Melatonin Priming Alleviates Aging-Induced Germination Inhibition by Regulating β-oxidation, Protein Translation, and Antioxidant Metabolism in Oat (Avena sativa L.) Seeds

Although melatonin has been reported to play an important role in regulating metabolic events under adverse stresses, its underlying mechanisms on germination in aged seeds remain unclear. This study was conducted to investigate the effect of melatonin priming (MP) on embryos of aged oat seeds in relation to germination, ultrastructural changes, antioxidant responses, and protein profiles. Proteomic analysis revealed, in total, 402 differentially expressed proteins (DEPs) in normal, aged, and aged + MP embryos. The downregulated DEPs in aged embryos were enriched in sucrose metabolism, glycolysis, β-oxidation of lipid, and protein synthesis. MP (200 μM) turned four downregulated DEPs into upregulated DEPs, among which, especially 3-ketoacyl-CoA thiolase-like protein (KATLP) involved in the β-oxidation pathway played a key role in maintaining TCA cycle stability and providing more energy for protein translation. Furthermore, it was found that MP enhanced antioxidant capacity in the ascorbate-glutathione (AsA-GSH) system, declined reactive oxygen species (ROS), and improved cell ultrastructure. These results indicated that the impaired germination and seedling growth of aged seeds could be rescued to a certain level by melatonin, predominantly depending on β-oxidation, protein translation, and antioxidant protection of AsA-GSH. This work reveals new insights into melatonin-mediated mechanisms from protein profiles that occur in embryos of oat seeds processed by both aging and priming.

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Comparative Time-Course Physiological Responses and Proteomic Analysis of Melatonin Priming on Promoting Germination in Aged Oat (Avena sativa L.) Seeds

International Journal of Molecular Sciences ◽

10.3390/ijms22020811 ◽

2021 ◽

Vol 22 (2) ◽

pp. 811

Author(s):

Huifang Yan ◽

Peisheng Mao

Keyword(s):

Antioxidant Capacity ◽

Avena Sativa ◽

Proteomic Analysis ◽

Time Course ◽

Intact Cell ◽

Membrane Integrity ◽

Acid Synthesis ◽

Ultrastructural Changes ◽

Oat Seeds ◽

Aged Seeds

Melatonin priming is an effective strategy to improve the germination of aged oat (Avena sativa L.) seeds, but the mechanism involved in its time-course responses has remained largely unknown. In the present study, the phenotypic differences, ultrastructural changes, physiological characteristics, and proteomic profiles were examined in aged and melatonin-primed seed (with 10 μM melatonin treatment for 12, 24, and 36 h). Thus, 36 h priming (T36) had a better remediation effect on aged seeds, reflecting in the improved germinability and seedlings, relatively intact cell ultrastructures, and enhanced antioxidant capacity. Proteomic analysis revealed 201 differentially abundant proteins between aged and T36 seeds, of which 96 were up-accumulated. In melatonin-primed seeds, the restoration of membrane integrity by improved antioxidant capacity, which was affected by the stimulation of jasmonic acid synthesis via up-accumulation of 12-oxo-phytodienoic acid reductase, might be a candidate mechanism. Moreover, the relatively intact ultrastructures enabled amino acid metabolism and phenylpropanoid biosynthesis, which were closely associated with energy generation through intermediates of pyruvate, phosphoenolpyruvate, fumarate, and α-ketoglutarate, thus providing energy, active amino acids, and secondary metabolites necessary for germination improvement of aged seeds. These findings clarify the time-course related pathways associated with melatonin priming on promoting the germination of aged oat seeds.

