scholarly journals Impaired expression of genes coding for reactive oxygen species scavenging enzymes in testes of Mtfr1/Chppr-deficient mice

Reproduction ◽  
2007 ◽  
Vol 134 (3) ◽  
pp. 483-492 ◽  
Author(s):  
Massimiliano Monticone ◽  
Laura Tonachini ◽  
Sara Tavella ◽  
Paolo Degan ◽  
Roberta Biticchi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxidative Dna Damage ◽  
Mitochondrial Protein ◽  
Nuclear Gene ◽  
Cell Types ◽  
Expression Of Genes ◽  
Gene Coding ◽  
Scavenging Enzymes ◽  
Deficient Mice

Mtfr1/Chppris a nuclear gene coding for a mitochondrial protein capable of inducing fission of this organelle in a sequence-specific manner. Here we show that in mice,Mtfr1/Chppris ubiquitously expressed and displays the highest level of expression in pubertal and adult testes and in particular in spermatids and Leydig cells. To investigateMtfr1functionin vivo, we analyzed homozygous mice null for this gene obtained through a gene trap approach. We show that these mice fail to expressMtfr1and that in their testes several genes coding for enzymes involved in the defense against oxidative stress are downregulated. Among these, we studied in particular glutathione peroxidase 3 and show its expression in selected testis cell types. Furthermore, we demonstrate oxidative DNA damage specifically in testes ofMtfr1-deficient mice likely resulting from a reduced antioxidant activity. As a whole, these data suggest thatMtfr1protects the male gonads against oxidative stress.

scholarly journals Heme Oxygenase-1 Deficiency and Oxidative Stress: A Review of 9 Independent Human Cases and Animal Models

2021 ◽  
Vol 22 (4) ◽  
pp. 1514 ◽  
Author(s):  
Akihiro Yachie
Keyword(s):  
Oxidative Stress ◽  
Animal Models ◽  
Heme Oxygenase ◽  
Inflammatory Diseases ◽  
Cell Types ◽  
Pharmacological Intervention ◽  
Heme Oxygenase 1 ◽  
Inflammatory Reactions

Since Yachie et al. reported the first description of human heme oxygenase (HO)-1 deficiency more than 20 years ago, few additional human cases have been reported in the literature. A detailed analysis of the first human case of HO-1 deficiency revealed that HO-1 is involved in the protection of multiple tissues and organs from oxidative stress and excessive inflammatory reactions, through the release of multiple molecules with anti-oxidative stress and anti-inflammatory functions. HO-1 production is induced in vivo within selected cell types, including renal tubular epithelium, hepatic Kupffer cells, vascular endothelium, and monocytes/macrophages, suggesting that HO-1 plays critical roles in these cells. In vivo and in vitro studies have indicated that impaired HO-1 production results in progressive monocyte dysfunction, unregulated macrophage activation and endothelial cell dysfunction, leading to catastrophic systemic inflammatory response syndrome. Data from reported human cases of HO-1 deficiency and numerous studies using animal models suggest that HO-1 plays critical roles in various clinical settings involving excessive oxidative stress and inflammation. In this regard, therapy to induce HO-1 production by pharmacological intervention represents a promising novel strategy to control inflammatory diseases.

Abstract 424: Vascular Smooth Muscle Cell Expression of S100a12 in Vivo Induces Enhanced Oxidative Stress & Vascular Remodeling

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Marion Hofmann Bowman ◽  
Jeannine Wilk ◽  
Gene Kim ◽  
Yanmin Zhang ◽  
Jalees Rehman ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Smooth Muscle ◽  
Vascular Remodeling ◽  
Oxidative Dna Damage ◽  
Fold Increase ◽  
Brdu Incorporation ◽  
Elastic Fibers ◽  
Nuclear Staining

