabolism through the pentose and glucuronate interconversions, and carbon fixation pathways in prokaryotes, thereby reducing the damage caused by salt stress. In addition, the down-regulation of ribosomes expression and the up-regulated expression of HSPs and CRT are all through the regulation of protein synthesis in response to salt stress. However, GABA metabolism presents a different synthesis pattern in Bai5 and Vao-9. The main KEGG function of differential expressed protein (DEP) in Bai5 is classified into protein processing in the endoplasmic reticulum, estrogen signaling pathway, antigen processing and presentation, longevity regulating pathway-multiple species, arginine and proline metabolism, beta-alanine metabolism, vitamin B6 metabolism, salmonella infection, chloroalkane and chloroalkene degradation, and limonene and pinene degradation. Moreover, the main KEGG functions of DEP in Vao-9 are classified as ribosome and carbon fixation pathways in prokaryotes, pentose and glucuronate interconversions, GABA ergic synapse, and taurine and hypotaurine metabolism. The results obtained in this study provide an important basis for further research on the underlying mechanisms of salt response and tolerance in oat and other plant species.
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With the serious shortage of water resources and the development of water-saving agriculture, the application of drip irrigation has been paid more and more attention. But there was lack of oat planting methods suitable for drip irrigation, currently. In order to establish an efficient oat planting method for drip irrigation, a study was conducted at Agriculture and Forestry Sciences of Ulanqab, Inner Mongolia during the season (2019-2020) to evaluate the effect of strip cropping with reducing row spacing and super absorbent polymer on the yield and water use efficiency of oat. To conduct the field trials, a split plot system in three replications was established. Three planting patterns were in the main plots, including conventional cropping with 20 cm equal row spacing (PA), strip cropping with the 15 cm row spacing (PB) and strip cropping with the 10 cm row spacing (PC), and two super absorbent polymer levels were in the subplots, including 22.5 kg ha-2 (Y) and 0 (N). The results showed that, compared with PA, PB and PC both decreased the irrigation volumes by 4.5-18.4 mm, and the irrigation volumes of PB was lower than that of PC. When super absorbent polymers were applied, compared with PA, PB significantly increased grain yield and above-ground biomass, but PC had the opposite effects. The grain yield and above-ground biomass of PB significantly increased by 16.65% and 7.31% on average in two years, respectively. And the increasing of grain yield was attributed by the significant increasing of pike number and kernel number per spike. But when super absorbent polymers were not applied, PB had no significant effects on grain yield and above-ground biomass. PB also had the significant effects on regulating water use of oats weather or not super absorbent polymers were applied, it significantly increased the precipitation ratio by 2.64% (PBY) and 2.13% (PBN) and decreased irrigation ration by 3.32% (PBY) and 5.28% (PBN) on average in two years. Although PB and PC both decreased the total evapotranspiration, but PB increased WUE and PC deceased WUE. The WUE of PB increased by 19.70% (PBY) and 9.87% (PBN) on average in two years. Also PB had the highest economic benefits in all treatments. In conclusion, a drip irrigation oat planting pattern was proposed, which the row spacing is 15 cm, adjusted the equal row spacing planting to 8-row strip planting, with a belt spacing of 30 cm, combined with the application of 22.5 kg ha-2 applying super absorbent polymers. And this oat planting pattern is a viable strategy to improve oat productivity.
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Avena , Agua , Agricultura/métodos , Grano Comestible , PolímerosRESUMEN
To provide theoretical guidance and technical support for oat production in dry farming area, we exa-mined the effects of moisture and humic acid (HA) on the accumulation and distribution of non-structural carbohydrates (NSC) in different organs of oat, as well as the mechanism of sugar metabolism and grain weight formation of oat. Two oat cultivars, 'Mengnong Dayan 1' and 'Neiyan 5', were used as experimental material. HA and clean water (CK) were foliar sprayed under dry framing (without irrigation) and limited irrigation (irrigated twice at jointing and heading stage). The dynamics of NSC components in stems, leaves and panicles, as well as the changes of carbon metabolism-related enzyme activities at different growth stages of oats after flowering were measured. Results showed that the trend of the contents of NSC in stems, leaves and panicles, in both two oat cultivars increased firstly and then decreased with the prolong of post-anthesis time. The contents of NSC in different organs were similar between two oat cultivars. Under irrigation treatment, the fructan content in panicle for Mengnong Dayan 1 of HA was higher than that of CK, with magnitude of enhancement being significantly greater than that corresponding treatment under dry farming. Under irrigation condition, the fructan, invertase activities in leaves and grain weight per panicle for Mengnong Dayan 1 of HA were increased by 27.1%, 30.6% and 55.9% compared with CK, respectively. Further, the increase trend under irrigation was stronger than that under dry farming condition. For Mengnong Dayan 1, the 1000-grain weight and grain weight per panicle were positively correlated with the content of fructan in leaves. In conclusion, the synergistic effect of moisture and humic acid could effectively regulate the accumulation of oat fructans and the activities of carbon metabolic enzymes, consequently promoting the formation of yield.
