scholarly journals Transcriptomic analysis elucidates the molecular processes associated with hydrogen peroxide-induced diapause termination in Artemia-encysted embryos

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247160
Author(s):  
Bonien Chen ◽  
Tah-Wei Chu ◽  
Kuohsun Chiu ◽  
Ming-Chang Hong ◽  
Tsung-Meng Wu ◽  
...  

Treatment with hydrogen peroxide (H2O2) raises the hatching rate through the development and diapause termination of Artemia cysts. To comprehend the upstream genetic regulation of diapause termination activated by exterior H2O2 elements, an Illumina RNA-seq analysis was performed to recognize and assess comparative transcript amounts to explore the genetic regulation of H2O2 in starting the diapause termination of cysts in Artemia salina. We examined three groupings treated with no H2O2 (control), 180 μM H2O2 (low) and 1800 μM H2O2 (high). The results showed a total of 114,057 unigenes were identified, 41.22% of which were functionally annotated in at least one particular database. When compared to control group, 34 and 98 differentially expressed genes (DEGs) were upregulated in 180 μM and 1800 μM H2O2 treatments, respectively. On the other hand, 162 and 30 DEGs were downregulated in the 180 μM and 1800 μM H2O2 treatments, respectively. Cluster analysis of DEGs demonstrated significant patterns among these types of 3 groups. GO and KEGG enrichment analysis showed the DEGs involved in the regulation of blood coagulation (GO: 0030193; GO: 0050818), regulation of wound healing (GO:0061041), regulation of hemostasis (GO: 1900046), antigen processing and presentation (KO04612), the Hippo signaling pathway (KO04391), as well as the MAPK signaling pathway (KO04010). This research helped to define the diapause-related transcriptomes of Artemia cysts using RNA-seq technology, which might fill up a gap in the prevailing body of knowledge.

2018 ◽  
Vol 46 (1) ◽  
pp. 5 ◽  
Author(s):  
Nikolina Novakov ◽  
Vladislav Mandić ◽  
Brankica Kartalović ◽  
Bojana Vidović ◽  
Nenad Stojanac ◽  
...  

Background: Fungal infections can cause serious problems infecting fish eggs, especially unfertilized or dead eggs. In the past, this problem was solved by using very effective chemicals such as malachite green and formalin. But, due to its toxicity and carcinogenicity, malachite green was banned for use in fish intended for human consumption. Formalin also has been banned in most countries. Chemicals and drugs recommended for use to treat fungal infections are hydrogen peroxide, salt, potassium permanganate etc. The goal of the present study was to determine and compare the efficacy of antifungal effects of hydrogen peroxide and sodium chloride on brown trout eggs.Materials, Methods & Results: The experiment was conducted in the brown trout hatchery, Šipovo, Bosnia and Herzegovina. The experimental groups contained 500 and 1000 mg/L of hydrogen peroxide with 15 and 30 min of exposition; 1 and 2.5% of sodium chloride with 15 and 30 min of exposition and a negative control group (no chemical treatment). The treatment concentrations were calculated and prepared from hydrogen peroxide of 35% active ingredient, and sodium chloride (sterilized) of 100% active ingredient. Eggs for the study were spawned from 11 females and 4 males. The first treatment was performed on the fourth day, and each next treatment was performed at 3-day intervals. Six treatments were administered until the 19th day after the fertilization. The treatment of the eggs was provided until the eggs reached the eyed stage. The effectiveness of the chemical treatments was measured by a hatch rate. There was a significant difference between all treated groups and the negative control group (P < 0.05). Hydrogen peroxide with a concentration of 500 mg/L for 30 min was the most effective and demonstrated a higher hatching rate (75.7%). Sodium chloride treatments resulted in statistically significantly lower hatching rates than hydrogen peroxide treatments. The hatching rate in salt treatment with a concentration of 2.5% for 30 min was 27.3% lower than in hydrogen peroxide treatment with a concentration of 500 mg/L for 30 min.Discussion: Hydrogen peroxide is an effective antifungal, antibacterial and antiviral compound, and according to the Food and Drug Administration (FDA), hydrogen peroxide and salt are approved and classified as a low regulatory priority for the control of oomycetes on all species and life stages of fish. It is considered to be a very environmentally compatible chemical because it does not produce any toxic bioproducts when it decomposes. Hydrogen peroxide stood out as the best candidate substance for fungal control. The fact that the treatment with hydrogen peroxide at a concentration of 500 mg/L for 30 min was more effective than treatments at a concentration of 1000 mg/L can be explained by temperature dependency and treatment frequency of this chemical. Salt was not such an effective fungicide as hydrogen peroxide. When using salt, toxicity to the eggs should also be considered. Salt solutions may cause egg deaths at levels of 2.5% or higher. It is possible that high salinities have an inhibitory effect on the movement of fish embryo due to the high osmotic impact on the perivitelline layer. Thus, hydrogen peroxide has proven to be efficient, inexpensive, easy to use and environmentally safe in preventing fungal infections on brown trout eggs.


