Role of Intercellular Coupling on Chromatin Changes Transcriptional Activity and Meiotic Competence Acquisition During Bovine Oocyte Growth In Vitro.

2009 ◽  
Vol 81 (Suppl_1) ◽  
pp. 282-282
Author(s):  
Federica M. Franciosi ◽  
Valentina Lodde ◽  
Silvia Modina ◽  
Irene Tessaro ◽  
Alberto M. Luciano
Keyword(s):  
Transcriptional Activity ◽  
Bovine Oocyte ◽  
Intercellular Coupling ◽  
Oocyte Growth ◽  
Competence Acquisition ◽  
Meiotic Competence

scholarly journals Zinc supports transcription and improves meiotic competence of growing bovine oocytes

Reproduction ◽  
2020 ◽  
Vol 159 (6) ◽  
pp. 679-691
Author(s):  
Valentina Lodde ◽  
Rodrigo Garcia Barros ◽  
Priscila Chediek Dall’Acqua ◽  
Cecilia Dieci ◽  
Claude Robert ◽  
...  
Keyword(s):  
Culture Medium ◽  
Zinc Supplementation ◽  
Oocyte Growth ◽  
Zinc Chelator ◽  
Bovine Oocytes ◽  
Meiotic Competence ◽  
Medium Supplementation

In the last years, many studies focused on the understanding of the possible role of zinc in the control of mammalian oogenesis, mainly on oocyte maturation and fertilization. However, little is known about the role of zinc at earlier stages, when the growing oocyte is actively transcribing molecules that will regulate and sustain subsequent stages of oocyte and embryonic development. In this study, we used the bovine model to gain insights into the possible involvement of zinc in oocyte development. We first mined the EmbryoGENE transcriptomic dataset, which revealed that several zinc transporters and methallothionein are impacted by physiological conditions throughout the final phase of oocyte growth and differentiation. We then observed that zinc supplementation during in vitro culture of growing oocytes is beneficial to the acquisition of meiotic competence when subsequently subjected to standard in vitro maturation. Furthermore, we tested the hypothesis that zinc supplementation might support transcription in growing oocytes. This hypothesis was indirectly confirmed by the experimental evidence that the content of labile zinc in the oocyte decreases when a major drop in transcription occurs in vivo. Accordingly, we observed that zinc sequestration with a zinc chelator rapidly reduced global transcription in growing oocytes, which was reversed by zinc supplementation in the culture medium. Finally, zinc supplementation impacted the chromatin state by reducing the level of global DNA methylation, which is consistent with the increased transcription. In conclusion, our study suggests that altering zinc availability by culture-medium supplementation supports global transcription, ultimately enhancing meiotic competence.

scholarly journals The role of the 5′-flanking sequence of a human tRNAGlu gene in modulation of its transcriptional activity in vitro

1990 ◽  
Vol 272 (3) ◽  
pp. 797-803 ◽  
Author(s):  
E S Gonos ◽  
J P Goddard
Keyword(s):  
Transcriptional Activity ◽  
Potential Model ◽  
Trna Gene ◽  
Deletion Mutants ◽  
Wild Type ◽  
Flanking Sequence ◽  
Cell Extracts ◽  
Transcription In Vitro

The role of a tRNA-like structure within the 5′-flanking sequence of a human tRNA(Glu) gene in the modulation of its transcription in vitro by HeLa cell extracts has been investigated using several deletion mutants of a recombinant of the gene which lacked part or all of the tRNA-like structure. The transcriptional efficiency of four mutants was the same as that of the wild-type recombinant, two mutants had decreased transcriptional efficiency, one was more efficient, and one, lacking part of the 5′ intragenic control region, was inactive. Correlation of the transcriptional efficiencies with the position and the size of the 5′-flanking sequence that was deleted indicated that the tRNA-like structure may be deleted without loss of transcriptional efficiency. Current models for the modulation of tRNA gene transcription by the 5′-flanking sequence are assessed in the light of the results obtained, and a potential model is presented.