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Avena , Fructanos , Avena/metabolismo , Carbohidratos , Carbono/metabolismo , Grano Comestible/metabolismo , Fructanos/metabolismo , Sustancias Húmicas , Hojas de la Planta/metabolismo , Triticum , AguaRESUMEN
The effects of foliar application of spermidine (Spd) on the physiological aspects of salt-stressed oat seedlings were studied under greenhouse conditions. At the seedling stage, the salt-sensitive variety, namely, Caoyou 1 and the salt-tolerant variety, namely, Baiyan 2 were treated with 70 and 100 mM of salt, followed by the foliar application of 0.75 mM Spd or distilled water. Results showed that Spd application increased the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX), and reduced the rate of O2 â - production and the accumulation of H2O2 and malondialdehyde (MDA). In addition, it increased the level of zeatin riboside (ZR) and the content of endogenous polyamines. The application of Spd increased the contents of soluble sugar, soluble protein, and free proline and helped maintain the osmotic balance of oat leaves. At the same time, foliar Spd treatment helped in maintaining the ion nutrition balance. Specifically, it reduced the content of Na+ and thereby stabilized the ratio of Na+/K+, Na+/Ca2+, and Na+/Mg2+. The effects of Spd application were more obvious for the salt-sensitive cultivar Caoyou 1 and under the lighter 70 mM salt stress.
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Soil salinity is among the crucial factors that impact on crop productivity, including oat (Avena sativa L.). Herein, we used two distinct oat cultivars with varied salt tolerance levels to unravel adaptive responses to salt stress by metabolomic and transcriptomic characterization. Metabolomic profiling revealed 201 metabolites, including saccharides, amino acids, organic acids, and secondary metabolites. The levels of most saccharides and amino acids were elevated in Baiyan 2 (BY2) as well as in Baiyan 5 (BY5) exposed to salt stress. In the tolerant cultivar BY2 exposed to 150 mM NaCl, concentrations of most of the metabolites increased significantly, with sucrose increased by 38.34-fold, Sophorose increased by 314.15-fold and Isomaltose 2 increased by 25.76-fold. In the sensitive cultivar BY5, the concentrations of most metabolites increased after the plant was exposed to 150 mM NaCl but decreased after the plant was exposed to 300 mM NaCl. Transcriptomic analysis revealed that gene expressions in BY5 were significantly affected under exposure to 300 mM NaCl (34040 genes up-regulated and 14757 genes down-regulated). Assessment of metabolic pathways as well as KEGG enrichment revealed that salt stress interferes with the biosynthesis of two oat cultivars, including capacity expenditure and sugar metabolism. Most of the BY2 genes enhanced energy consumption (for example, glycolysis) and biosynthesis (for instance, starch and sugar metabolism) under salt stress. In contrast, genes in BY5 were found to be down-regulated, leading to the inhibition of energy consumption and biosynthesis, which may also be attributed to salt sensitivity in BY5. In addition, the modified Na+/K+ transporter genes expression is associated with the predominant ionic responses in BY2, which leads low concentration of Na+ and high K+ when exposed to high salt situations. These findings suggest that the varied defensive capacities of these two oat cultivars in response to salt stress are due to their variations in energy-expenditure strategy, synthesis of energy substances and ion transport in roots. Our present study offers a crucial reference for oat cultivation under saline soil.
