173 Size Matters: a Follicle-of-Origin Effect on the Preovulatory Epigenetic Constitution of Germinal Vesicle-Stage Porcine Oocytes

2018 ◽  
Vol 30 (1) ◽  
pp. 226
Author(s):  
S. Heywood ◽  
R. L. Matheson ◽  
J. Thomas ◽  
L. Moley ◽  
S. C. Isom
Keyword(s):  
Multiple Testing ◽  
Germinal Vesicle ◽  
Nuclear Staining ◽  
Multiple Testing Correction ◽  
Histone Marks ◽  
Histone 3 ◽  
Germinal Vesicles ◽  
Preovulatory Follicles ◽  
Chromatin Configuration ◽  
Origin Effect

The goal of this study was to evaluate global levels of a variety of histone modifications at different lysine (K) residues on Histone 3 (H3) within the chromatin of porcine germinal vesicle (GV)-stage oocytes that were aspirated from follicles of different sizes. We hypothesised that we would see evidence of a transition from open, transcriptionally active chromatin (in oocytes from smaller, growing follicles) to more closed, transcriptionally silent chromatin associated with fully grown oocytes (aspirated from large, preovulatory follicles). Cumulus-enclosed oocytes were aspirated from small (<3 mm) or large (>7 mm) follicles from abattoir-derived pig ovaries. Oocytes were denuded immediately after aspiration and then immunoprobed with antibodies specific for trimethylated (me3) H3K4, H3K9me3, and H3K27me3. Background-corrected nuclear fluorescence levels for each histone mark were collected from multiple oocytes from each of at least three experimental replicates (aspiration days). Data were subjected to one-way ANOVA with a Bonferroni multiple testing correction to determine whether there were differences in fluorescence intensities in the nuclei (germinal vesicles) of oocytes from small v. large follicles. Oocytes from large follicles displayed more intense nuclear staining for all 3 histone marks: average nuclear H3K4me3 intensity was 31.4% higher (P = 0.0004), H3K9me3 was 70.3% higher (P = 0.0218), and H3K27me3 was 32.0% higher (P = 0.0231) in oocytes from large follicles. An ancillary analysis of the data revealed no effect (P > 0.1) of pubertal status (i.e. whether small and large follicles were aspirated from pre- v. post-pubertal ovaries) on the intensity of nuclear fluorescence for any of the marks evaluated. In continuation, 3 oocytes from both follicle types were collected on each of 6 aspiration days (i.e. 18 individual oocytes from each follicle type), and the mRNA from these were used for an RT-qPCR experiment to detect the relative abundance of transcripts from 21 different genes coding for histone methyltransferase or demethylase enzymes in oocytes from large v. small follicles. Of the 21 genes tested, 5 genes (KDM4C, KDM4D, KDM5B, KDM5C, and SETD7) were not detectable in our individual oocyte samples, but transcripts from 6 of the 16 remaining genes (KDM6A, KMT2B, MLL3, SETD1B, SETDB1, and SUV39H2) were shown to be significantly more abundant in oocytes from large follicles (at least 2-fold greater abundance and P < 0.05). Although our expectation-that histone marks (and related transcripts) would consistently reflect a globally “repressive” chromatin configuration in oocytes from large follicles and a more “open” configuration in oocytes from small follicles-turned out to be untrue, the evidence suggests that the epigenetic constitution of oocytes from small follicles may indeed vary from that of oocytes from large, preovulatory follicles, and this phenomenon warrants further investigation.

177 Collection of mouse immature oocytes using cilostazol, a phosphodiesterase 3A inhibitor treatment

2020 ◽  
Vol 32 (2) ◽  
pp. 216
Author(s):  
A. Taiyeb ◽  
R. A. Jassim ◽  
A. Alazzam ◽  
M. Kjelland ◽  
C. K. Mawlood ◽  
...  
Keyword(s):  
Conventional Method ◽  
Oral Treatment ◽  
Germinal Vesicle ◽  
Cortical Granules ◽  
Uniform Size ◽  
Immature Oocytes ◽  
Diameter Range ◽  
Preovulatory Follicles ◽  
Chromatin Configuration ◽  
N 93