Role of Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mice in vitro

Zygote ◽  
2013 ◽  
Vol 22 (4) ◽  
pp. 513-520 ◽  
Author(s):  
I. Bahena ◽  
E. Xu ◽  
M. Betancourt ◽  
E. Casas ◽  
Y. Ducolomb ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Differentiation ◽  
Embryonic Stem Cells ◽  
Germ Cell ◽  
Embryonic Stem ◽  
Oocyte Growth ◽  
Germ Cell Differentiation ◽  
Rnai Technology

SummaryIn a previous study, we have identified a set of conserved spermatogenic genes whose expression is restricted to testis and ovary and that are developmentally regulated. One of these genes, the transcription factor Mael, has been reported to play an essential role in mouse spermatogenesis. Nevertheless, the role of Mael in mouse oogenesis has not been defined. In order to analyse the role of Mael in mouse oogenesis, the expression of this gene was blocked during early oogenesis in mouse in vitro using RNAi technology. In addition, the role of Mael during differentiation of embryonic stem cells (ESC) into germ cells in vitro was analysed. Results show that downregulation of Mael by a specific short interfering RNA disrupted fetal oocyte growth and differentiation in fetal ovary explants in culture and the expression of several germ-cell markers in ESC during their differentiation. These results suggest that there is an important role for Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mouse in vitro.

scholarly journals Tingkat Kematangan Inti Oosit Sapi Setelah 24 Jam Presevasi Ovarium

2015 ◽  
Vol 15 (2) ◽  
pp. 72-78
Author(s):  
Rini Widyastuti ◽  
Siti Darodjah Rasad
Keyword(s):  
Phosphate Buffer ◽  
Bovine Oocyte ◽  
Oocyte Maturity ◽  
Phosphate Buffer Saline ◽  
Bovine Oocytes ◽  
Meiotic Competence

(Nuclear maturity of bovine oocyte after 24 hours ovary preservation)ABSTRACT. The objective of the research was to investigate  their meiotic competence or nuclear maturity of bovine oocytes maturity in vitro after 24 hour preservation on 5°C. Oocytes  were collected by slicing the ovaries in modified phosphate buffer saline (m-PBS). Selected cumulus-oocyte complexes (COCs) homogenous ooplasm  were cultured in maturity medium at 38°C in humidified atmosphere of 5% CO2 incubator. After 24 hours, oocytes stained for nuclear maturity’s evaluation. The proportion of oocytes at metaphase II (MII) was significantly difference (P 0.05) on oocytes that 24 hours preservated   (44.21 ± 3.04%) vs oocytes from fresh ovary (73.97 ± 9.32%) (P0.05). These results indicated that 24 hour’s preservation  bovine’s ovary on 50C  cause  decreases of nuclear oocyte maturity.

Theca cells can support bovine oocyte growth in vitro without the addition of steroid hormones

2020 ◽  
Vol 142 ◽  
pp. 41-47 ◽  
Author(s):  
Yinghua Yang ◽  
Chihiro Kanno ◽  
Kenichiro Sakaguchi ◽  
Seiji Katagiri ◽  
Yojiro Yanagawa ◽  
...  
Keyword(s):  
Steroid Hormones ◽  
Bovine Oocyte ◽  
Oocyte Growth ◽  
Theca Cells

scholarly journals FKBP4 integrates FKBP4/Hsp90/IKK with FKBP4/Hsp70/RelA complex to promote lung adenocarcinoma progression via IKK/NF-κB signaling

2021 ◽  
Vol 12 (6) ◽  
Author(s):  
Shuai Zong ◽  
Yulian Jiao ◽  
Xin Liu ◽  
Wenli Mu ◽  
Xiaotian Yuan ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Transcriptional Activity ◽  
Nuclear Translocation ◽  
Critical Role ◽  
The Family ◽  
Ikk Complex ◽  
Breast And Prostate Cancer ◽  
Insight Into