2020 ◽  
Author(s):  
Dawei Zhang ◽  
Wenjing Wu ◽  
Xin Huang ◽  
Ke Xu ◽  
Cheng Zheng ◽  
...  

Abstract Background: Chinese domestic pig breeds are reputed for pork quality, but their low ratio of lean-to-fat carcass weight decreases production efficiency. A better understanding of the genetic regulation network of SC fat tissue is necessary for the rational selection of Chinese domestic pig breeds. In the present study, SC adipocytes were isolated from Jiaxing Black pigs (a Chinese indigenous pig breed with redundant SC fat deposition) and Large White pigs (a lean-type pig breed with relatively low SC fat deposition) and the expression profiles of mRNAs and lncRNAs were compared by RNA-seq analysis to identify biomarkers correlated with the differences of SC fat deposition between the two breeds.Results: A total of 3,371 differentially expressed genes (DEGs) and 1,182 differentially expressed lncRNAs (DELs) were identified in SC adipocytes between Jiaxing Black (JX) and Large White (LW) pigs, which included 797 upregulated mRNAs, 2,574 downregulated mRNAs, 461 upregulated lncRNAs and 721 downregulated lncRNAs. Gene Ontology and KEGG pathway analyses revealed that the DEGs and DELs were mainly involved in the immune response, cell fate determination, PI3K-Akt signaling pathway and MAPK signaling pathway, which are known to be related to adipogenesis and lipid metabolism. The expression levels of DEGs and DELs according to the RNA-seq data were verified by quantitative PCR, which showed 81.8% consistency. The differences in MAPK pathway activity between JX and LW pigs was confirmed by western blot analysis, with <100-fold elevated p38 phosphorylation in JX pigs.Conclusions: This study offers a detailed characterization of mRNAs and lncRNAs in fat- and lean-type pig breeds. The activity of the MAPK signaling pathway was found to be associated with subcutaneous adipogenesis. These results greatly enhance our understanding of the molecular mechanisms regulating SC fat deposition in pigs.


2020 ◽  
Author(s):  
Tao Zhong ◽  
Cheng Wang ◽  
Jiangtao Hu ◽  
Xiaoyong Chen ◽  
Lili Niu ◽  
...  

Abstract Background: Rumen is an important digestive organ of ruminant. From fetal to adult stage, the morphology, structure and function of rumen have changed significantly. But the intrinsic genetic regulation is still limited. We previously reported a genome-wide expression profile of miRNAs in prenatal goat rumens. In the present study, we rejoined analyzed the transcriptomes of rumen miRNAs during prenatal (E60 and E135) and postnatal (D30 and D150) stages.Results: A total of 66 differentially expressed miRNAs (DEMs) were identified in the rumen tissues from D30 and D150 goats. Of these, 17 DEMs were consistently highly expressed in the rumens at the preweaning stages (E60, E135 and D30), while down-regulated at D150. Noteworthy, annotation analysis revealed that the target genes regulated by the DEMs were mainly enriched in MAPK signaling pathway, Jak-STAT signaling pathway and Ras signaling pathway. Interestingly, the expression of miR-148a-3p was significantly high in the embryonic stage and down-regulated at D150. The potential binding sites between miR-148a-3p and QKI were predicted by the TargetScan and verified by the dual luciferase report assay. The co-localization of miR-148a-3p and QKI was observed not in intestinal tracts but in rumen tissues by in situ hybridization. Moreover, the expression of miR-148a-3p in the epithelium was significantly higher than that in the other layers, suggesting that miR-148a-3p involve in the development of rumen epithelial cells by targeting QKI. Subsequently, miR-148a-3p inhibitor was found to induce the proliferation of GES-1 cells.Conclusions: Taken together, these results identified the DEMs involved in the development of rumen and provided an insight into the regulation mechanism of goat rumens during development.


Author(s):  
Xiang Sun ◽  
Zhijun Ren ◽  
Yixian Cun ◽  
Cai Zhao ◽  
Xianglin Huang ◽  
...  