S100A12 is a small calcium binding protein that is a signal transduction ligand of the receptor for advance glycation endproducts (RAGE). S100A12, like RAGE, is expressed in the vessel wall of atherosclerotic vasculature, particularly in smooth muscle cells (SMC). While RAGE has been extensively implicated in inflammatory states such as atherosclerosis, the role of S100A12 is less clear. We tested the hypothesis that expression of human S100A12 directly exacerbates vascular inflammation. Several lines of Bl6/J transgenic mice (tg) expressing human S100A12 in SMC under control of the SM22a promoter were generated. Primary aortic SMC from tg and wild type (wt) littermates were isolated and analyzed for (i) proliferation using MTS/Formazan Assay and BrdU incorporation, (ii) oxidative stress using using flow cytometry with MitoSOX antibody, oxidative DNA damage using immunofluorescence microscopy with anti-8-oxo-dG antibody, and NF-kB activation measured by EMSA and (iii) cytokine expression measured by IL-6 ELISA. Furthermore, the aortas from tg and wt mice were examined. Results: Tg but not wt SMC expressed S100A12 protein. Tg SMC had a significant 1.9 to 2.7 fold increase in conversion of MTS into Formazan at 24–96 hours likely reflective of increased metabolic activity since BrdU incorporation into DNA was less in tg compared to wt SMC (4% vs 21% positive BrdU nuclei, p <0.05). Tg SMC showed significantly higher levels of mitochondrial generated ROS, nuclear staining for oxidative DNA damage which was not detected in the nuclei of wt SMC’s, and a 2.5 fold increase in NFkB activity. IL-6 production at baseline was higher in tg SMC’s (615 vs 213 pg/ml, p< 0.05) and increased dramatically after LPS treatment (10 ng/ml) in tg SMC’s (2130 vs 415 pg/ml). Histologic examination of the thoracic aorta at 10 weeks of age revealed increased collagen deposition in the aortic media with fragmentation and disarray of elastic fibers. In vivo ultrasound revealed a progressive dilation of the aortic arch from age 10 weeks to 16 weeks of age (1.27 to 1.60 mm, p<0.05) in tg but not in wt littermate mice (1.30 to 1.33 mm, p=0.1). These data reveal the novel finding that targeted expression of human S100A12 in SMC modulates oxidative stress, inflammation and vascular remodeling.

scholarly journals Differences in the products of mitochondrial protein synthesis in vivo in human and mouse cells and their potential use as markers for the mitochondrial genome in human–mouse cell hybrids

1974 ◽  
Vol 144 (1) ◽  
pp. 161-164 ◽  
Author(s):  
Alec Jeffreys ◽  
Ian Craig
Keyword(s):  
Protein Synthesis ◽  
Mitochondrial Protein ◽  
Detailed Comparison ◽  
Cell Types ◽  
Mouse Cell ◽  
Human Cells ◽  
Mitochondrial Protein Synthesis ◽  
Cell Hybrids ◽  
Different Cell Types

The proteins synthesized in the mitochondria of mouse and human cells grown in tissue culture were examined by electrophoresis in polyacrylamide gels. The proteins were labelled by incubating the cells in the presence of [35S]methionine and an inhibitor of cytoplasmic protein synthesis (emetine or cycloheximide). A detailed comparison between the labelled products of mouse and human mitochondrial protein synthesis was made possible by developing radioautograms after exposure to slab-electrophoresis gels. Patterns obtained for different cell types of the same species were extremely similar, whereas reproducible differences were observed on comparison of the profiles obtained for mouse and human cells. Four human–mouse somatic cell hybrids were examined, and in each one only components corresponding to mouse mitochondrially synthesized proteins were detected.

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5372-5372
Author(s):  
Alvaro A Elorza ◽  
Brigham B Hyde ◽  
Hanna Mikkola ◽  
Sheila Collins ◽  
Orian S Shirihai
Keyword(s):  
Cell Proliferation ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Mitochondrial Protein ◽  
Erythroid Progenitor ◽  
Heme Synthesis ◽  
Erythroid Progenitor Cells ◽  
Deficient Mice