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Avena , Metaboloma , Tolerancia a la Sal , Transcriptoma , Avena/genética , Avena/fisiología , Regulación de la Expresión Génica de las Plantas , SalinidadRESUMEN
The application of organic amendments to saline-alkaline soil has been recommended as an agricultural strategy to improve crop productivity and soil health. However, there has been limited research on how organic soil amendment strategies affect the health of oats and their associated rhizosphere fungal communities in saline-alkaline conditions. Thus, the objectives of this study were to understand the effects of oat cultivars with contrasting saline-alkaline tolerances and different amendments on plant morphologies, root exudates (soluble sugars and organic acids), and rhizosphere fungal communities in a saline-alkaline environment. Experiments were conducted on a saline-alkaline tolerant cultivar, Baiyan2, and a saline-alkaline sensitive cultivar, Caoyou1, under four different organic amendment strategies: 1. control (no amendment application), 2. bio-fertilizer application, 3. rotten straw application, and 4. a co-application of bio-fertilizer and rotten straw. Results showed that plant morphological characters of Baiyan2 were better than Caoyou1, and that soluble sugar and organic acid levels in the rhizosphere of Baiyan2 were significantly lower than Caoyou1. Compared to the control, oat root and plant development was significantly improved by the combined bio-fertilizer and rotten straw amendment. Bio-fertilizer application promoted malic and citric acid levels, contributing to a higher total organic acid level, and significantly increased the abundance of Rhizopus arrhizus and decreased the abundance of the fungal pathogens Alternaria, Cladosporium, Sarocladium and Heydenia of Ascomycota in both oat cultivars. All amendment treatments containing rotten straw, except the combined amendment in Baiyan2, significantly increased the relative abundance of Ascomycota (specifically Gibberella, Talaromyces, Fusarium, and Bipolaris) and decreased the relative abundance of R. arrhizus by reducing soluble sugar and organic acid levels. For the combined amendment in Baiyan2, there were no significant changes in Gibberella and Rhizopus between the control and amendment treatment. Our results suggest that co-application of bio-fertilizer and rotten straw, combined with a tolerant oat cultivar, is an effective method to increase crop productivity and enhance soil health in a saline-alkaline environment.
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Avena , Hongos , Micobioma , Raíces de Plantas , Rizosfera , Microbiología del Suelo , Suelo/química , Avena/anatomía & histología , Avena/crecimiento & desarrollo , Avena/microbiología , Fertilizantes , Hongos/clasificación , Hongos/crecimiento & desarrollo , Raíces de Plantas/anatomía & histología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiología , SalinidadRESUMEN
A novel series of indazole/indole derivatives were discovered as glucagon receptor (GCGR) antagonists through scaffold hopping based on two literature leads: MK-0893 and LY-2409021. Further structure-activity relationship (SAR) exploration and optimization led to the discovery of multiple potent GCGR antagonists with excellent pharmacokinetic properties in mice and rats, including low systemic clearance, long elimination half-life, and good oral bioavailability. These potent GCGR antagonists could be used for potential treatment of type II diabetes.
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Indazoles/química , Receptores de Glucagón/antagonistas & inhibidores , Humanos , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Estrogen-related receptor α (ERRα) is an orphan nuclear receptor that has been functionally implicated in the regulation of energy homeostasis. Herein is described the development of indazole-based N-alkylthiazolidenediones, which function in biochemical assays as selective inverse agonists against this receptor. Series optimization provided several potent analogues that inhibited the recruitment of a co-activator peptide fragment in vitro (IC50s < 50 nM) and reduced fasted circulating insulin and triglyceride levels in a sub-chronic pre-diabetic rat model when administered orally (10 mg/kg). A multi-parametric optimization strategy led to the identification of 50 as an advanced lead, which was more extensively evaluated in additional diabetic models. Chronic oral administration of 50 in two murine models of obesity and insulin resistance improved glucose control and reduced circulating triglycerides with efficacies similar to that of rosiglitazone. Importantly, these effects were attained without the concomitant weight gain that is typically observed with the latter agent. Thus, these studies provide additional support for the development of such molecules for the potential treatment of metabolic diseases.
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Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/farmacología , Indazoles/farmacología , Receptores de Estrógenos/antagonistas & inhibidores , Animales , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/química , Indazoles/administración & dosificación , Indazoles/química , Ligandos , Masculino , Ratones , Ratones Obesos , Estructura Molecular , Ratas , Ratas Sprague-Dawley , Ratas Zucker , Relación Estructura-Actividad , Receptor Relacionado con Estrógeno ERRalfaRESUMEN
Previously disclosed H4 receptor modulators, the triamino substituted pyridines and pyrimidines, contain a free primary amino (-NH2) group. In this Letter we demonstrate that an exocyclic amine (NH2) is not needed to maintain affinity, and also show a significant divergence in the SAR of the pendant diamine component. These des-NH2 azacycles also show a distinct functional spectrum, that appears to be influenced by the diamine component; in the case of the 1,3-amino pyrimidines, the preferred diamine is the amino pyrrolidine instead of the more common piperazines. Finally, we introduce 3,5-diamino pyridazines as novel histamine H4 antagonists.
