scholarly journals Effects of Porcine Immature Oocyte Vitrification on Actin Microfilament Distribution and Chromatin Integrity During Early Embryo Development in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Alma López ◽  
Yvonne Ducolomb ◽  
Eduardo Casas ◽  
Socorro Retana-Márquez ◽  
Miguel Betancourt ◽  
...  
Keyword(s):  
Embryo Development ◽  
Early Embryo ◽  
Actin Microfilaments ◽  
Cell Protection ◽  
Developmental Potential ◽  
Early Embryo Development ◽  
Immature Oocytes ◽  
Actin Microfilament ◽  
Development Rates ◽  
Chromatin Integrity

Vitrification is mainly used to cryopreserve female gametes. This technique allows maintaining cell viability, functionality, and developmental potential at low temperatures into liquid nitrogen at −196°C. For this, the addition of cryoprotectant agents, which are substances that provide cell protection during cooling and warming, is required. However, they have been reported to be toxic, reducing oocyte viability, maturation, fertilization, and embryo development, possibly by altering cell cytoskeleton structure and chromatin. Previous studies have evaluated the effects of vitrification in the germinal vesicle, metaphase II oocytes, zygotes, and blastocysts, but the knowledge of its impact on their further embryo development is limited. Other studies have evaluated the role of actin microfilaments and chromatin, based on the fertilization and embryo development rates obtained, but not the direct evaluation of these structures in embryos produced from vitrified immature oocytes. Therefore, this study was designed to evaluate how the vitrification of porcine immature oocytes affects early embryo development by the evaluation of actin microfilament distribution and chromatin integrity. Results demonstrate that the damage generated by the vitrification of immature oocytes affects viability, maturation, and the distribution of actin microfilaments and chromatin integrity, observed in early embryos. Therefore, it is suggested that vitrification could affect oocyte repair mechanisms in those structures, being one of the mechanisms that explain the low embryo development rates after vitrification.

scholarly journals 012.Metabolic determinants of implantation success and programming long term viability in embryos

2004 ◽  
Vol 16 (9) ◽  
pp. 12
Author(s):  
J. G. Thompson ◽  
K. L. Kind
Keyword(s):  
Amino Acid ◽  
Embryo Development ◽  
Expression Patterns ◽  
Early Embryo ◽  
Bovine Embryo ◽  
Acid Uptake ◽  
Developmental Potential ◽  
Early Embryo Development ◽  
Causal Pathways ◽  
Implantation Success

It has long been recognised that energy substrate supply and metabolism are key determinants of early embryo development during in vitro culture. Recently it has been revealed that exposure to suboptimal metabolic environments during early embryo development can 'programme' subsequent development, leading to perturbed fetal development. For example, amino acid uptake profiles during early cleavage have been found to predict subsequent embryo development and potentially implantation success. However, the by-product of amino acid metabolism, ammonium, has also been found to significantly alter development, possibly through perturbed methylation of imprinted genes. Our own work has focussed on the role of oxygen availability and subsequent embryo development. Somatic cells respond to changing oxygen concentration by altering intracellular REDOX state (the balance between oxidative and reductive power within a cell), which in turn can alter transcription via REDOX-sensitive transcription factor activity. Furthermore, oxygen is known to have direct effects on transcriptional activity via the hypoxia-inducible factors (HIFs), transcription factors whose stability and DNA-binding activity are directly regulated by pO2, in particular under hypoxic conditions. Using a mouse model, we have demonstrated that reducing pO2 from 50�mmHg to 15�mmHg during the compaction and blastulation periods alone significantly alters expression patterns of oxygen-sensitive genes (such as glucose transporters), without significantly altering developmental progression to the blastocyst stage. Following transfer, embryos cultured under 15�mmHg O2, despite similar implantation rates, produced fewer viable and lighter fetuses than in vivo-derived control embryos or those cultured in either atmospheric or 50�mmHg pO2. This demonstrates that mouse embryos are sensitive to changes in their metabolic state during the post compaction period and that operating through causal pathways, the environment during this period of development can significantly affect subsequent developmental potential. Ironically, bovine embryo development appears to benefit under a low O2 concentration. Furthermore, HIF protein stability appears to differ between the two species, which may be the underlying cause for the differences in gene expression and developmental competence.