Abstract UCP2, an inner membrane mitochondrial protein, has been implicated in bioenergetics and Reactive Oxygen Species (ROS) modulation. UCP2 has been previously hypothesized to function as a facilitator of heme synthesis and iron metabolism by reducing ROS production. While UCP2 has been found to be induced by GATA1 during erythroid differentiation its role in erythropoiesis in vivo or in vitro has not been reported thus far. Here we report on the study of UCP2 role in erythropoiesis and the hematologic phenotype of UCP2 deficient mouse. In vivo we found that UCP2 protein peaks at early stages of erythroid maturation when cells are not fully committed in heme synthesis and then becomes undetectable at the reticulocyte stage. Iron incorporation into heme was unaltered in erythroid cells from UCP2 deficient mice. While heme synthesis was not influenced by UCP2 deficiency, mice lacking UCP2 had a delayed recovery from chemically induced hemolytic anemia. Analysis of the erythroid lineage from bone marrow and fetal liver revealed that in the UCP2 deficient mice the R3 (CD71high/Ter119high) population was reduced by 24%. The count of BFU-E and CFU-E colonies, scored in an erythroid colony assay, was unaffected, indicating an equivalent number of early erythroid progenitor cells in both UCP2 deficient and control cells. Ex-vivo differentiation assay revealed that UCP2 deficient c-kit+ progenitor cells expansion was overall reduced by 14% with population analysis determining that the main effect is at the R3 stage. No increased rate of apoptosis was found indicating that expansion rather than cell death is being compromised. Reduced expansion of c-kit+ cells was accompanied by 30% reduction in the phosphorylated form of ERK, a ROS dependent cytosolic regulator of cell proliferation. Analysis of ROS in UCP2 null erythroid progenitors revealed altered distribution of ROS resulting in 14% decrease in cytosolic and 32% increase in mitochondrial ROS. Restoration of the cytosolic oxidative state of erythroid progenitor cells by the pro-oxidant Paraquat reversed the effect of UCP2 deficiency on cell proliferation in in vitro differentiation assays. Together, these results indicate that UCP2 is a regulator of erythropoiesis and suggests that inhibition of UCP2 function may contribute to the development of anemia.

Promoter Elements of vav Drive Transgene Expression In Vivo Throughout the Hematopoietic Compartment

Blood ◽  
1999 ◽  
Vol 94 (6) ◽  
pp. 1855-1863 ◽  
Author(s):  
Sarah Ogilvy ◽  
Donald Metcalf ◽  
Leonie Gibson ◽  
Mary L. Bath ◽  
Alan W. Harris ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Transgene Expression ◽  
Cell Types ◽  
Transcription Unit ◽  
Erythroid Cells ◽  
Promoter Elements ◽  
Specific Expression ◽  
Expression Of Genes ◽  
B And T Lymphocytes

To develop a method for targeting expression of genes to the full hematopoietic system, we have used transgenic mice to explore the transcriptional regulation of the vav gene, which is expressed throughout this compartment but rarely outside it. Previously, we showed that a cluster of elements surrounding its promoter could drive hematopoietic-specific expression of a bacterial lacZ reporter gene, but the expression was confined to lymphocytes and was sporadically silenced. Those limitations are ascribed here to the prokaryotic reporter gene. With a human CD4 (hCD4) cell surface reporter, the vav promoter elements drove expression efficiently and stably in virtually all nucleated cells of adult hematopoietic tissues but not notably in nonhematopoietic cell types. In multiple lines, hCD4 appeared on most, if not all, B and T lymphocytes, granulocytes, monocytes, megakaryocytes, eosinophils, and nucleated erythroid cells. Moreover, high levels appeared on both lineage-committed progenitors and the more primitive preprogenitors. In the fetus, expression was evident in erythroid cells of the definitive but not the primitive type. These results indicate that a prokaryotic sequence can inactivate a transcription unit and that the vavpromoter region constitutes a potent transgenic vector for the entire definitive hematopoietic compartment.

scholarly journals Dual Roles of Helicobacter pylori NapA in Inducing and Combating Oxidative Stress

2006 ◽  
Vol 74 (12) ◽  
pp. 6839-6846 ◽  
Author(s):  
Ge Wang ◽  
Yang Hong ◽  
Adriana Olczak ◽  
Susan E. Maier ◽  
Robert J. Maier
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Oxidative Dna Damage ◽  
Wild Type Mouse ◽  
Wild Type ◽  
Oxygen Intermediates ◽  
Physiological Analysis ◽  
H Pylori

