Aging alters histone H3 lysine 4 methylation in mouse germinal vesicle stage oocytes

Decreasing oocyte competence with maternal aging is a major factor in mammalian infertility. One of the factors contributing to this infertility is changes to chromatin modifications, such as histone acetylation in old MII stage oocytes. Recent studies indicate that changes in histone acetylation at MII arise at the germinal vesicle (GV) stage. We hypothesised that histone methylation could also change in old GV oocytes. To test this hypothesis, we examined mono-, di- and trimethylation of histone H3 lysine 4 (H3K4 me1, me2 and me3, respectively) in young and older oocytes from 6–8- and 42–44-week-old mice, respectively. We found that H3K4 me2 and me3 decreased in older compared with young GV oocytes (100% vs 81% and 100% vs 87%, respectively; P < 0.05). H3K4 me2 later increased in older MII oocytes (21% vs 56%; P < 0.05). We also examined the expression of genes encoding the H3K4 demethylases lysine (K)-specific demethylase 1A (Kdm1a) and retinol binding protein 2 (Rbp2). Expression of Kdm1a increased at both the mRNA and protein levels in older GV oocytes, but decreased in older MII oocytes (P < 0.05), and was negatively correlated with H3K4 me2 levels. Conversely, expression of Rbp2 mRNA and protein decreased in older GV oocytes (P < 0.05), and this was not correlated with H3K4 me3 levels. Finally, we showed that inhibition of Kdm1a of older oocytes at the GV stage restored levels of H3K4 me2 at the MII stage to those seen in ‘young’ oocytes (41% vs 38%; P > 0.05). These results suggest that changes in expression of H3K4 me2 and Kdm1a in older GV oocytes may represent a molecular mechanism underlying human infertility caused by aging.

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Alopecia in Harlequin mutant mice is associated with reduced AIF protein levels and expression of retroviral elements

Mammalian Genome ◽

10.1007/s00335-020-09854-0 ◽

2020 ◽

Author(s):

Maik Hintze ◽

Sebastian Griesing ◽

Marion Michels ◽

Birgit Blanck ◽

Lena Wischhof ◽

...

Keyword(s):

Hair Growth ◽

Transcriptional Regulators ◽

Mutant Mice ◽

Wild Type ◽

Protein Levels ◽

Expression Of Genes ◽

Genes Encoding ◽

Keratinocyte Cell ◽

Apoptosis Inducing Factor ◽

Hair Cortex

AbstractWe investigated the contribution of apoptosis-inducing factor (AIF), a key regulator of mitochondrial biogenesis, in supporting hair growth. We report that pelage abnormalities developed during hair follicle (HF) morphogenesis in Harlequin (Hq) mutant mice. Fragility of the hair cortex was associated with decreased expression of genes encoding structural hair proteins, though key transcriptional regulators of HF development were expressed at normal levels. Notably, Aifm1 (R200 del) knockin males and Aifm1(R200 del)/Hq females showed minor hair defects, despite substantially reduced AIF levels. Furthermore, we cloned the integrated ecotropic provirus of the Aifm1Hq allele. We found that its overexpression in wild-type keratinocyte cell lines led to down-regulation of HF-specific Krt84 and Krtap3-3 genes without altering Aifm1 or epidermal Krt5 expression. Together, our findings imply that pelage paucity in Hq mutant mice is mechanistically linked to severe AIF deficiency and is associated with the expression of retroviral elements that might potentially influence the transcriptional regulation of structural hair proteins.