Abstract Hippo-YAP signaling pathway functions in early lineage differentiation of pluripotent stem cells, but the detailed mechanisms remain elusive. We found that knockout (KO) of Mst1 and Mst2, two key components of the Hippo signaling in mouse embryonic stem cells (ESCs), resulted in a disruption of differentiation into mesendoderm lineage. To further uncover the underlying regulatory mechanisms, we performed a series of ChIP-seq experiments with antibodies against YAP, ESC master transcription factors and some characterized histone modification markers as well as RNA-seq assays using wild type and Mst KO samples at ES and day 4 embryoid body stage respectively. We demonstrate that YAP is preferentially co-localized with super-enhancer (SE) markers such as Nanog, Sox2, Oct4 and H3K27ac in ESCs. The hyper-activation of nuclear YAP in Mst KO ESCs facilitates the binding of Nanog, Sox2 and Oct4 as well as H3K27ac modification at the loci where YAP binds. Moreover, Mst depletion results in novel SE formation and enhanced liquid-liquid phase-separated Med1 condensates on lineage associated genes, leading to the upregulation of these genes and the distortion of ESC differentiation. Our study reveals a novel mechanism on how Hippo-YAP signaling pathway dictates ESC lineage differentiation.


Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4673-4673
Author(s):  
Shan Hua Zou ◽  
Xuejiao Zhang ◽  
Weiguang Wang ◽  
Yunfeng Cheng

Abstract Abstract 4673 Introduction Immune thrombocytopenic purpura (ITP) is an autoimmune disease characterized by low platelets and bleeding. The mechanism of ITP has historically been attributed to platelet autoantibody production and the resultant platelet destruction. More recent evidence suggests a multifactorial pathogenesis. Patients show impaired immune regulation manifested by increased proliferation of helper T lymphocytes and cytotoxic T lymphocytes from patients can lyse platelets in vitro. A complex picture of the immune processes involved in autoimmunity has emerged over the last decade with the identification and characterization of immunoregulatory elements (receptors, cytokines, and other signaling molecules) and cell trafficking patterns. Protein arrays are rapidly becoming established as a powerful means to detect proteins, monitor their expression levels, and investigate protein interactions and functions. Method During August 2009 to April 2010,17 newly diagnosed patients with ITP were enrolled. The mean platelet count at onset is 25*109/L. Seventeen health adults were as control group. We detected the changes of 507 kinds of cytokines, receptors and other signaling molecules in patients with ITP at onset, remission and relapse using the RayBio® Biotin Label-based Human Antibody Array I. The expression level differed by more than 2-fold from that observed in control group was as abnormal. We analyzed these proteins by cluster analysis. Results Compared with the healthy volunteers, the ITP patients at onset showed 35 elevated and 130 decreased proteins, the ITP patients at relapse showed 99 elevated and 189 decreased cytokines, the patients at remission showed 94 elevated and 144 decreased cytokines. This study shown that Th1 cytokines, JAK-STAT signaling pathway molecules, TGF-beta signaling pathway molecules, chemokine signaling pathway molecules, MAPK signaling pathway molecules and TLR signaling pathway molecules maybe closely related to the pathogenesis in ITP. Conclusion An understanding of the interplay of cytokines, receptors and signaling molecules in the breakdown of self-tolerance has brought to light unrecognized mechanisms of the autoimmune destruction of platelets in ITP and potential targets for future therapeutic advances. These identified proteins may constitute novel biomarkers of ITP and might hold great potential as surrogate markers for monitoring the patients with ITP. Disclosures: No relevant conflicts of interest to declare.


2021 ◽  
Author(s):  
Yuanyuan Tian ◽  
Jiao Zhao ◽  
Ju Huang ◽  
Haiying Zhang ◽  
Fushun Ni ◽  
...  

Abstract Background:Tumor endothelial cells (TECs) play an indispensable role in tumor growth and metastasis. Compared with normal endothelial cells (NECs), TECs exhibit unique phenotypic and functional heterogeneity in terms of metabolism, genetics, and transcriptomics. It is not only the key to coordinate tumor angiogenesis, but also an important factor of immune regulation in the tumor microenvironment. In recent years, the role of TECs in tumor metabolism and invasion has been continuously reported. However, the research on the mechanism behind the complex functions of TECs is still at the basic stage. We use Oxford Nanopore Technology (ONT) three-generation full-length transcriptome sequencing to detect all genetic structural changes in the transcriptome of mouse TECs 2H-11 and mouse NECs SVEC4-10.Results: In Tumor endothelial cells 2H-11,1847genes are up-regulated and 1202 genes are down-regulated. According to the Gene ontology (GO) enrichment analysis of differentially expressed genes (DEGs), we found that different functional trends related to metabolic processes, developmental processes, localization, immune system processes, and locomotion are the main reasons for the differences. DEGs are mainly enriched in signal pathways related to cancer, immunity and metabolism, involving Pathways in cancer,Antigen processing and presentation , Proteoglycans in cancer, Focal adhesion, MAPK signaling pathway ,Protein digestion and absorption,ECM-receptor interaction,PI3K-Akt signaling pathway and Glutathione metabolism. We also obtained the structural variation of transcripts such as alternative splicing, gene fusion, and alternative polyadenylation and accurately quantified the expression of the transcript. Some of our results have been confirmed in other documents. But other data have not been reported yet, which is the focus of our future exploration.Conclusion: We try to use transcriptomics and bioinformatics methods to characterize tumor endothelial cell-related genes and signaling pathways.It could help better understand the molecular mechanisms of tumor endothelial cells involved in tumorigenesis and development. DEGs in key pathways may be potential diagnostic markers or therapeutic targets of TECs. Our data also provide useful genetic resources for improving the genome and transcriptome annotations of TECs and NECs.