Administration of cilostazol in mice and swine has resulted in the ovulation of immature oocytes at the germinal vesicle (GV) or metaphase I (MI) stages. The present study aimed to define oocyte synchronised maturation, yield, health, and ease of collection from mice treated with cilostazol. The conventional method included mice primed with pregnant mare serum gonadotrophin (PMSG) and GV oocytes isolated from preovulatory follicles 48h post-PMSG. Recovery of MI oocytes included the invitro maturation of the isolated GV oocytes into the MI stage for 6h or the superovulation of mice with PMSG and human chorionic gonadotrophin (hCG), 48h apart, and the isolation of MI oocytes from preovulatory follicles 6h post-hCG. The cilostazol method included the superovulation of mice, as described above, and oral treatment with 7.5mg of cilostazol once (at the same time as hCG) or twice (at the same time as hCG plus 6h post-hCG) to result in the ovulation of MI or GV oocytes, respectively. The cilostazol method resulted in immature oocytes that are uniform in size. For instance, the cilostazol method resulted in 98.0% (n=110) of GV oocytes with a diameter range of 60-90μm compared with only 49.5% (n=118) of GV oocytes resulting from the conventional method (P&lt;0.0001). Similarly, 95.0% (n=93) of MI oocytes obtained from the cilostazol method were synchronised within the diameter range of 50.1-70μm compared with 60.0% (n=89) of MI oocytes obtained from the conventional method (P&lt;0.0001). Cilostazol also resulted in immature oocytes with synchronised nuclear and cytoplasmic maturation. In this regard, the cilostazol method resulted in GV oocytes having higher levels of co-occurrence of peripheral cortical granules and surrounded nucleolus chromatin configuration compared with the conventional method (80.6% (n=124) vs. 36.6% (n=131), respectively; P&lt;0.0001). Similarly, the co-occurrence of normally organised spindles and chromosomes and peripheral cortical granules with free domains was observed more frequently in MI oocytes obtained from the cilostazol method than in those obtained from the conventional method (82.8% (n=151) vs. 65.0% (n=100), respectively; P=0.001). The cilostazol method was more time and labour efficient (0.8±0.2 vs. 3.2±0.2 min; P&lt;0.001) and resulted in higher oocyte yields (22.7±1.8 vs. 7.9±0.7 oocytes; P&lt;0.0001) and normal morphology (94±1.5% vs. 80.1±3.3%; P=0.02) than did the conventional method (P&lt;0.001). Finally, compared with the conventional method, the cilostazol method improved the blastocyst production rate of GV and MI oocytes from 39.6% (n=188) to 65.0% (n=169) and from 44.2% to 75.3%, respectively (P&lt;0.001). The presented method provides not only oocytes with uniform size and synchronised developmental maturation but also a technique of oocyte collection that is efficient and resourceful. It is possible that not all of the immature oocytes resulting from the conventional method are from preovulatory follicles, and they do not necessarily represent the cohort of oocytes that would develop adequately and consequently ovulate as opposed to the presented method.

scholarly journals Follicular development and preovulatory oocyte maturation in Hungarian Mangalica and Landrace gilts

2001 ◽  
Vol 44 (4) ◽  
pp. 413-420 ◽  
Author(s):  
I. Egerszegi ◽  
H. Torner ◽  
J. Rátky ◽  
K.-P. Brüssow
Keyword(s):  
Oocyte Maturation ◽  
Critical Level ◽  
Follicular Development ◽  
Germinal Vesicle ◽  
Follicular Growth ◽  
Physiological Basis ◽  
Meiotic Configuration ◽  
Preovulatory Follicles ◽  
Chromatin Configuration ◽  
Follicular Fluids