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Metabolomics of aging in primary fibroblasts from small and large breed dogs

GeroScience ◽

10.1007/s11357-021-00388-0 ◽

2021 ◽

Author(s):

Paul S. Brookes ◽

Ana Gabriela Jimenez

Keyword(s):

Aging Process ◽

Tca Cycle ◽

Therapeutic Strategies ◽

Targeted Metabolomics ◽

The Tca Cycle ◽

Age Dependent ◽

Primary Fibroblasts ◽

Aging Rates ◽

Underlying Mechanisms ◽

Coenzyme A Biosynthesis

AbstractAmong several animal groups (eutherian mammals, birds, reptiles), lifespan positively correlates with body mass over several orders of magnitude. Contradicting this pattern are domesticated dogs, with small dog breeds exhibiting significantly longer lifespans than large dog breeds. The underlying mechanisms of differing aging rates across body masses are unclear, but it is generally agreed that metabolism is a significant regulator of the aging process. Herein, we performed a targeted metabolomics analysis on primary fibroblasts isolated from small and large breed young and old dogs. Regardless of size, older dogs exhibited lower glutathione and ATP, consistent with a role for oxidative stress and bioenergetic decline in aging. Furthermore, several size-specific metabolic patterns were observed with aging, including the following: (i) An apparent defect in the lower half of glycolysis in large old dogs at the level of pyruvate kinase. (ii) Increased glutamine anaplerosis into the TCA cycle in large old dogs. (iii) A potential defect in coenzyme A biosynthesis in large old dogs. (iv) Low nucleotide levels in small young dogs that corrected with age. (v) An age-dependent increase in carnitine in small dogs that was absent in large dogs. Overall, these data support the hypothesis that alterations in metabolism may underlie the different lifespans of small vs. large breed dogs, and further work in this area may afford potential therapeutic strategies to improve the lifespan of large dogs.

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Organic anion transporters 1 and 3 influence cellular energy metabolism in renal proximal tubule cells

Biological Chemistry ◽

10.1515/hsz-2018-0446 ◽

2019 ◽

Vol 400 (10) ◽

pp. 1347-1358 ◽

Cited By ~ 4

Author(s):

Jelle Vriend ◽

Charlotte A. Hoogstraten ◽

Kevin R. Venrooij ◽

Bartholomeus T. van den Berge ◽

Larissa P. Govers ◽

...

Keyword(s):

Proximal Tubule ◽

Organic Anion ◽

Depth Profiling ◽

Tca Cycle ◽

Oxidative Capacity ◽

Organic Anion Transporters ◽

Anion Transporters ◽

Oxidative Phenotype ◽

Underlying Mechanisms ◽

Lactate Levels

Abstract Organic anion transporters (OATs) 1 and 3 are, besides being uptake transporters, key in several cellular metabolic pathways. The underlying mechanisms are largely unknown. Hence, we used human conditionally immortalized proximal tubule epithelial cells (ciPTEC) overexpressing OAT1 or OAT3 to gain insight into these mechanisms. In ciPTEC-OAT1 and -OAT3, extracellular lactate levels were decreased (by 77% and 71%, respectively), while intracellular ATP levels remained unchanged, suggesting a shift towards an oxidative phenotype upon OAT1 or OAT3 overexpression. This was confirmed by increased respiration of ciPTEC-OAT1 and -OAT3 (1.4-fold), a decreased sensitivity to respiratory inhibition, and characterized by a higher demand on mitochondrial oxidative capacity. In-depth profiling of tricarboxylic acid (TCA) cycle metabolites revealed reduced levels of intermediates converging into α-ketoglutarate in ciPTEC-OAT1 and -OAT3, which via 2-hydroxyglutarate metabolism explains the increased respiration. These interactions with TCA cycle metabolites were in agreement with metabolomic network modeling studies published earlier. Further studies using OAT or oxidative phosphorylation (OXPHOS) inhibitors confirmed our idea that OATs are responsible for increased use and synthesis of α-ketoglutarate. In conclusion, our results indicate an increased α-ketoglutarate efflux by OAT1 and OAT3, resulting in a metabolic shift towards an oxidative phenotype.

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TrkB-Targeted Therapy for Mucoepidermoid Carcinoma

Biomedicines ◽

10.3390/biomedicines8120531 ◽

2020 ◽

Vol 8 (12) ◽

pp. 531

Author(s):

Vivian P. Wagner ◽

Manoela D. Martins ◽

Esra Amoura ◽

Virgilio G. Zanella ◽

Rafael Roesler ◽

...