ABSTRACT Neutrophil-activating protein (NapA) has been well documented to play roles in human neutrophil recruitment and in stimulating host cell production of reactive oxygen intermediates (ROI). A separate role for NapA in combating oxidative stress within H. pylori was implied by studies of various H. pylori mutant strains. Here, physiological analysis of a napA strain was the approach used to assess the iron-sequestering and stress resistance roles of NapA, its role in preventing oxidative DNA damage, and its importance to mouse colonization. The napA strain was more sensitive to oxidative stress reagents and to oxygen, and it contained fourfold more intracellular free iron and more damaged DNA than the parent strain. Pure, iron-loaded NapA bound to DNA, but native NapA did not, presumably linking iron levels sensed by NapA to DNA damage protection. Despite its in vitro phenotype of sensitivity to oxidative stress, the napA strain showed normal (like that of the wild type) mouse colonization efficiency in the conventional in vivo assay. By use of a modified mouse inoculation protocol whereby nonviable H. pylori is first inoculated into mice, followed by (live) bacterial strain administration, an in vivo role for NapA in colonization efficiency could be demonstrated. NapA is the critical component responsible for inducing host-mediated ROI production, thus inhibiting colonization by the napA strain. An animal colonization experiment with a mixed-strain infection protocol further demonstrated that the napA strain has significantly decreased ability to survive when competing with the wild type. H. pylori NapA has unique and separate roles in gastric pathogenesis.

scholarly journals Ex Vivo Enteroids Recapitulate In Vivo Citrulline Production in Mice

2018 ◽  
Vol 148 (9) ◽  
pp. 1415-1420 ◽  
Author(s):  
Xiaoying Wang ◽  
Yang Yuan ◽  
Inka C Didelija ◽  
Mahmoud A Mohammad ◽  
Juan C Marini
Keyword(s):  
Ex Vivo ◽  
Transcript Abundance ◽  
Mutant Mice ◽  
Ex Vivo Model ◽  
Endogenous Production ◽  
Gene Coding ◽  
Cycle Disorders ◽  
Deficient Mice

Abstract Background The endogenous production of arginine relies on the synthesis of citrulline by enteral ornithine transcarbamylase (OTC). Mutations in the gene coding for this enzyme are the most frequent cause of urea cycle disorders. There is a lack of correlation between in vivo metabolic function and DNA sequence, transcript abundance, or in vitro enzyme activity. Objective The goal of the present work was to test the hypothesis that enteroids, a novel ex vivo model, are able to recapitulate the in vivo citrulline production of wild-type (WT) and mutant mice. Methods Six-week-old male WT and OTC-deficient mice [sparse fur and abnormal skin (spf-ash) mutation] were studied. Urea and citrulline fluxes were determined in vivo, and OTC abundance was measured in liver and gut tissue. Intestinal crypts were isolated and cultured to develop enteroids. Ex vivo citrulline production and OTC abundance were determined in these enteroids. Results Liver OTC abundance was lower (mean ± SE: 0.16 ± 0.01 compared with 1.85 ± 0.18 arbitrary units; P < 0.001) in spf-ash mice than in WT mice, but there was no difference in urea production. In gut tissue, OTC was barely detectable in mutant mice; despite this, a lower but substantial citrulline production (67 ± 3 compared with 167 ± 8 µmol · kg−1 · h−1; P < 0.001) was shown in the mutant mice. Enteroids recapitulated the in vivo findings of a very low OTC content accompanied by a reduced citrulline production (1.07 ± 0.20 compared with 4.64 ± 0.44 nmol · µg DNA−1 · d−1; P < 0.001). Conclusions Enteroids recapitulate in vivo citrulline production and offer the opportunity to study the regulation of citrulline production in a highly manipulable system.

scholarly journals Reduced RNA turnover as a driver of cellular senescence

2021 ◽  
Vol 4 (3) ◽  
pp. e202000809
Author(s):  
Nowsheen Mullani ◽  
Yevheniia Porozhan ◽  
Adèle Mangelinck ◽  
Christophe Rachez ◽  
Mickael Costallat ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cellular Senescence ◽  
Cell Types ◽  
Rna Turnover ◽  
Viral Origin ◽  
Double Stranded Rna ◽  
Expression Of Genes ◽  
Inflammatory State ◽  
Rna Exosome ◽  
Rna Accumulation

