scholarly journals Inhibition of RNA synthesis during Scriptaid exposure enhances gene reprogramming in SCNT embryos

Reproduction ◽  
2019 ◽  
pp. 123-133
Author(s):  
Vitor Braga Rissi ◽  
Werner Giehl Glanzner ◽  
Mariana Priotto de Macedo ◽  
Lady Katerine Serrano Mujica ◽  
Karine Campagnolo ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Histone Deacetylases ◽  
Rna Synthesis ◽  
Embryo Quality ◽  
Blastocyst Stage ◽  
Histone Deacetylases Inhibitors ◽  
Expression Of Genes ◽  
Genome Activation ◽  
Relative Mrna Expression ◽  
Hdaci Treatment

Insufficient epigenetic reprogramming is incompatible with normal development of embryos produced by somatic cell nuclear transfer (SCNT), but treatment with histone deacetylases inhibitors (HDACi) enhances development of SCNT embryos. However, the mechanisms underpinning HDACi benefits in SCNT embryos remain largely uncharacterized. We hypothesized that, in addition to enhancing reprogramming, HDACi treatment may promote expression of genes not required for early development of SCNT embryos. To test this hypothesis, RNA synthesis was inhibited by treating bovine SCNT embryos with 5,6-dichlorobenzimidazole 1-β-D-ribofuranoside (DBR), which were concomitantly treated or not with Scriptaid (Scrip; an HDACi). Development to the blastocyst stage was significantly increased by treatment with Scrip alone (26.6%) or associated with DRB (28.6%) compared to Control (17.9%). The total number of nuclei was significantly improved only in embryos that were treated with both Scrip + DRB. Nuclear decondensation after SCNT was significantly increased by DRB treatment either alone or associated with Scrip. The relative mRNA expression, evaluated during the embryo genome activation (EGA) transition, revealed that some KDMs (KDM1A, KDM3A, KDM4C and KDM6A) and DNMT1 where prematurely expressed in Scrip-treated embryos. However, treatment with Scrip + DRB inhibited early mRNA expression of those genes, as well as several other KDMs (KDM4A, KDM4B, KDM5A, KDM5B, KDM5C and KDM7A) compared to embryos treated with Scrip alone. These findings revealed that HDACi improved development in SCNT embryos compared to Control, but altered the expression of genes involved in epigenetic regulation and did not improve embryo quality. Inhibition of RNA synthesis during HDACi treatment enhanced nuclear chromatin decondensation, modulated gene expression and improved SCNT embryo quality.

scholarly journals Expression of Adenoviral E1A in Transformed Cells as an Additional Factor of HDACi-Dependent FoxO Regulation

Cells ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 97
Author(s):  
Alisa Morshneva ◽  
Olga Gnedina ◽  
Tamara Marusova ◽  
Maria Igotti
Keyword(s):  
Histone Deacetylases ◽  
Transformed Cells ◽  
Histone Deacetylases Inhibitors ◽  
Forkhead Box ◽  
Foxo Protein ◽  
P300 Binding ◽  
Foxo Transcription Factors ◽  
The Relationship ◽  
Hdaci Treatment ◽  
Over Time

The adenoviral early region 1A (E1A) protein has proapoptotic and angiogenic activity, along with its chemosensitizing effect, making it the focus of increased interest in the context of cancer therapy. It was previously shown that E1A-induced chemosensitization to different drugs, including histone deacetylases inhibitors (HDACi), appears to be mediated by Forkhead box O (FoxO) transcription factors. In this study, we explore the relationship between E1A expression and the modulation of FoxO activity with HDACi sodium butyrate (NaBut). We show here that the basal FoxO level is elevated in E1A-expressing cells. Prolonged NaBut treatment leads to the inhibition of the FoxO expression and activity in E1A-expressing cells. However, in E1A-negative cells, NaBut promotes the transactivation ability of FoxO over time. A more detailed investigation revealed that the NaBut-induced decrease of FoxO activity in E1A-expressing cells is due to the NaBut-dependent decrease in E1A expression. Therefore, NaBut-induced inhibition of FoxO in E1A-positive cells can be overcome under unregulated overexpression of E1A. Remarkably, the CBP/p300-binding domain of E1Aad5 is responsible for stabilization of the FoxO protein. Collectively, these data show that the expression of E1A increases the FoxO stability but makes the FoxO level more sensitive to HDACi treatment.

scholarly journals Cytokine and Chemokine mRNA Expressions after Mycobacterium tuberculosis-Specific Antigen Stimulation in Whole Blood from Hemodialysis Patients with Latent Tuberculosis Infection