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Piridinas/farmacología , Receptores Acoplados a Proteínas G/efectos de los fármacos , Receptores Histamínicos/efectos de los fármacos , Humanos , Piridinas/química , Receptores Histamínicos H4RESUMEN
BACKGROUND: Optical sensing is a potential tool to estimate plant N status, but soil water deficits may interefere with forming a clear relationship. A greenhouse study was conducted with oat plants treated with three water regimes and four N levels to determine whether optical sensing could be used to estimate leaf N and relative water content (RWC). RESULTS: Leaf N was strongly correlated with reflectance at 550 nm and at around 705 nm, and N treatments caused a red-edge peak shift to lower wavelength. The ratio of the first derivative reflectance at 741-696 nm (FDRE) was identified to be a good estimator of leaf N at jointing (R(2) = 0.90) and heading (R(2) = 0.86) stages across water treatments. Leaf N also had a stronger association with the red-edge position (REP) at both stages (R(2) = 0.83 and 0.78), or with the ratio R4 (R760/R550) at jointing (R(2) = 0.88), than with chlorophyll meter (SPAD) readings. Under water stress, the predictive accuracy of leaf N increased with these reflectance indices, but decreased using SPAD readings. CONCLUSION: The results indicate that specific reflectance indices of FDRE, REP and R4 may be used for a rapid and non-destructive estimation of oat plant N status over a range of water regimes.
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Avena/metabolismo , Luz , Nitrógeno/metabolismo , Hojas de la Planta/metabolismo , Tecnología de Sensores Remotos , Agua/metabolismo , Clorofila/metabolismo , SequíasRESUMEN
Bio-electrospraying (BES) is a technique for directly jetting living cells that has significant implications for tissue engineering and regenerative medicine. However, the effect of BES on human adipose-derived stem cells (hASCs) remains unknown. Here, we show that an hASC suspension was successfully electrosprayed via a continuous, stable and linearly directed electrospray at 10 kV and at 3 ml/h. Morphological observations and Trypan Blue and CCK-8 assays revealed that the cells remained viable and proliferated at a rate similar to that of the controls (0 kV). However, at 20 kV, BES became unstable and cell viability was reduced. Moreover, hASCs electrosprayed at 10 kV retained their multilineage potential, successfully differentiating into chondrogenic, osteogenic and neurogenic lineages. Thus, BES does not significantly affect cell morphology, viability or multipotency.
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Adipocitos/citología , Biotecnología/métodos , Ingeniería de Tejidos/métodos , Tejido Adiposo/citología , Biomarcadores/análisis , Biomarcadores/metabolismo , Diferenciación Celular , Proliferación Celular , Supervivencia Celular , Células Cultivadas , HumanosRESUMEN
BACKGROUND: Litopenaeus Vannamei, well known as pacific white shrimp, is the most popular shrimp in the world shrimp market. Identification and characterization of shrimp muscle regulatory genes are not only important for shrimp genetic improvement, but also facilitate comparative genomic tools for understanding of muscle development and regeneration. METHODOLOGY/PRINCIPAL FINDINGS: A novel mRNA encoding for a putative DNA-binding protein LvDBP23 was identified from Litopenaeus vannamei abdominal muscle cDNA library. The LvDBP23 cDNA contains 639 nucleotides of protein-coding sequence with deduced 212 amino acids of predicted molecular mass 23.32 kDa with glycine-rich domain at amino acid position 94-130. The mRNA sequence is successfully used for producing LvDBP23 recombinant protein in sf9 insect cell expression system. The expression of LvDBP23 mRNA is presented in abdominal muscle and swimming leg muscle, as well as other tissues including intestine, lymphoid and gill. The mRNA expression has the highest level in abdominal muscle in all tested tissues. LVDBP23 transcript during the molt cycle is highly expressed in the intermolt stage. In vitro nucleic acid-binding assays reveal that LvDBP23 protein can bind to both ssDNA and dsDNA, indicating its possible role of regulation of gene transcription. CONCLUSIONS/SIGNIFICANCE: We are the first to report a DNA-binding protein identified from the abdominal muscle tissue of marine shrimp L. Vannamei. Its high-level specific expression during the intermot stage suggests its role in the regulation of muscle buildup during the growth phase of shrimp molt cycle.