AbstractFKBP4 belongs to the family of immunophilins, which serve as a regulator for steroid receptor activity. Thus, FKBP4 has been recognized to play a critical role in several hormone-dependent cancers, including breast and prostate cancer. However, there is still no research to address the role of FKBP4 on lung adenocarcinoma (LUAD) progression. We found that FKBP4 expression was elevated in LUAD samples and predicted significantly shorter overall survival based on TCGA and our cohort of LUAD patients. Furthermore, FKBP4 robustly increased the proliferation, metastasis, and invasion of LUAD in vitro and vivo. Mechanistic studies revealed the interaction between FKBP4 and IKK kinase complex. We found that FKBP4 potentiated IKK kinase activity by interacting with Hsp90 and IKK subunits and promoting Hsp90/IKK association. Also, FKBP4 promotes the binding of IKKγ to IKKβ, which supported the facilitation role in IKK complex assembly. We further identified that FKBP4 TPR domains are essential for FKBP4/IKK interaction since its association with Hsp90 is required. In addition, FKBP4 PPIase domains are involved in FKBP4/IKKγ interaction. Interestingly, the association between FKBP4 and Hsp70/RelA favors the transport of RelA toward the nucleus. Collectively, FKBP4 integrates FKBP4/Hsp90/IKK with FKBP4/Hsp70/RelA complex to potentiate the transcriptional activity and nuclear translocation of NF-κB, thereby promoting LUAD progression. Our findings suggest that FKBP4 may function as a prognostic biomarker of LUAD and provide a newly mechanistic insight into modulating IKK/NF-κB signaling.

280 EFFECTS OF HOT SEASON ON BOVINE OOCYTE QUALITY: HOW TO BYPASS THE POOR OOCYTE QUALITY DURING THIS SEASON?

2015 ◽  
Vol 27 (1) ◽  
pp. 229
Author(s):  
L. Boccia ◽  
E. Iacono ◽  
B. Rossi ◽  
B. Merlo
Keyword(s):  
Degradation Products ◽  
Germinal Vesicle ◽  
Oocyte Quality ◽  
Female Reproductive Tract ◽  
Bovine Oocyte ◽  
Germinal Vesicle Breakdown ◽  
Serum Replacement ◽  
Hot Season ◽  
Meiotic Competence

Many authors attribute the decline of reproductive activity in summer to the heat stress, a multifactorial problem in which hyperthermia affects cellular function in various tissues of the female reproductive tract (Hansen et al. 2001; De Rensis et al. 2003). In particular, the combination of high temperatures and high humidity for a long period causes a reduced blood flow to uterus, oviducts, and ovaries, leading to a rise in the concentration of the degradation products of cellular activity. Therefore, the aim of this work was to elucidate the negative effect of the hot season on bovine oocyte quality and evaluate the influence of different factors on the acquisition of meiotic competence. In particular, meiotic competence of bovine oocytes recovered from animals housed at 44°28′00″ N, 11°26′00″ E during spring (March, 4–13°C) and summer (June, 16–27°C) was evaluated. Likewise, in summer the effect of an antioxidant, myo-inositol, the use of serum replacement (SR), and the use of oocytes recovered from cycling heifers (16–18 months) as compared to cows (>24 months) were tested. A total of 1346 abattoir-derived oocytes, equally divided for different experimental groups (over 6 replicates), were in vitro matured in TCM 199 supplemented with EGF (25 ng mL–1), IGF1 (100 ng mL–1), ITS supplement, pFSH-LH (0.1 IU each), and 10% FBS. Myoinositol was added at a concentration of 0, 15, 30, and 50 mM, while 10% SR was used alternatively to FBS. At the end of maturation period (20–22 h), oocytes were denuded and stained with 10 μg mL–1 of Hoechst 33342 at room temperature in the dark. After 15 min they were mounted on glass slides for evaluation of nuclear status using a Nikon Eclipse E400 microscope equipped with fluorescence filters. Nuclear configurations were classified as (a) germinal vesicle (GV), (b) germinal vesicle breakdown (GVBD), (c) metaphase I (M-I), (d) metaphase II (M-II), and (e) degenerated (DEG). Data are expressed as mean ± s.e.m. and were analysed by ANOVA (IBM SPSS Statistics) considering significance at P < 0.05. Oocyte quality of summer oocytes was significantly lower than spring counterparts as result of a higher rate of DEG (8.2 ± 0.6 v. 0.7 ± 0.6) and GV (5.4 ± 0.3 v. 0.4 ± 0.4, respectively; P < 0.05). Myo-inositol supplementation in IVM medium did not significantly affect either oocyte quality or meiotic competence in the hot season, such as the use of SR. When the oocytes were collected from cycling heifers ovaries during summer, the recovery rate of COC/ovary was significantly higher as compared to cows (4.5 v. 2.0), and a lower rate of DEG (1.8 ± 0.2; 8.2 ± 0.6) and GVBD (0.9 ± 0.6; 6.1 ± 0.3) was found (P < 0.05), even if the rate of GV (22.4 ± 0.1 v. 5.4 ± 0.3) was higher (P < 0.05) compared with cow. In conclusion, the hot season negatively affects oocyte quality, myo-inositol does not affect nuclear maturation, and SR can be used alternatively to FBS. The lower age of oocyte donor positively influenced the number of recoverable oocyte and degeneration rate.