Abstract. Preservation of native pig breeds of different values has got increasing public interest. Hungarian Mangalica, likewise other native races, became uninteresting because of economic reasons or other characteristics, and were replaced by modern breeds. Its population decreased rapidly and reached a critical level. However, the exceptional taste of the meat, and the robustness and motherliness do support the propagation of this breed. Nevertheless, low prolificacy and marked seasonality remains a problem. The aim ofthe present study was to find possible implications of the physiological basis with regard to the low fecundity. Therefore, preovulatory follicular development and intrafollicular oocyte maturation of Mangalica and of Landrace gilts were compared. A total of 18 pubertal Blond and Swallow Belly Mangalica and 19 Landrace gilts (8.5 to 9 month of age, body weight of 120 to 125 kg) were used. Oestrus of gilts was synchronized by feeding Regumate®, follicular growth was stimulated by administering 1,000 IU PMSG 24 h after the last Regumate® feeding and the LH peak simulated with 750 IU hCG 80 h after PMSG. Cumulus-oocyte-complexes (COCs) were recovered 34 h after hCG by endoscopic Ovum Pick Up. Follicular fluids from follicles per ovary were pooled and the morphology of COCs was determined. COCs were classified as compact, expanded or denuded. Thereafter, COCs were prepared for evaluation of nuclear configuration. Based on their nuclear status the oocytes were classified as 1) immature – germinal vesicle (GV), with diplotene chromatin; 2) meiosis resumed – G V breakdown, diakinesis, M-I to A-I; or 3) mature – T-I and M-II. The average number (+SD) of preovulatory follicles was 6.8 + 1.4 in Mangalica and 19.6 ± 6.6 in Landrace gilts (p<0.05). Differences were obtained conceming the morphology of recovered COCs between breeds. The per cent of oocytes with compact cumulus was higher in Mangalica compared to Landrace gilts (31 vs. 16 %) but less oocytes possess expanded cumulus (62 vs. 78 %, p<0.05). The meiotic configuration of oocytes was unlike between Mangalica and Landrace gilts. The rate of oocytes with mature chromatin configuration (Telophase I /Metaphase II) was higher (27 vs. 62 %, p<0.05) in Landrace sows. It is assumed that both diminished follicular development and protracted intrafollicular oocyte maturation may be involved in low fecundity in Mangalica.

scholarly journals Genetic biomarkers in the VEGF pathway predicting response to anti-VEGF therapy in age-related macular degeneration

2019 ◽  
Vol 4 (1) ◽  
pp. e000273
Author(s):  
Irina Balikova ◽  
Laurence Postelmans ◽  
Brigitte Pasteels ◽  
Pascale Coquelet ◽  
Janet Catherine ◽  
...  
Keyword(s):  
Visual Acuity ◽  
Macular Degeneration ◽  
Treatment Response ◽  
Multiple Testing ◽  
Age Related Macular Degeneration ◽  
Patient Specific ◽  
Multiple Testing Correction ◽  
Anti Vegf ◽  
Age Related ◽  
Vegf Pathway

ObjectiveAge-related macular degeneration (ARMD) is a leading cause of visual impairment. Intravitreal injections of anti-vascular endothelial growth factor (VEGF) are the standard treatment for wet ARMD. There is however, variability in patient responses, suggesting patient-specific factors influencing drug efficacy. We tested whether single nucleotide polymorphisms (SNPs) in genes encoding VEGF pathway members contribute to therapy response.Methods and analysisA retrospective cohort of 281 European wet ARMD patients treated with anti-VEGF was genotyped for 138 tagging SNPs in the VEGF pathway. Per patient, we collected best corrected visual acuity at baseline, after three loading injections and at 12 months. We also registered the injection number and changes in retinal morphology after three loading injections (central foveal thickness (CFT), intraretinal cysts and serous neuroepithelium detachment). Changes in CFT after 3 months were our primary outcome measure. Association of SNPs to response was assessed by binomial logistic regression. Replication was attempted by associating visual acuity changes to genotypes in an independent Japanese cohort.ResultsAssociation with treatment response was detected for seven SNPs, including in FLT4 (rs55667289: OR=0.746, 95% CI 0.63 to 0.88, p=0.0005) and KDR (rs7691507: OR=1.056, 95% CI 1.02 to 1.10, p=0.005; and rs2305945: OR=0.963, 95% CI 0.93 to 1.00, p=0.0472). Only association with rs55667289 in FLT4 survived multiple testing correction. This SNP was unavailable for testing in the replication cohort. Of six SNPs tested for replication, one was significant although not after multiple testing correction.ConclusionIdentifying genetic variants that define treatment response can help to develop individualised therapeutic approaches for wet ARMD patients and may point towards new targets in non-responders.