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The JmjC Domain Histone Demethylase Ndy1 Regulates Redox Homeostasis and Protects Cells from Oxidative Stress

Molecular and Cellular Biology ◽

10.1128/mcb.00688-08 ◽

2008 ◽

Vol 28 (24) ◽

pp. 7451-7464 ◽

Cited By ~ 31

Author(s):

Christos Polytarchou ◽

Raymond Pfau ◽

Maria Hatziapostolou ◽

Philip N. Tsichlis

Keyword(s):

Redox Regulation ◽

Replicative Senescence ◽

Redox Homeostasis ◽

Histone H3 ◽

Quinone Oxidoreductase ◽

Expression Of Genes ◽

Genes Encoding ◽

Jmjc Domain ◽

Peptidase Inhibitor ◽

Induced Apoptosis

ABSTRACT The histone H3 demethylase Ndy1/KDM2B protects cells from replicative senescence. Changes in the metabolism of reactive oxygen species (ROS) are important for establishing senescence, suggesting that Ndy1 may play a role in redox regulation. Here we show that Ndy1 protects from H2O2-induced apoptosis and G2/M arrest and inhibits ROS-mediated signaling and DNA damage, while knockdown of Ndy1 has the opposite effects. Consistent with these observations, whereas Ndy1 overexpression promotes H2O2 detoxification, Ndy1 knockdown inhibits it. Ndy1 promotes the expression of genes encoding the antioxidant enzymes aminoadipic semialdehyde synthase (Aass), NAD(P)H quinone oxidoreductase-1 (Nqo1), peroxiredoxin-4 (Prdx4), and serine peptidase inhibitor b1b (Serpinb1b) and represses the expression of interleukin-19. At least two of these genes (Nqo1 and Prdx4) are regulated directly by Ndy1, which binds to specific sites within their promoters and demethylates promoter-associated histone H3 dimethylated at K36 and histone H3 trimethylated at K4. Simultaneous knockdown of Aass, Nqo1, Prdx4, and Serpinb1b in Ndy1-expressing cells to levels equivalent to those detected in control cells was sufficient to suppress the Ndy1 redox phenotype.

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Regulation of ABCA1 and ABCG1 gene expression in the intraabdominal adipose tissue

Biomeditsinskaya Khimiya ◽

10.18097/pbmc20166203283 ◽

2016 ◽

Vol 62 (3) ◽

pp. 283-289 ◽

Cited By ~ 2

Author(s):

V.V. Miroshnikova ◽

A.A. Panteleeva ◽

E.A. Bazhenova ◽

E.P. Demina ◽

T.S. Usenko ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Cholesterol Metabolism ◽

Nuclear Factor ◽

Specific Expression ◽

Protein Levels ◽

Expression Of Genes ◽

Genes Encoding ◽

And Control

Tissue specific expression of genes encoding cholesterol transporters ABCA1 and ABCG1 as well as genes encoding the most important transcriptional regulators of adipogenesis – LXRa, LXRb, PPARg and RORa has been investigated in intraabdominal adipose tissue (IAT) samples.A direct correlation between the content of ABCA1 and ABCG1 proteins with RORa protein level (r=0.480, p<0.05; r=0.435, p<0.05, respectively) suggests the role of the transcription factor RORa in the regulation of IAT ABCA1 and ABCG1 protein levels. ABCA1 and ABCG1 gene expression positively correlated with obesity indicators such as body mass index (BMI) (r=0.522, p=0.004; r=0.594, p=0.001, respectively) and waist circumference (r=0.403, p=0.033; r=0.474, p=0.013, respectively). The development of obesity is associated with decreased IAT levels of RORa and LXRb mRNA (p=0.016 and p=0.002, respectively). These data suggest that the nuclear factor RORa can play a significant role in the regulation of cholesterol metabolism and control IAT expression of ABCA1 and ABCG1, while the level of IAT LXRb gene expression may be an important factor associated with the development of obesity.

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Vitamin D-Related Genetics as Predictive Biomarker of Clinical Remission in Adalimumab-Treated Patients Affected by Crohn’s Disease: A Pilot Study

Pharmaceuticals ◽

10.3390/ph14121230 ◽

2021 ◽

Vol 14 (12) ◽

pp. 1230

Author(s):

Jessica Cusato ◽

Lorenzo Bertani ◽

Miriam Antonucci ◽

Cristina Tomasello ◽

Gian Paolo Caviglia ◽

...