Keyword(s):

Cell Migration ◽

Cell Lines ◽

Mucoepidermoid Carcinoma ◽

Combined Treatment ◽

Cell Ultrastructure ◽

Ultrastructural Changes ◽

Migration And Invasion ◽

Limiting Factor ◽

Transmission Electron

The brain-derived neurotrophic factor (BDNF)/tyrosine receptor kinase B (TrkB) pathway was previously associated with key oncogenic outcomes in a number of adenocarcinomas. The aim of our study was to determine the role of this pathway in mucoepidermoid carcinoma (MEC). Three MEC cell lines (UM-HMC-2, H253 and H292) were exposed to Cisplatin, the TrkB inhibitor, ANA-12 and a combination of these drugs. Ultrastructural changes were assessed through transmission electron microscopy; scratch and Transwell assays were used to assess migration and invasion; and a clonogenic assay and spheroid-forming assay allowed assessment of survival and percentage of cancer stem cells (CSC). Changes in cell ultrastructure demonstrated Cisplatin cytotoxicity, while the effects of ANA-12 were less pronounced. Both drugs, used individually and in combination, delayed MEC cell migration, invasion and survival. ANA-12 significantly reduced the number of CSC, but the Cisplatin effect was greater, almost eliminating this cell population in all MEC cell lines. Interestingly, the spheroid forming capacity recovered, following the combination therapy, as compared to Cisplatin alone. Our studies allowed us to conclude that the TrkB inhibition, efficiently impaired MEC cell migration, invasion and survival in vitro, however, the decrease in CSC number, following the combined treatment of ANA-12 and Cisplatin, was less than that seen with Cisplatin alone; this represents a limiting factor.

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Modifications in Tissue and Cell Ultrastructure as Elements of Immunity-Like Reaction in Chenopodium quinoa against Prune Dwarf Virus (PDV)

Cells ◽

10.3390/cells9010148 ◽

2020 ◽

Vol 9 (1) ◽

pp. 148 ◽

Cited By ~ 2

Author(s):

Edmund Kozieł ◽

Katarzyna Otulak-Kozieł ◽

Józef J. Bujarski

Keyword(s):

Vascular Tissue ◽

Chenopodium Quinoa ◽

Cell Ultrastructure ◽

Host Cells ◽

Dwarf Virus ◽

Ultrastructural Changes ◽

Viral Pathogen ◽

Prune Dwarf Virus ◽

Systemic Spread ◽

Specific Reactions

Prune dwarf virus (PDV) is a plant RNA viral pathogen in many orchard trees worldwide. Our knowledge about resistance genes or resistant reactions of plant hosts to PDV is scant. To fill in part of this gap, an aim of this study was to investigate reactions to PDV infection in a model host, Chenopodium quinoa. Our investigations concentrated on morphological and ultrastructural changes after inoculation with PDV strain 0599. It turned out that PDV infection can cause deformations in host cells but also induce changes in the organelles, such as chloroplasts in inoculated leaves. Moreover, we also demonstrated specific reactions/changes, which could be associated with both types of vascular tissue capable of effectively blocking the systemic spread of PDV to upper leaves. Furthermore, the relative amount of virus, P1 protein deposition, and movement protein (MP) gene expression consequently decreased in PDV-inoculated leaves.

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Sir2 is essential for intergenerational effects of parental diet on offspring metabolism in Drosophila

10.1101/641076 ◽

2019 ◽

Author(s):

Ryoya Hayashi ◽

Satomi Takeo ◽

Toshiro Aigaki

Keyword(s):