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Músculos Abdominales/metabolismo , Proteínas de Unión al ADN/metabolismo , ADN/metabolismo , Muda/fisiología , Proteínas Recombinantes/metabolismo , Secuencia de Aminoácidos , Animales , Organismos Acuáticos , Secuencia de Bases , Western Blotting , Clonación Molecular , ADN/genética , Proteínas de Unión al ADN/genética , Datos de Secuencia Molecular , Penaeidae/genética , Penaeidae/crecimiento & desarrollo , ARN Mensajero/genética , Proteínas Recombinantes/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de AminoácidoRESUMEN
Growth differentiation factor 11 (GDF11) is one of the significant genes that control skeletal formation. Knockout of GDF11 function causes abnormal patterning of the anterior/posterior axial skeleton. The mRNA of GDF11 is initially translated to a precursor protein that undergoes a proteolytic cleavage to generate the C-terminal peptide or mature GDF11, and the N-terminal peptide named GDF11 propeptide. The propeptide can antagonize GDF11 activity in vitro. To investigate the effects of GDF11 propeptide on GDF11 function in vivo, we generated transgenic mice that over-express the propeptide cDNA in skeletal tissue. The transgenic mice showed formation of extra ribs on the seventh cervical vertebra (C7) as a result of transformation of the C7 vertebra into a thoracic vertebra. The GDF11 propeptide transgene mRNA was detected in tail tissue in embryos and was highly expressed in tail and calvaria bones after birth. A high frequency of C7 rib formation was noticed in the transgenic mouse line with a high level of transgene expression. The anterior boundaries of Hoxa-4 and Hoxa-5 mRNA in situ expressions showed cranial shifts from their normal prevertebra locations in transgenic embryos. These results demonstrated significant effects of GDF11 propeptide transgene on vertebral formation, which are likely occurring through depressing GDF11 function and altered locations of Hoxa-4 and Hoxa-5 expression.
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Proteínas Morfogenéticas Óseas/genética , Huesos/embriología , Huesos/metabolismo , Vértebras Cervicales/anomalías , Factores de Diferenciación de Crecimiento/genética , Señales de Clasificación de Proteína/genética , Vértebras Torácicas/anomalías , Animales , Proteínas Morfogenéticas Óseas/antagonistas & inhibidores , Proteínas Morfogenéticas Óseas/metabolismo , Vértebras Cervicales/embriología , Femenino , Técnicas de Transferencia de Gen , Factores de Diferenciación de Crecimiento/antagonistas & inhibidores , Factores de Diferenciación de Crecimiento/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Transgénicos , Anomalías Musculoesqueléticas/embriología , Anomalías Musculoesqueléticas/genética , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Enfermedades de la Columna Vertebral/congénito , Enfermedades de la Columna Vertebral/embriología , Enfermedades de la Columna Vertebral/genética , Vértebras Torácicas/embriología , Regulación hacia Arriba/genéticaRESUMEN
A study concerning the mechanism of the LDA-mediated ortho and remote metalation of N,N-dialkyl-2-biphenyl carboxamides (e.g., 4a) is reported. On the basis of site-selective lithiation/electrophile quench experiments, including deuteration, the LDA metalation of 4 is proposed to involve initial amide-base complexation (CIPE) and equilibrium formation of 5, whose fast reaction with an in situ electrophile (TMSCl) to afford 6 prevents its equilibration with 7. In the absence of an electrophile, 5 undergoes equilibration via 4a with 7, whose fate is instantaneous cyclization to a stable tetrahedral carbinolamine oxide 8 which, only upon hydrolysis, affords fluorenone (3).
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Amidas/química , Compuestos de Bifenilo/química , Amidas/síntesis química , Compuestos de Bifenilo/síntesis química , Catálisis , Ciclización , Fluorenos/síntesis química , Fluorenos/química , Estructura MolecularRESUMEN
Myostatin is a dominant inhibitor of skeletal muscle development and growth. As transgenic over-expression of myostatin propeptide dramatically enhanced muscle mass, we hypothesized that administration of myostatin propeptide will increase muscle growth. In this study, the wild-type form of porcine myostatin propeptide and its mutated form at the cleavage site of metalloproteinases of BMP-1/TLD family were produced from insect cells. In vitro A204 cells reporter assays showed that both wild-type and the mutated propeptides depressed myostatin activity. The recombinant propeptides at four-fold myostatin concentration can effectively block myostatin function during co-incubation with A204 cells. In particular, the mutated propeptide appeared much more effective than wild-type propeptide over a long period during the in vitro co-incubation. Administration of the mutated propeptide to neonatal mice at the age of 11 and 18 days was tested and showed significant increase in growth performance by 11-15% from the age of 25 to 57 days (P < 0.05). The major skeletal muscles of mice that were injected with mutated propeptide were 13.5-24.8% heavier than the control group (P < 0.05) as a result of muscle fiber hypertrophy. In conclusion, administration of the mutated myostatin propeptide during the neonatal period is an effective way for promoting muscle growth.