scholarly journals 2dFDR: a new approach to confounder adjustment substantially increases detection power in omics association studies

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Sangyoon Yi ◽  
Xianyang Zhang ◽  
Lu Yang ◽  
Jinyan Huang ◽  
Yuanhang Liu ◽  
...  
Keyword(s):  
Multiple Testing ◽  
Statistical Power ◽  
Association Studies ◽  
Control Procedure ◽  
Multiple Testing Correction ◽  
New Approach ◽  
False Discovery ◽  
Traditional Procedure ◽  
Extensive Evaluation ◽  
Confounder Adjustment

AbstractOne challenge facing omics association studies is the loss of statistical power when adjusting for confounders and multiple testing. The traditional statistical procedure involves fitting a confounder-adjusted regression model for each omics feature, followed by multiple testing correction. Here we show that the traditional procedure is not optimal and present a new approach, 2dFDR, a two-dimensional false discovery rate control procedure, for powerful confounder adjustment in multiple testing. Through extensive evaluation, we demonstrate that 2dFDR is more powerful than the traditional procedure, and in the presence of strong confounding and weak signals, the power improvement could be more than 100%.

Induction of DNA replication in germinal vesicles and in nuclei formed in maturing mouse oocytes by 6-DMAP treatment

Zygote ◽  
1997 ◽  
Vol 5 (3) ◽  
pp. 213-217 ◽  
Author(s):  
J. Fulka ◽  
N.L. First ◽  
C. Lee ◽  
J. Fulka ◽  
R.M. Moor
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Germinal Vesicle ◽  
The Other ◽  
Immature Oocytes ◽  
Mouse Oocytes ◽  
Germinal Vesicles ◽  
Immature Mouse ◽  
Condensed Dna ◽  
Metaphase Ii Oocytes

SummaryImmature mouse oocytes (germinal vesicle stage, GV), oocytes at different stages during maturation (prometaphase to anaphase I) and matured oocytes (metaphase II arrested) were cultured in 6-dimethylaminopurine (6-DMAP)-supplemented medium also containing bromodeoxyuridine for the assessment of DNA replication in these cells. Immature oocytes remained arrested at the GV stage and DNA replication was never detected in them. On the other hand, oocytes at the prometaphase to anaphase-telophase I stages responded to 6-DMAP treatment by forming nuclei which synthesised DNA. Mature (metaphase II) oocytes did not respond to 6-DMAP and their chromatin remained condensed. DNA synthesis could even be induced in GV-staged oocytes, but only when they were fused to freshly activated oocytes and incubated in 6-DMAP-supplemented medium.

scholarly journals NRG1 Genetic Variant Influences the Efficacy of Androgen-Deprivation Therapy in Men with Prostate Cancer

Biomedicines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 528
Author(s):  
Shu-Pin Huang ◽  
Yei-Tsung Chen ◽  
Lih-Chyang Chen ◽  
Cheng-Hsueh Lee ◽  
Chao-Yuan Huang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Androgen Deprivation Therapy ◽  
Tyrosine Kinases ◽  
Multiple Testing ◽  
Cox Regression ◽  
Progression Free Survival ◽  
Androgen Deprivation ◽  
Nucleotide Polymorphisms ◽  
Multiple Testing Correction ◽  
Cancer Specific Survival

Neuregulins (NRGs) activate receptor tyrosine kinases of the ErbB family, and play essential roles in the proliferation, survival, and differentiation of normal and malignant tissue cells. We hypothesized that genetic variants of NRG signalling pathway genes may influence treatment outcomes in prostate cancer. To test this hypothesis, we performed a comprehensive analysis to evaluate the associations of 459 single-nucleotide polymorphisms in 19 NRG pathway genes with cancer-specific survival (CSS), overall survival (OS), and progression-free survival (PFS) in 630 patients with prostate cancer receiving androgen-deprivation therapy (ADT). After multivariate Cox regression and multiple testing correction, we found that NRG1 rs144160282 C > T is significantly associated with worsening CSS, OS, and PFS during ADT. Further analysis showed that low expression of NRG1 is closely related to prostate cancer, as indicated by a high Gleason score, an advanced stage, and a shorter PFS rate. Meta-analysis of 16 gene expression datasets of 1,081 prostate cancer samples and 294 adjacent normal samples indicate lower NRG1 expression in the former compared with the latter (p < 0.001). These results suggest that NRG1 rs144160282 might be a prognostic predictor of the efficacy of ADT. Further studies are required to confirm the significance of NRG1 as a biomarker and therapeutic target for prostate cancer.