Keyword(s):

Vitamin D ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Clinical Remission ◽

Predictive Biomarker ◽

Nucleotide Polymorphisms ◽

Protein Levels ◽

Expression Of Genes ◽

Genes Encoding ◽

Bowel Diseases

Adalimumab (ADA) is a human anti-tumor necrosis factor (TNF-α) monoclonal antibody used in inflammatory bowel diseases, such as Crohn’s disease (CD). Vitamin-D (VD) is important for biological functions, such as the modulation of expression of genes encoding enzymes and transporters involved in drug metabolism and transport. ADA trough levels were associated with VD concentrations in patients with IBD, but no data are present in the literature concerning VD pathway-related gene single-nucleotide polymorphisms (SNPs) in affecting clinical outcomes. For this reason, the aim of this study was to evaluate the ability of VD-related genetics to predict clinical remission at 3 and 12 months in patients affected by CD treated with ADA. Patients affected by CD were included in this study. SNPs in CYP27B1, CYP24A1, GC, and VDR genes were analyzed through real-time PCR. A total of 63 patients were enrolled. Calprotectin, hemoglobin, and C-reactive protein levels were influenced by SNPs in VDR, CYP27B1, and GC genes. After 3 months of therapy, clinical remission was predicted by smoke, systemic steroids, and VDR BsmI, whereas at 12 months by GC 1296AA/AC and VD supplementation. This study reports the association between VD pathway-related genetics and ADA treatment. Further studies are needed to confirm these promising data.

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Immunomodulatory Effects of Voriconazole on Monocytes Challenged with Aspergillus fumigatus: Differential Role of Toll-Like Receptors

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01018-07 ◽

2008 ◽

Vol 52 (9) ◽

pp. 3301-3306 ◽

Cited By ~ 44

Author(s):

Maria Simitsopoulou ◽

Emmanuel Roilides ◽

Fotini Paliogianni ◽

Christodoulos Likartsis ◽

John Ioannidis ◽

...

Keyword(s):

Protein Expression ◽

Aspergillus Fumigatus ◽

Immunomodulatory Effect ◽

Human Monocytes ◽

Necrosis Factor Alpha ◽

Protein Levels ◽

Expression Of Genes ◽

Tnf Α ◽

Genes Encoding ◽

Mrna And Protein Expression

ABSTRACT Voriconazole (VRC) has activity against Aspergillus fumigatus, the most frequent cause of invasive aspergillosis in immunocompromised patients. The combination of VRC and A. fumigatus hyphae induced a more pronounced profile of expression of genes encoding inflammatory molecules in human monocytes than Aspergillus alone did. Herein, we provide further evidence of the potential mechanism underlying this immunomodulatory effect of VRC on human monocytes in response to A. fumigatus hyphae. A significant additive antifungal effect was shown when VRC was combined with monocytes against A. fumigatus hyphae. Both A. fumigatus hyphae and VRC induced pronounced profiles of mRNA and protein expression of Toll-like receptor 2 (TLR2) as well as tumor necrosis factor alpha (TNF-α) in THP-1 monocytic cells compared to untreated cells. The VRC-induced increase was greater than that induced by hyphae. The combination of VRC and hyphae increased mRNA and protein expression of TLR2 and TNF-α to even higher levels than did either VRC or hyphae alone. In contrast, TLR4 expression, both at the mRNA and protein levels, was not increased by either VRC or hyphae or their combination. In addition, significantly more NF-κB was translocated to the nuclei of THP-1 cells treated with VRC than untreated cells. While VRC induced more NF-κB than hyphae did, treatment with the combination of the two factors induced the greatest NF-κB expression. The pronounced profile of TLR2 signaling, TNF-α expression, and NF-κB activation in the presence of VRC suggests an immunomodulatory effect leading to a more efficient response to A. fumigatus.

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Cocaine Administration and Its Withdrawal Enhance the Expression of Genes Encoding Histone-Modifying Enzymes and Histone Acetylation in the Rat Prefrontal Cortex

Neurotoxicity Research ◽

10.1007/s12640-017-9728-7 ◽

2017 ◽

Vol 32 (1) ◽

pp. 141-150 ◽

Cited By ~ 13

Author(s):

Anna Sadakierska-Chudy ◽

Małgorzata Frankowska ◽

Joanna Jastrzębska ◽

Karolina Wydra ◽

Joanna Miszkiel ◽

...