Fat Body ◽

Tca Cycle ◽

Primary Energy ◽

Standard Diet ◽

Wild Type ◽

Intergenerational Effects ◽

Transgenic Expression ◽

Fat Bodies ◽

Underlying Mechanisms ◽

High Level

AbstractRecent studies have revealed that parental diet can affect offspring metabolism and longevity in Drosophila. However, the underlying mechanisms are still unknown. Here we demonstrate that Sir2 encoding an NAD+-dependent histone deacetylase is required for the intergenerational effects of low nutrition diet (1:5 dilution of standard diet). We observed an increased amount of triacylglyceride (TAG) in the offspring when fathers were maintained on a low nutrition diet for 2 days. The offspring had increased levels of metabolites of glycolysis and TCA cycle, the primary energy producing pathways. We found that Sir2 mutant fathers showed no intergenerational effects. RNAi-mediated knockdown of Sir2 in the fat body was sufficient to mimic the Sir2 mutant phenotype, and the phenotype was rescued by transgenic expression of wild-type Sir2 in the fat body. Interestingly, even fathers had no experience of low nutrition diet, overexpression of Sir2 in their fat bodies induced a high level of TAG in the offspring. These findings indicated that Sir2 is essential in the fat body of fathers to induce intergenerational effects of low nutrition diet.

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HRI coordinates translation necessary for protein homeostasis and mitochondrial function in erythropoiesis

eLife ◽

10.7554/elife.46976 ◽

2019 ◽

Vol 8 ◽

Cited By ~ 17

Author(s):

Shuping Zhang ◽

Alejandra Macias-Garcia ◽

Jacob C Ulirsch ◽

Jason Velazquez ◽

Vincent L Butty ◽

...

Keyword(s):

Iron Deficiency ◽

Mitochondrial Function ◽

Ribosomal Proteins ◽

Target Genes ◽

Mitochondrial Protein ◽

Erythroid Differentiation ◽

Protein Translation ◽

Ribosome Profiling ◽

Underlying Mechanisms

Iron and heme play central roles in the production of red blood cells, but the underlying mechanisms remain incompletely understood. Heme-regulated eIF2α kinase (HRI) controls translation by phosphorylating eIF2α. Here, we investigate the global impact of iron, heme, and HRI on protein translation in vivo in murine primary erythroblasts using ribosome profiling. We validate the known role of HRI-mediated translational stimulation of integratedstressresponse mRNAs during iron deficiency in vivo. Moreover, we find that the translation of mRNAs encoding cytosolic and mitochondrial ribosomal proteins is substantially repressed by HRI during iron deficiency, causing a decrease in cytosolic and mitochondrial protein synthesis. The absence of HRI during iron deficiency elicits a prominent cytoplasmic unfolded protein response and impairs mitochondrial respiration. Importantly, ATF4 target genes are activated during iron deficiency to maintain mitochondrial function and to enable erythroid differentiation. We further identify GRB10 as a previously unappreciated regulator of terminal erythropoiesis.

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Activation of the TCA Cycle to Provide Immune Protection in Zebrafish Immunized by High Magnesium-Prepared Vibrio alginolyticus Vaccine

Frontiers in Immunology ◽

10.3389/fimmu.2021.739591 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jun Yang ◽

Xiao-li Yang ◽

Yu-bin Su ◽

Xuan-xian Peng ◽

Hui Li

Keyword(s):

Tca Cycle ◽

Dependent Manner ◽

High Concentration ◽

Immune Genes ◽

Bacterial Diseases ◽

The Tca Cycle ◽

Aquaculture Industry ◽

Innate Immune Genes ◽

Underlying Mechanisms ◽

Dose Dependent

Vaccines are safe and efficient in controlling bacterial diseases in the aquaculture industry and are in line with green farming. The present study develops a previously unreported approach to prepare a live-attenuated V. alginolyticus vaccine by culturing bacteria in a high concentration of magnesium to attenuate bacterial virulence. Furthermore, metabolomes of zebrafish immunized with the live-attenuated vaccines were compared with those of survival and dying zebrafish infected by V. alginolyticus. The enhanced TCA cycle and increased fumarate were identified as the most key metabolic pathways and the crucial biomarker of vaccine-mediated and survival fish, respectively. Exogenous fumarate promoted expression of il1β, il8, il21, nf-κb, and lysozyme in a dose-dependent manner. Among the five innate immune genes, the elevated il1β, il8, and lysozyme are overlapped in the vaccine-immunized zebrafish and the survival from the infection. These findings highlight a way in development of vaccines and exploration of the underlying mechanisms.