2021 ◽  
Author(s):  
XuJie Duan ◽  
Hui Yang ◽  
Liang Zhang ◽  
Huiping Li ◽  
Zhiwei Zhi Sun ◽  
...  

Abstract Background Ovine pulmonary adenomatosis (OPA) is a contagious lung epithelial tumor of sheep caused by jaagsiekte sheep retrovirus (JSRV), which causes severe economic losses for the sheep industry in the world. The specific oncogenic mechanism of JSRV is not yet clarified. Methods In this study, RNA was extracted from lung tissues of 3 naturally infected OPA cases and 3 healthy individuals for transcriptome sequencing (RNA-Seq). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to confirm the sequencing data. Immunohistochemistry (IHC) and western blot (WB) were performed to confirm the signaling pathway enriched by DEGs that was activated in naturally infected OPA cases. Cell viability, wound-healing, transwell and colony formation assays were performed to assess the cell malignant transformation of sheep trophoblast cells (STCs) transformed with JSRV- env lentivirus in vitro, and then WB was performed to confirm the signaling pathway that had been validated in the lung tissues. Results A total of 366 DEGs (154 up-regulated and 212 down-regulated) were identified by RNA-Seq of lung tissues of naturally infected OPA cases and healthy individuals. GO analysis showed that 366 DEGs were significantly enriched in 178 GO terms, including 114 biological processes, 19 cellular components and 45 molecular functions. KEGG analysis showed that the DEGs mainly enriched in cell proliferation, differentiation, apoptosis and migration, such as PI3K/Akt/mTOR, MAPK and Hippo signaling pathway, and Hippo signaling pathway has never been reported in naturally infected OPA cases. qRT-PCR results of 10 DEGs which were selected randomly were consistent with RNA-Seq results. The protein expression of Hippo signaling pathway were up-regulated in naturally infected OPA lung tissues. Cell viability, wound-healing, transwell and colony formation assays confirmed that JSRV- env lentivirus caused malignant transformation of STCs and JSRV Env increased the protein expression of Hippo signaling pathway. Conclusions This research first identified the changes in the transcriptome level of naturally infected OPA lung tissues. These data confirm that the Hippo signaling pathway is involved in the mechanism of OPA, clarify the interaction between Hippo signaling pathway and JSRV Env, provide further evidence for the tumorigenic mechanism of JSRV.


2018 ◽  
Vol 10 (6) ◽  
pp. 549-558 ◽  
Author(s):  
Lauren Rusnak ◽  
Cong Tang ◽  
Qi Qi ◽  
Xiulei Mo ◽  
Haian Fu

Abstract Apoptosis signal-regulating kinase 1 (ASK1) is an important mediator of the cell stress response pathways. Because of its central role in regulating cell death, the activity of ASK1 is tightly regulated by protein–protein interactions and post-translational modifications. Deregulation of ASK1 activity has been linked to human diseases, such as neurological disorders and cancer. Here we describe the identification and characterization of large tumor suppressor 2 (LATS2) as a novel binding partner for ASK1. LATS2 is a core kinase in the Hippo signaling pathway and is commonly downregulated in cancer. We found that LATS2 interacts with ASK1 and increases ASK1-mediated signaling to promote apoptosis and activate the JNK mitogen-activated protein kinase (MAPK). This change in MAPK signaling is dependent on the catalytic activity of ASK1 but does not require LATS2 kinase activity. This work identifies a novel role for LATS2 as a positive regulator of the ASK1–MKK–JNK signaling pathway and establishes a kinase-independent function of LATS2 that may be part of the intricate regulatory system for cellular response to diverse stress signals.


Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1951
Author(s):  
Tao Zhong ◽  
Cheng Wang ◽  
Jiangtao Hu ◽  
Xiaoyong Chen ◽  
Lili Niu ◽  
...  