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 595
Author(s):  
Ji Young Park ◽  
Sung-Bae Park ◽  
Heechul Park ◽  
Jungho Kim ◽  
Ye Na Kim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mrna Expression ◽  
Whole Blood ◽  
Latent Tuberculosis Infection ◽  
Latent Tuberculosis ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Healthy Individuals ◽  
Relative Mrna Expression ◽  
Mrna Expression Levels

There have been few reports on the kinetics of hemodialyzed (HD) patients’ immune responses in latent tuberculosis infection (LTBI). Therefore, in the present study, messenger ribonucleic acid (mRNA) expression levels of nine immune markers were analyzed to discriminate between HD patients with LTBI and healthy individuals. Nine cytokines and chemokines were screened through relative mRNA expression levels in whole blood samples after stimulation with Mycobacterium tuberculosis (MTB)-specific antigens from HD patients with LTBI (HD/LTBI), HD patients without LTBI, and healthy individuals, and results were compared with the QuantiFERON-TB Gold In-Tube (QFT-GIT) test. We confirmed that the C-C motif chemokine 11 (CCL11) mRNA expression level of the HD/LTBI group was significantly higher than the other two groups. Especially, the CCL11 mRNA expression level of the >0.7 IU/mL group in the QFT-GIT test was significantly higher than the <0.2 IU/mL group in the QFT-GIT test and the 0.2–0.7 IU/mL group in the QFT-GIT test (p = 0.0043). The present study reveals that the relative mRNA expression of CCL11 was statistically different in LTBI based on the current cut-off value (i.e., ≥0.35 IU/mL) and in the >0.7 IU/mL group. These results suggest that CCL11 mRNA expression might be an alternative biomarker for LTBI diagnosis in HD patients.

scholarly journals Effects of Supplements Differing in Fatty Acid Profile to Late Gestational Beef Cows on Steer Progeny Finishing Phase Growth Performance, Carcass Characteristics, and mRNA Expression of Myogenic and Adipogenic Genes

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1904
Author(s):  
Taoqi Shao ◽  
Joshua C. McCann ◽  
Daniel W. Shike
Keyword(s):  
Fatty Acid ◽  
Growth Performance ◽  
Mrna Expression ◽  
Fatty Acid Profile ◽  
Carcass Characteristics ◽  
Beef Cows ◽  
Feed Ratio ◽  
Phase Growth ◽  
Pufa Supplementation ◽  
Relative Mrna Expression

The objective was to investigate the effects of feeding late gestational beef cows supplements differing in fatty acid profile on steer progeny finishing phase growth performance, carcass characteristics, and relative mRNA expression of myogenic and adipogenic genes. Seventy Angus-cross steers (initial body weight [BW] 273 ± 34 kg) born from dams supplemented with either 155 g DM/d EnerGII (CON, rich in palmitic and oleic acids) or 80 g DM/d Strata + 80 g DM/d Prequel (PUFA, rich in linoleic acid, eicosapentaenoic acid, and docosahexaenoic acid) for the last 77 ± 6 d prepartum were used. Longissimus muscle and subcutaneous adipose biopsies were collected to evaluate relative mRNA expression of genes related to myogenesis and adipogenesis. Steers were slaughtered at 423 ± 6 d of age. No treatment × time interaction or treatment effect (p ≥ 0.21) was detected for steer finishing phase BW, while steers from PUFA supplemented dams tended (p = 0.06) to have a greater gain to feed ratio (G:F). Neither carcass characteristics nor relative mRNA expression was different (p ≥ 0.11). In conclusion, late gestation PUFA supplementation tended to increase steer progeny finishing phase G:F, but had no effects on finishing phase BW, carcass characteristics, or relative mRNA expression during the finishing phase.

scholarly journals Analysis of Morphokinetic Parameters of Feline Embryos Using a Time-Lapse System

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 748
Author(s):  
Joanna Kochan ◽  
Agnieszka Nowak ◽  
Barbara Kij ◽  
Sylwia Prochowska ◽  
Wojciech Niżański
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Time Lapse ◽  
Blastocyst Stage ◽  
Cavity Formation ◽  
Non Invasive ◽  
Morphokinetic Parameters ◽  
Morphological Defects ◽  
Development In Vitro