Abstract 34: Statins Directly Activate Erk5 and Reduce Endothelial Dysfunction and Acute Allograft Rejection

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Yuichiro Takei ◽  
Nhat-Tu Le ◽  
Hakjoo Lee ◽  
Kyung-Sun Heo ◽  
Cheryl Hurley ◽  
...  
Keyword(s):  
Allograft Rejection ◽  
High Throughput Screening ◽  
Transcriptional Activity ◽  
Vascular Reactivity ◽  
Pleiotropic Effects ◽  
Protective Effects ◽  
Acute Allograft Rejection

Rationale: The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or statins, are principal therapeutic agents for the treatment of hypercholesteremia. Statins, however, appear to also exert cholesterol-independent pleiotropic effects, such as improvement of endothelial (EC) function, stabilization of fibrous plaques, and decrease vascular inflammation. It is now well established that statins are beneficial in the prevention of cardiovascular disease and also widely used for suppressing cardiac allograft rejection. Previously, we have reported anti-inflammatory effect of ERK5 kinase in ECs. Methods and Results: In this study, we screened small molecules that activate ERK5 using high throughput screening, and identified statins as strong activators of the transcriptional activity of ERK5. In particular, we have found that pitavastatin increases ERK5 transcriptional activity, KLF2 promoter activity, and eNOS mRNA expression in ECs. These effects are abolished by the depletion of ERK5, but not its direct upstream kinase, MEK5. In addition, pitavastatin directly and dose-dependently activates ERK5 kinase activity in an in vitro kinase reaction assay, suggesting that ERK5 is a direct target of this statin. To examine the functional role of EC ERK5 activation by the statin in vivo, we utilized inducible endothelial ERK5 knock out (EC-ERK5-KO) mice and evaluated the effect of pitavastatin on EC function and acute allograft rejection. Depletion of ERK5 in ECs resulted in significant EC inflammation and dysfunction in vivo. Although pitavastatin reduced leukocyte rolling and vascular reactivity in mesebteruc microvessels of diabetic mice and prolonged allograft survival in a full allomismatch combination model, these protective effects were lost in EC-ERK5-KO mice. Conclusion: These data suggest the crucial role of ERK5 in pleiotropic effects of statins on EC dysfunction and allograft rejection in vivo.

Growth in vitro and acquisition of meiotic competence after the cryopreservation of isolated mouse primary ovarian follicles

1991 ◽  
Vol 3 (5) ◽  
pp. 593 ◽  
Author(s):  
J Carroll ◽  
DG Whittingham ◽  
MJ Wood
Keyword(s):  
Subsequent Development ◽  
Collagen Gels ◽  
Freezing And Thawing ◽  
Oocyte Growth ◽  
Large Numbers ◽  
Mean Diameter ◽  
Old Mice ◽  
Meiotic Competence

The growth and acquisition of meiotic competence of oocytes from fresh and frozen-thawed primary follicles collected from 10-day-old mice was compared during culture in collagen gels for 12 days. The oocytes contained in primary follicles have a mean diameter of about 48 microns and do not resume meiosis without further growth and development. During the 12-day culture period the mean diameter of the oocytes increased to over 60 microns. The oocytes were capable of resuming meiosis when isolated from the gel and cultured in the absence of follicular cells in a manner similar to that observed in vivo. Freezing and thawing did not affect oocyte growth or the ability to resume meiosis; this demonstrates the possibility of storing large numbers of female gametes for subsequent development.

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