scholarly journals Case-control study of adverse childhood experiences and multiple sclerosis risk and clinical outcomes

PLoS ONE ◽  
2022 ◽  
Vol 17 (1) ◽  
pp. e0262093
Author(s):  
Mary K. Horton ◽  
Shannon McCurdy ◽  
Xiaorong Shao ◽  
Kalliope Bellesis ◽  
Terrence Chinn ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Clinical Outcomes ◽  
Adverse Childhood Experiences ◽  
Multiple Testing ◽  
Multiple Testing Correction ◽  
Latent Factors ◽  
Childhood Experiences ◽  
Adverse Childhood ◽  
Rich Data ◽  
Control Study

Background Adverse childhood experiences (ACEs) are linked to numerous health conditions but understudied in multiple sclerosis (MS). This study’s objective was to test for the association between ACEs and MS risk and several clinical outcomes. Methods We used a sample of adult, non-Hispanic MS cases (n = 1422) and controls (n = 1185) from Northern California. Eighteen ACEs were assessed including parent divorce, parent death, and abuse. Outcomes included MS risk, age of MS onset, Multiple Sclerosis Severity Scale score, and use of a walking aid. Logistic and linear regression estimated odds ratios (ORs) (and beta coefficients) and 95% confidence intervals (CIs) for ACEs operationalized as any/none, counts, individual events, and latent factors/patterns. Results Overall, more MS cases experienced ≥1 ACE compared to controls (54.5% and 53.8%, respectively). After adjusting for sex, birthyear, and race, this small difference was attenuated (OR = 1.01, 95% CI: 0.87, 1.18). There were no trends of increasing or decreasing odds of MS across ACE count categories. Consistent associations between individual ACEs between ages 0–10 and 11–20 years and MS risk were not detected. Factor analysis identified five latent ACE factors, but their associations with MS risk were approximately null. Age of MS onset and other clinical outcomes were not associated with ACEs after multiple testing correction. Conclusion Despite rich data and multiple approaches to operationalizing ACEs, no consistent and statistically significant effects were observed between ACEs with MS. This highlights the challenges of studying sensitive, retrospective events among adults that occurred decades before data collection.

scholarly journals Genome-wide meta-analysis of depression identifies 102 independent variants and highlights the importance of the prefrontal brain regions

2018 ◽  
Author(s):  
David M. Howard ◽  
Mark J. Adams ◽  
Toni-Kim Clarke ◽  
Jonathan D. Hafferty ◽  
Jude Gibson ◽  
...  
Keyword(s):  
Multiple Testing ◽  
Drug Repositioning ◽  
Association Studies ◽  
Meta Analysis ◽  
Enrichment Analysis ◽  
Brain Regions ◽  
Genome Wide Association Studies ◽  
Multiple Testing Correction ◽  
Synaptic Structure ◽  
Genome Wide

AbstractMajor depression is a debilitating psychiatric illness that is typically associated with low mood, anhedonia and a range of comorbidities. Depression has a heritable component that has remained difficult to elucidate with current sample sizes due to the polygenic nature of the disorder. To maximise sample size, we meta-analysed data on 807,553 individuals (246,363 cases and 561,190 controls) from the three largest genome-wide association studies of depression. We identified 102 independent variants, 269 genes, and 15 gene-sets associated with depression, including both genes and gene-pathways associated with synaptic structure and neurotransmission. Further evidence of the importance of prefrontal brain regions in depression was provided by an enrichment analysis. In an independent replication sample of 1,306,354 individuals (414,055 cases and 892,299 controls), 87 of the 102 associated variants were significant following multiple testing correction. Based on the putative genes associated with depression this work also highlights several potential drug repositioning opportunities. These findings advance our understanding of the complex genetic architecture of depression and provide several future avenues for understanding aetiology and developing new treatment approaches.

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