The aim of this study was to analyze the morphokinetic parameters of feline embryos using a time lapse system. Oocytes matured in vitro were fertilized (IVF) and in vitro cultured in a time lapse-system (Primo Vision®, Gothenburg, Sweden). The first cell division of embryos occurred between 17 h post insemination (hpi) and 38 hpi, with the highest proportion of embryos (46%) cleaving between 21 and 24 hpi. The timing of the first cleavage significantly affected further embryo development, with the highest development occurring in embryos that cleaved at 21–22 hpi. Embryos that cleaved very early (17–18 hpi) developed poorly to the blastocyst stage (2%) and none of the embryos that cleaved later than 27 hpi were able to reach the blastocyst stage. Morphological defects were observed in 48% of the embryos. There were no statistically significant differences between the timing intervals of the first cleavage division and the frequency of morphological defects in embryos. Multiple (MUL) morphological defects were detected in more than half (56%) of the abnormal embryos. The most frequent single morphological defects were cytoplasmic fragmentation (FR) (8%) and blastomere asymmetry (AS) (6%). Direct cleavage (DC) from 1–3 or 3–5 blastomeres, reverse cleavage (RC) and vacuoles were rarely observed (2–3%). The timing of blastocyst cavity formation is a very good indicator of embryo quality. In our study, blastocyst cavity formation occurred between 127–167 hpi, with the highest frequency of hatching observed in blastocysts that cavitated between 142–150 hpi. Blastocysts in which cavitation began after 161 h did not hatch. In conclusion, the timing of the first and second cleavage divisions, the timing of blastocyst cavity formation and morphological anomalies can all be used as early and non-invasive indicators of cat embryo development in vitro.

8712 POSTER Validation of Differential mRNA Expression of Genes in Astrocytoma

2011 ◽  
Vol 47 ◽  
pp. S578
Author(s):  
C. Diez-Tascón ◽  
A. de la Hera ◽  
O. Rivero-Lezcano ◽  
L. Vilorio ◽  
E. Santin ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Expression Of Genes ◽  
Differential Mrna Expression

Abstract 191: NF-?B p65 Methylation by Protein Arginine Methyltransferase 5 (PRMT5) is Necessary for the Transcriptional Induction of CXCL10 by TNF-a

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Daniel P Harris
Keyword(s):  
Endothelial Cells ◽  
Mrna Expression ◽  
Chromatin Immunoprecipitation ◽  
Proximal Promoter ◽  
Protein Arginine Methyltransferase ◽  
Arginine Methyltransferase ◽  
Expression Of Genes ◽  
Tnf Α ◽  
Affect Expression ◽  
Transcriptional Induction

TNF-α initiates the expression of genes involved in the recruitment, adhesion, and transmigration of leukocytes to sites of inflammation. Here, we report that the protein arginine methyltransferase PRMT5 is required for the transcriptional induction of the pro-inflammatory chemokine CXCL10 (IP-10) in endothelial cells. Depletion of PRMT5 by siRNA results in significantly diminished TNF-α-induced CXCL10 mRNA expression, but does not affect expression of other chemokines, such as MCP-1 or IL-8. Chromatin immunoprecipitation experiments of the CXCL10 proximal promoter show the presence of symmetrical dimethylated arginine (sDMA)-containing proteins upon exposure to TNF-α. This methylation is completely lost when PRMT5 is removed from cells by siRNA. Using immunoprecipitation, we show that PRMT5 enhances CXCL10 expression by methylating the RelA (p65) subunit of NF-κB. In summary, we have identified that PRMT5 is a novel regulator of CXCL10 expression. Further, we have discovered that PRMT5 methylates NF-κB, a finding which may further knowledge of the post-translational code governing NF-κB regulation and target specificity.

Effects of lipotoxicity on a novel insulin-secreting human pancreatic β-cell line, 1.1B4

2013 ◽  
Vol 394 (7) ◽  
pp. 909-918 ◽  
Author(s):  
Srividya Vasu ◽  
Neville H. McClenaghan ◽  
Jane T. McCluskey ◽  
Peter R. Flatt
Keyword(s):  
Mrna Expression ◽  
Cell Line ◽  
Molecular Mechanisms ◽  
Endoplasmic Reticulum Stress Response ◽  
Future Research ◽  
The Novel ◽  
Pancreatic Β Cell ◽  
Β Cell ◽  
Expression Of Genes ◽  
Cellular Ca

Abstract The novel insulin-secreting human pancreatic β-cell line, 1.1B4, demonstrates stability in culture and many of the secretory functional attributes of human pancreatic β-cells. This study investigated the cellular responses of 1.1B4 cells to lipotoxicity. Chronic 18-h exposure of 1.1B4 cells to 0.5 mm palmitate resulted in decreased cell viability and insulin content. Secretory responses to classical insulinotropic agents and cellular Ca2+ handling were also impaired. Palmitate decreased glucokinase activity and mRNA expression of genes involved in secretory function but up-regulated mRNA expression of HSPA5, EIF2A, and EIF2AK3, implicating activation of the endoplasmic reticulum stress response. Palmitate also induced DNA damage and apoptosis of 1.1B4 cells. These responses were accompanied by increased gene expression of the antioxidant enzymes SOD1, SOD2, CAT and GPX1. This study details molecular mechanisms underlying lipotoxicity in 1.1B4 cells and indicates the potential value of the novel β-cell line for future research.

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