The rumen is an important digestive organ of ruminants. From the fetal to adult stage, the morphology, structure and function of the rumen change significantly. However, the knowledge of the intrinsic genetic regulation of these changes is still limited. We previously reported a genome-wide expression profile of miRNAs in pre-natal goat rumens. In this study, we combined and analyzed the transcriptomes of rumen miRNAs during pre-natal (E60 and E135) and post-natal (D30 and D150) stages. A total of 66 differentially expressed miRNAs (DEMs) were identified in the rumen tissues from D30 and D150 goats. Of these, 17 DEMs were consistently highly expressed in the rumens at the pre-weaning stages (E60, E135 and D30), while down-regulated at D150. Noteworthy, annotation analysis revealed that the target genes regulated by the DEMs were mainly enriched in MAPK signaling pathway, Jak-STAT signaling pathway and Ras signaling pathway. Interestingly, the expression of miR-148a-3p was significantly high in the embryonic stage and down-regulated at D150. The potential binding sites of miR-148a-3p in the 3′-UTR of QKI were predicted by the TargetScan and verified by the dual luciferase report assay. The co-localization of miR-148a-3p and QKI through in situ hybridization was observed in the rumen tissues but not in the intestinal tracts. Moreover, the expression of miR-148a-3p in the epithelium was significantly higher than that in the other layers of the rumen, suggesting that miR-148a-3p is involved in the development of the rumen epithelial cells by targeting QKI. Subsequently, miR-148a-3p inhibitor was found to induce the proliferation of GES-1 cells. Taken together, our study identified DEMs involved in the development of the rumen and provides insights into the regulation mechanism of rumen development in goats.


2021 ◽  
Author(s):  
Yumeng Geng ◽  
Chunyu Lu ◽  
Guozhong Jin ◽  
Shuying Li ◽  
Yuqing Cui ◽  
...  

Abstract In order to explore the transcriptomics and proteomics targets and pathways of Salvia miltiorrhiza polysaccharides (SMPs) alleviating florfenicol (FFC)-induced liver injury in broilers,60 1-day-old broilers were randomly divided into 3 groups: control group ( GP1) was fed tap water, FFC model (GP2) was given tap water containing FFC 0.15 g/L, and SMPs treatment group (GP3) was given tap water containing FFC 0.15 g/L and SMPs 5 g/L.Starting from 1 day of age, the drug was administered continuously for 5 days. On the 6th day, blood was collected from the heart and the liver was taken. Then 3 chickens were randomly taken from each group, and their liver tissues were aseptically removed and placed in an enzyme-free tube. Using high-throughput mRNA sequencing and TMT-labeled quantitative proteomics technology, the transcriptome and proteome of the three groups of broiler liver were analyzed respectively. The results of the study showed that the liver tissue morphology of the chicks in the GP1 and GP3 groups was complete, and there were no obvious necrotic cells in the liver cells. The liver tissue cells in the GP2 group showed obvious damage, the intercellular space increased, and the liver cells showed extensive vacuolation and steatosis. Compared with the GP1 group, the daily gain of chicks in the GP2 group was significantly reduced (P < 0.0 5 or P < 0.01). Compared with the GP2 group, the GP3 group significantly increased the daily gain of chicks (P <0.0 5 or P <0.01). Compared with the GP1 group, the serum levels of ALT, AST, liver LPO, ROS and IL-6 in the GP2 group were significantly increased (P < 0.0 5 or P < 0.01), and the contents of T-AOC, GSH-PX, IL-4 and IL-10 in the liver were significantly decreased (P < 0.0 5 o r P < 0.01). After SMPs treatment, the serum levels of ALT, AST, liver LPO, ROS and IL-6 were significantly reduced (P < 0.0 5 or P < 0.01), and the contents of T-AOC, GSH-PX, IL-4 and IL-10 in the liver were significantly increased (P < 0.0 5 or P < 0.01). There were 380 mRNA and 178 protein differentially expressed between GP2 group and GP3 group. Part of DEGs was randomly selected for QPCR verification, and the expression results of randomly selected FABP1, SLC16A1, GPT2, AACS and other genes were verified by QPCR to be consistent with the sequencing results, which demonstrated the accuracy of transcriptation-associated proteomics sequencing. The results showed that SMPs could alleviate the oxidative stress and inflammatory damage caused by FFC in the liver of chicken and restore the normal function of the liver. SMPs may alleviate the liver damage caused by FFC by regulating the drug metabolism - cytopigment P450, PPAR signaling pathway, MAPK signaling pathway, glutathione metabolism and other pathways.


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