134 THE DISTRIBUTION OF CYTOSKELETON AND CHROMATIN CONFIGURATIONS DURING EARLY EMBRYO DEVELOPMENT OF SWAMP BUFFALO (BUBALUS BUBALIS)

2010 ◽  
Vol 22 (1) ◽  
pp. 226
Author(s):  
M. Techakumphu ◽  
V. Chankitisakul ◽  
K. Thaseephoo ◽  
T. Tharasanit
Keyword(s):  
Embryo Development ◽  
Early Embryo ◽  
Actin Microfilaments ◽  
Dense Network ◽  
Early Embryo Development ◽  
Vitro Fertilization ◽  
Sperm Penetration ◽  
Swamp Buffalo ◽  
Sperm Aster

Microtubules and actin microfilaments have been demonstratedto be actively involved with fertilization and early embryo development. The objective of this study was to examine the redistribution of cytoskeleton and chromatin configurations in swamp buffalo oocytes through the initial cleavage event after in vitro fertilization. Sperm penetration was analysed at 6 h post IVF in 63 oocytes (3 replicates), whereas the chronology embryonic development in terms of the redistribution of cell cytoskeleton and chromatin configurations was studied in a total of 462 oocytes (7 to 8 replicates) at 12, 18, 24, 30, and 48 h after IVF. The oocytes were matured in vitro for 22 h. Then, IVF was performed as described previously (Totey et al. 1993). After fertilization, presumptive zygotes and embryos were fixed at various times (6, 12, 18, 24, 30, and 48 h) to examine spermatozoa penetration, redistribution of the cytoskeleton (microtubules and actin filaments), and chromatin configurations using epifluorescent microscopy. Staining was undertaken with wheat germ agglutinin to visualize the zona pellucida, monoclonal-α-tubulin-TRIT C to show the microtubules, 488 phalloidin to identify microfilaments, and DAPI to label the chromatin. At 6 h after fertilization, sperm penetration was observed in 44.4% of examined oocytes. At 12 h post IVF, maternal chromosomes of fertilized oocytes progressed to the second meiotic division and formed the female pronucleus simultaneously with the decondensation of paternal chromosomes. A dense network of microtubules was observed radiating from the base of the decondensing sperm head (referred to as sperm aster) At 18 h post IVF, the sperm chromatins became the male pronucleus. Simultaneously, the sperm aster increased in size and filled the whole ooplasm. The syngamy of the male and female pronuclei was completed by 24 h post IVF, which was associated with a dense array of microtubules. Cell cleavage was observed by 30 h post IVF. This was apparently facilitated by a dense network of actin microfilaments that formed in the middle of the dividing embryo. These results indicated that microtubules and actin microfilaments undergo changes after fertilization consistent with a crucial role during fertilization in swamp buffalo. The centrosomal material was paternally inherited. This work was supported by TRF-MAG (MRG-WII515S056) and CHE-TRF Senior Research Fund (RTA5080010).

Minor Splicing Factors Zrsr1 and Zrsr2 Essential for Gametogenesis, Early Embryo Development and Conversion of Stem Cells into 2C-Like

2019 ◽  
Author(s):  
Isabel Gómez-Redondo ◽  
Priscila Ramos-Ibeas ◽  
Eva Pericuesta ◽  
Benjamín Planells ◽  
Raul Fernández-González ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryo Development ◽  
Early Embryo ◽  
Splicing Factors ◽  
Early Embryo Development

scholarly journals LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells

2021 ◽  
Author(s):  
Zhen Sun ◽  
Hua Yu ◽  
Jing Zhao ◽  
Tianyu Tan ◽  
Hongru Pan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryo Development ◽  
Pluripotent Stem Cells ◽  
Rna Binding ◽  
Stem Cell Differentiation ◽  
Early Embryo ◽  
Transcriptional Program ◽  
Cell Stage ◽  
Early Embryo Development ◽  
Nucleolar Stress

AbstractLIN28 is an RNA binding protein with important roles in early embryo development, stem cell differentiation/reprogramming, tumorigenesis and metabolism. Previous studies have focused mainly on its role in the cytosol where it interacts with Let-7 microRNA precursors or mRNAs, and few have addressed LIN28’s role within the nucleus. Here, we show that LIN28 displays dynamic temporal and spatial expression during murine embryo development. Maternal LIN28 expression drops upon exit from the 2-cell stage, and zygotic LIN28 protein is induced at the forming nucleolus during 4-cell to blastocyst stage development, to become dominantly expressed in the cytosol after implantation. In cultured pluripotent stem cells (PSCs), loss of LIN28 led to nucleolar stress and activation of a 2-cell/4-cell-like transcriptional program characterized by the expression of endogenous retrovirus genes. Mechanistically, LIN28 binds to small nucleolar RNAs and rRNA to maintain nucleolar integrity, and its loss leads to nucleolar phase separation defects, ribosomal stress and activation of P53 which in turn binds to and activates 2C transcription factor Dux. LIN28 also resides in a complex containing the nucleolar factor Nucleolin (NCL) and the transcriptional repressor TRIM28, and LIN28 loss leads to reduced occupancy of the NCL/TRIM28 complex on the Dux and rDNA loci, and thus de-repressed Dux and reduced rRNA expression. Lin28 knockout cells with nucleolar stress are more likely to assume a slowly cycling, translationally inert and anabolically inactive state, which is a part of previously unappreciated 2C-like transcriptional program. These findings elucidate novel roles for nucleolar LIN28 in PSCs, and a new mechanism linking 2C program and nucleolar functions in PSCs and early embryo development.