Accumulation of senescent cells is an important contributor to chronic inflammation upon aging. The inflammatory phenotype of senescent cells was previously shown to be driven by cytoplasmic DNA. Here, we propose that cytoplasmic double-stranded RNA has a similar effect. We find that several cell types driven into senescence by different routes share an accumulation of long promoter RNAs and 3′ gene extensions rich in retrotransposon sequences. Accordingly, these cells display increased expression of genes involved in response to double stranded RNA of viral origin downstream of the interferon pathway. The RNA accumulation is associated with evidence of reduced RNA turnover, including in some cases, reduced expression of RNA exosome subunits. Reciprocally, depletion of RNA exosome subunit EXOSC3 accelerated expression of multiple senescence markers. A senescence-like RNA accumulation was also observed in cells exposed to oxidative stress, an important trigger of cellular senescence. Altogether, we propose that in a subset of senescent cells, repeat-containing transcripts stabilized by oxidative stress or reduced RNA exosome activity participate in driving and maintaining the permanent inflammatory state characterizing cellular senescence.

scholarly journals 8-Hydroxydeoxyguanosine (8-OHdG) levels in urinary samples of pesticide sprayers on exposure to organophosphorus pesticides

2015 ◽  
Vol 2 (3) ◽  
pp. 59 ◽  
Author(s):  
B. P. Mishra ◽  
Z. G. Badade ◽  
Bhupinder Kaur Anand ◽  
Jhansi Lakshmi Lingidi ◽  
Sapna Jaiswal
Keyword(s):  
Oxidative Stress ◽  
Oxidative Dna Damage ◽  
Organophosphorus Pesticides ◽  
Farm Workers ◽  
Control Group ◽  
Human Populations ◽  
Mean Values ◽  
Exposure Effect ◽  
Invasive Techniques

<p class="abstract"><strong><span lang="EN-US">Background: </span></strong><span lang="EN-US">Free radicals and other reactive species are constantly generated <em>in vivo</em> and cause oxidative damage to biomolecules. DNA is probably the most biologically significant target of oxidative attack. Among numerous types of oxidative DNA damage the formation of 8-hydroxyguanosine (8-OHdG) is a sensitive biomarker of oxidative stress, an adduct formed as a result of biochemical reaction between ROS and DNA. Chronic exposure to </span>Organophosphorus <span lang="EN-US">(OP) pesticides is implicated in many health conditions that result from the induction of oxidative stress, including cytogenetic damage. The main objective of the study was to evaluate the biochemical levels of 8-OHdG in spot urinary samples under the exposed OP pesticide sprayers and farm workers. </span></p><p class="abstract"><strong><span lang="EN-US">Methods:</span></strong><span lang="EN-US"> In this study, 51 male pesticide sprayers and 39 farm workers in the age group of 18-47 years having exposure  ranged from 3 to 15 years in duration were selected. The referents (n=31) were selected on the same criteria as well as they were never exposed to pesticides at any time. This study was conducted during the growing season (January, 2009 – September, 2010). The most commonly used OP pesticides like chlorpyriphos, Diazinon, Dimethioate, Monocrotofos etc., were used in this study. Urine samples from each participant were taken in sterile tubes and were stored at -20<sup>0</sup>C till analysed. The concentration of 8-OHdG in samples were analyzed using ELISA.<strong></strong></span></p><p class="abstract"><strong><span lang="EN-US">Results: </span></strong><span lang="EN-US">The urinary levels of 8-OHdG were found to be significantly higher in the farm workers and pesticide sprayers in contrast to the level observed in the control group (p&lt;0.05). When the data was analyzed in the exposed groups in relation to duration of exposure it was found that both the farm workers and sprayers who were exposed to OP pesticides for less than 5 years showed the maximum mean values of 8-OHdG in comparison to those exposed to for more than 10 years.</span></p><p class="abstract"><strong><span lang="EN-US">Conclusions:</span></strong><span lang="EN-US"> In view of this regular bio monitoring studies in target human populations are imperative necessary due to frequent changes in pesticide formulations and introduction of newer pesticides. Despite that several life style factors may influence the<strong> </strong>urinary concentrations of 8-OHdG but still this non-invasive bio-marker 8-OHdG is preferred over other invasive techniques to evaluate the environmental and occupational exposure effect of OP pesticides on the genotoxicity of the exposed workers.</span></p>

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