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Dynamic Microbial Shifts and Signatures of Long-Term Remission in Allergic Rhinitis After an Herbal Formula Treatment

Frontiers in Immunology ◽

10.3389/fimmu.2021.774966 ◽

2021 ◽

Vol 12 ◽

Author(s):

Libing Zhu ◽

Yuning Wu ◽

Chenglong Lin ◽

Lin Tang ◽

Bin Yu ◽

...

Keyword(s):

Allergic Rhinitis ◽

Tca Cycle ◽

Diversity Indices ◽

Sustained Remission ◽

Herbal Formula ◽

Long Term Effect ◽

Significant Difference ◽

Ppar Signaling ◽

Underlying Mechanisms

A mixed Chinese herbal formula, Xiao-Qing-Long-Decoction (XQLD), may contribute to sustained remission in allergic rhinitis (AR), but it is unknown which factors determine such long-term effect. Here, we aimed to identify bacterial signatures associated with sustained remission. To this end, samples from AR patients at four different times were analyzed to compare the dynamic bacterial community and structure shifts. Diversity indices Chao1 showed significant difference across different time (p<0.05), and the Kruskal-Wallis test identified that Dialister (OTU_31), Roseburia (OTU_36), Bacteroides (OTU_22), Bacteroides (OTU_2040), and Prevotella_9 (OTU_5) were the significant differential bacterial taxa (p<0.05). These distinctive genera were significantly associated with the change of AR clinical indices and the predicted functional pathways such as PPAR signaling pathway, peroxisome, and citrate cycle (TCA cycle) (p<0.05), indicating that they may be important bacterial signatures involving in the sustained remission in AR (p<0.05). Besides, lower Firmicutes/Bacteroidetes (F/B) ratio at 6 months follow-up may also contribute to the long-term remission of AR. No seriously adverse events and safety concerns were observed in this study. In conclusion, XQLD is a meaningful, long-term efficient and safe medication for AR treatment. The underlying mechanisms of sustained remission in AR after XQLD treatment may be associated with the dynamic alteration of featured gut bacteria taxa.

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IRS1 deficiency protects β-cells against ER stress-induced apoptosis by modulating sXBP-1 stability and protein translation

Scientific Reports ◽

10.1038/srep28177 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 10

Author(s):

Tomozumi Takatani ◽

Jun Shirakawa ◽

Michael W. Roe ◽

Colin A. Leech ◽

Bernhard F. Maier ◽

...

Keyword(s):

Er Stress ◽

Protein Translation ◽

Adaptor Proteins ◽

Nuclear Accumulation ◽

Cell Protection ◽

Β Cells ◽

Β Cell ◽

Protein Instability ◽

Underlying Mechanisms ◽

Induced Apoptosis

Abstract Endoplasmic reticulum (ER) stress is among several pathological features that underlie β-cell failure in the development of type 1 and type 2 diabetes. Adaptor proteins in the insulin/insulin-like-growth factor-1 signaling pathways, such as insulin receptor substrate-1 (IRS1) and IRS2, differentially impact β-cell survival but the underlying mechanisms remain unclear. Here we report that β-cells deficient in IRS1 (IRS1KO) are resistant, while IRS2 deficiency (IRS2KO) makes them susceptible to ER stress-mediated apoptosis. IRS1KOs exhibited low nuclear accumulation of spliced XBP-1 due to its poor stability, in contrast to elevated accumulation in IRS2KO. The reduced nuclear accumulation in IRS1KO was due to protein instability of Xbp1 secondary to proteasomal degradation. IRS1KO also demonstrated an attenuation in their general translation status in response to ER stress revealed by polyribosomal profiling. Phosphorylation of eEF2 was dramatically increased in IRS1KO enabling the β-cells to adapt to ER stress by blocking translation. Furthermore, significantly high ER calcium (Ca2+) was detected in IRS1KO β-cells even upon induction of ER stress. These observations suggest that IRS1 could be a therapeutic target for β-cell protection against ER stress-mediated cell death by modulating XBP-1 stability, protein synthesis, and Ca2+ storage in the ER.

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