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Músculo Esquelético , Mutación , Miostatina , Animales , Animales Recién Nacidos , Línea Celular , Femenino , Humanos , Insectos , Masculino , Ratones , Músculo Esquelético/anatomía & histología , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/crecimiento & desarrollo , Miostatina/genética , Miostatina/farmacología , Precursores de Proteínas/genética , Precursores de Proteínas/farmacología , PorcinosRESUMEN
BACKGROUND: Skeletal muscle growth and maintenance are essential for human health. One of the muscle regulatory genes, namely myostatin, a member of transforming growth factor-beta, plays a dominant role in the genetic control of muscle mass. Myostatin is synthesized as a precursor protein, which generates the N-terminal propeptide and the C-terminal mature myostatin peptide by a post-translational cleavage event. Previously, transgenic over-expression of myostatin propeptide in skeletal muscle results in significant muscle growth in early stages of development. The objectives of present study were to further characterize muscle growth in later stages of life and to identify genes and their expression patterns that are responsible for adult muscle build-up by myostatin propeptide. RESULTS: Immunohistochemical staining with an antibody to the N-terminus indicates a high level of myostatin propeptide present in the muscles of transgenic mice while there were no apparent differences in myostatin protein distribution in the muscle fibers between the transgenic and wild-type mice. Main individual muscles increased by 76-152% in the transgenic mice over their wild-type littermate mice at 12 months of age. A large number of nuclei were localized in the central and basal lamina of the myofibers in the transgenic mice as the number of nuclei per fiber and 100 microm(2) area was significantly higher in transgenic mice than wild-type mice. By systemic comparisons of global mRNA expression patterns between transgenic mice and wild-type littermates using microarray and qRT-PCR techniques, we have identified distinct gene expression patterns to support adult muscle build-up by myostatin propeptide, which are comprised of enhanced expressions of myogenic regulatory factors and extracelullar matrix components, and differentially down-regulated expressions of genes related to protein degradation and mitochondrial ATP synthesis. CONCLUSION: The results present a coordinated pattern of gene expressions for reduced energy utilization during muscle build-up in adult stage. Enhanced muscle buildup by myostatin propeptide is sustained by reduced ATP synthesis as a result of a decreased activity of protein degradation. Myostatin propeptide may have a therapeutic application to the treatment of clinical muscle wasting problems by depressing myostatin activity.
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Perfilación de la Expresión Génica , Desarrollo de Músculos/genética , Fibras Musculares Esqueléticas/metabolismo , Miostatina/genética , Precursores de Proteínas/genética , Animales , Regulación de la Expresión Génica , Masculino , Ratones , Ratones Transgénicos , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Complementario/genéticaRESUMEN
Small ubiquitin-like modifiers (SUMO) work in a similar way as ubiquitin to alter the biological properties of a target protein by conjugation. A shrimp SUMO cDNA named LvSUMO-1 was identified in Litopenaeus vannamei. LvSUMO-1 cDNA contains a coding sequence of 282 nucleotides with untranslated regions of 37 bp at 5'-end and 347 bp at 3'-end, respectively. The deduced 93 amino acids exhibit 83% identity with the Western Honeybee SUMO-1, and more than 65% homologies with human and mouse SUMO-1. LvSUMO-1 mRNA is expressed in most L. vannamei tissues with the highest level in hepatopancrease. The mRNA expression of LvSUMO-1 over development stages in L. Vammamei is distinguished by a low level in nauplius stage and relatively high level in postlarva stage with continuous expression until juvenile stage. The LvSUMO-1 protein and its conjugated proteins are detected in both cytoplasm and nucleus in several tissues. Interestingly, LvSUMO-1 mRNA levels are high in abdominal muscle during the premolt stage, wherein it has significant activities of protein degradation, suggesting its possible role in the regulation of shrimp muscle protein degradation.