Keyword(s):

Prefrontal Cortex ◽

Histone Acetylation ◽

Cocaine Administration ◽

Expression Of Genes ◽

Genes Encoding ◽

Histone Modifying Enzymes

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Are the levels of Cdk4 and Cx43 proteins of porcine oocytes associated with follicular size?

Animal Biology ◽

10.1163/157075511x566533 ◽

2011 ◽

Vol 61 (2) ◽

pp. 211-224 ◽

Cited By ~ 3

Author(s):

Klaus-Peter Brüssow ◽

Bartosz Kempisty ◽

Paweł Antosik ◽

Margarita Lianeri ◽

Magdalena Woźna ◽

...

Keyword(s):

Western Blot ◽

Zona Pellucida ◽

Blot Analysis ◽

Western Blot Analysis ◽

Protein Levels ◽

Expression Of Genes ◽

Genes Encoding ◽

Translocation Mechanism ◽

Follicular Size ◽

Follicle Size

AbstractGap junction connections are formed by proteins which play an important role in oocyte developmental competency but there is little information on the relationship between follicle size and the expression of genes encoding these proteins. The aim of this study was to investigate the potential association between follicle size and the levels of Cdk4 and Cx43 proteins using western blot analysis and confocal microscopic observations. Cumulus-oocyte complexes (COCs) were collected from puberal gilts (n = 20) of large (>5 mm), medium (3-5 mm), and small (<3 mm) follicles, and stained with BCB. BCB+ COCs, which had finished their growth phase, were cultured in TCM 199 for 44 h. Western blot analysis revealed an increased level of Cdk4 protein in oocytes isolated from large follicles as compared to medium (P < 0.05) and small (P < 0.01) ones. We did not detect differences in Cx43 protein levels in oocytes collected from any follicle class. Confocal microscopic observation revealed a specific membrane and zona pellucida localization of Cdk4 protein in oocytes isolated from large follicles, but an exclusively cytoplasmatic distribution of Cdk4 in oocytes from smaller follicle categories. The effect of follicular size on Cdk4 is indicated by the higher level of Cdk4 protein in oocytes isolated from large follicles and its variable distribution – perhaps resulting from a specific translocation mechanism – in the membrane, zona pellucida, and cytoplasm. IVM may also have a significant effect on Cdk4, as seen from the considerable difference in the expression and localization of Cdk4 protein in oocytes after IVM.

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Measurement of glyoxalase gene expression

Biochemical Society Transactions ◽

10.1042/bst20140026 ◽

2014 ◽

Vol 42 (2) ◽

pp. 495-499 ◽

Cited By ~ 5

Author(s):

Mingzhan Xue ◽

Naila Rabbani ◽

Paul J. Thornalley

Keyword(s):

Gene Expression ◽

Protein Quantification ◽

Glyoxalase System ◽

Protein Levels ◽

Glyoxalase Ii ◽

Reverse Transcription Pcr ◽

Expression Of Genes ◽

Genes Encoding ◽

Common Technique ◽

The Common

The glyoxalase system is an important component of the enzymatic defence against glycation, preventing particularly quantitatively and functionally important glycation of protein and DNA by methylglyoxal. Expression of genes encoding Glo1 (glyoxalase I) and Glo2 (glyoxalase II) may be induced or suppressed, and rates of proteolysis of Glo1 and Glo2 proteins may change in health and disease. Quantitative assessment of glyoxalase gene expression at the mRNA and protein levels has become a key part of glyoxalase system characterization. For mRNA, there is the common technique of real-time RT (reverse transcription)–PCR and direct quantification of mRNA copy number by the Nanostring™ method. For glyoxalase protein quantification, there is the commonly used Western blotting, and also immunoassay and, in proteome-wide studies, quantitative proteomics and proteome dynamics. We provide protocols for the common methods below and briefly review their application.

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