scholarly journals Geminin deletion in mouse oocytes results in impaired embryo development and reduced fertility

2016 ◽  
Vol 27 (5) ◽  
pp. 768-775 ◽  
Author(s):  
Xue-Shan Ma ◽  
Fei Lin ◽  
Zhong-Wei Wang ◽  
Meng-Wen Hu ◽  
Lin Huang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Primordial Follicle ◽  
Early Embryo ◽  
Cell Cycle Checkpoint ◽  
Low Fertility ◽  
Early Embryo Development ◽  
Damage Repair ◽  
Cycle Checkpoint ◽  
Oocyte Meiotic Maturation ◽  
Zygote Stage

Geminin controls proper centrosome duplication, cell division, and differentiation. We investigated the function of geminin in oogenesis, fertilization, and early embryo development by deleting the geminin gene in oocytes from the primordial follicle stage. Oocyte-specific disruption of geminin results in low fertility in mice. Even though there was no evident anomaly of oogenesis, oocyte meiotic maturation, natural ovulation, or fertilization, early embryo development and implantation were impaired. The fertilized eggs derived from mutant mice showed developmental delay, and many were blocked at the late zygote stage. Cdt1 protein was decreased, whereas Chk1 and H2AX phosphorylation was increased, in fertilized eggs after geminin depletion. Our results suggest that disruption of maternal geminin may decrease Cdt1 expression and cause DNA rereplication, which then activates the cell cycle checkpoint and DNA damage repair and thus impairs early embryo development.

scholarly journals The Potential Roles of the G1LEA and G3LEA Proteins in Early Embryo Development and in Response to Low Temperature and High Salinity in Artemia sinica

PLoS ONE ◽  
2016 ◽  
Vol 11 (9) ◽  
pp. e0162272 ◽  
Author(s):  
Wei Zhao ◽  
Feng Yao ◽  
Mengchen Zhang ◽  
Ting Jing ◽  
Shuang Zhang ◽  
...  
Keyword(s):  
Low Temperature ◽  
Embryo Development ◽  
High Salinity ◽  
Early Embryo ◽  
Artemia Sinica ◽  
Early Embryo Development

Early embryo development anomalies identified by time-lapse system: prevalence and impacting factors

2021 ◽  
Author(s):  
Xavier Ferraretto ◽  
Karima Hammas ◽  
Marie-Astrid Llabador ◽  
Solenne Gricourt ◽  
Julie Labrosse ◽  
...  
Keyword(s):  
Embryo Development ◽  
Time Lapse ◽  
Early Embryo ◽  
Early Embryo Development ◽  
Impacting Factors

Nutritional Status Impacts Epigenetic Regulation in Early Embryo Development: A Scoping Review

2021 ◽  
Author(s):  
Shuang Cai ◽  
Shuang Quan ◽  
Guangxin Yang ◽  
Meixia Chen ◽  
Qianhong Ye ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Epigenetic Regulation ◽  
Scoping Review ◽  
Epigenetic Modification ◽  
Reproductive Technologies ◽  
Early Embryo ◽  
Epigenetic Modifications ◽  
Nutrition Status ◽  
Early Embryo Development

ABSTRACTWith the increasing maternal age and the use of assisted reproductive technology in various countries worldwide, the influence of epigenetic modification on embryonic development is increasingly notable and prominent. Epigenetic modification disorders caused by various nutritional imbalance would cause embryonic development abnormalities and even have an indelible impact on health in adulthood. In this scoping review, we summarize the main epigenetic modifications in mammals and the synergies among different epigenetic modifications, especially DNA methylation, histone acetylation, and histone methylation. We performed an in-depth analysis of the regulation of various epigenetic modifications on mammals from zygote formation to cleavage stage and blastocyst stage, and reviewed the modifications of key sites and their potential molecular mechanisms. In addition, we discuss the effects of nutrition (protein, lipids, and one-carbon metabolism) on epigenetic modification in embryos and emphasize the importance of various nutrients in embryonic development and epigenetics during pregnancy. Failures in epigenetic regulation have been implicated in mammalian and human early embryo loss and disease. With the use of reproductive technologies, it is becoming even more important to establish developmentally competent embryos. Therefore, it is essential to evaluate the extent to which embryos are sensitive to these epigenetic modifications and nutrition status. Understanding the epigenetic regulation of early embryo development will help us make better use of reproductive technologies and nutrition regulation to improve reproductive health in mammals.

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