scholarly journals Effects of heat stress during in vitro maturation on cytoplasmic versus nuclear components of mouse oocytes

Reproduction ◽  
2009 ◽  
Vol 137 (2) ◽  
pp. 181-189 ◽  
Author(s):  
Jun-Zuo Wang ◽  
Hong-Shu Sui ◽  
De-Qiang Miao ◽  
Na Liu ◽  
Ping Zhou ◽  
...  
Keyword(s):  
Heat Stress ◽  
Similar Rate ◽  
Blastocyst Formation ◽  
Cortical Granules ◽  
Cell Stage ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Mouse Oocytes

The objectives of this study were to investigate the effect of heat stress duringin vitromaturation on the developmental potential of mouse oocytes and to determine whether the deleterious effect was on the nuclear or cytoplasmic component. While rates of oocyte nuclear maturation (development to the metaphase II stage) did not differ from 37 to 40 °C, rates for blastocyst formation decreased significantly as maturation temperature increased from 38.5 to 39 °C. Chromosome spindle exchange showed that while blastocyst formation did not differ when spindles maturedin vivoorin vitroat 37, 40 or 40.7 °C were transplanted intoin vivomatured cytoplasts, no blastocyst formation was observed whenin vivospindles were transferred into the 40 °C cytoplasts. While oocytes reconstructed between 37 °C ooplasts and 37 or 40 °C karyoplasts developed into 4-cell embryos at a similar rate, no oocytes reconstituted between 40 °C ooplasts and 37 °C spindles developed to the 4-cell stage. Immunofluorescence microscopy revealed impaired migration of cortical granules and mitochondria in oocytes matured at 40 °C compared with oocytes matured at 37 °C. A decreased glutathione/GSSG ratio was also observed in oocytes matured at 40 °C. While spindle assembling was normal and no MAD2 was activated in oocytes matured at 37 or 40 °C, spindle assembling was affected and MAD2 was activated in some of the oocytes matured at 40.7 °C. It is concluded that 1) oocyte cytoplasmic maturation is more susceptible to heat stress than nuclear maturation, and 2) cytoplasmic rather than nuclear components determine the pre-implantation developmental capacity of an oocyte.

scholarly journals Comparative Analyses of Single-Cell Transcriptomic Profiles between In Vitro Totipotent Blastomere-like Cells and In Vivo Early Mouse Embryonic Cells

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3111
Author(s):  
Po-Yu Lin ◽  
Denny Yang ◽  
Chi-Hsuan Chuang ◽  
Hsuan Lin ◽  
Wei-Ju Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Embryonic Cells ◽  
Cell Stage ◽  
Developmental Potential ◽  
Functional Features ◽  
Early Mouse ◽  
Extraembryonic Tissues

The developmental potential within pluripotent cells in the canonical model is restricted to embryonic tissues, whereas totipotent cells can differentiate into both embryonic and extraembryonic tissues. Currently, the ability to culture in vitro totipotent cells possessing molecular and functional features like those of an early embryo in vivo has been a challenge. Recently, it was reported that treatment with a single spliceosome inhibitor, pladienolide B (plaB), can successfully reprogram mouse pluripotent stem cells into totipotent blastomere-like cells (TBLCs) in vitro. The TBLCs exhibited totipotency transcriptionally and acquired expanded developmental potential with the ability to yield various embryonic and extraembryonic tissues that may be employed as novel mouse developmental cell models. However, it is disputed whether TBLCs are ‘true’ totipotent stem cells equivalent to in vivo two-cell stage embryos. To address this question, single-cell RNA sequencing was applied to TBLCs and cells from early mouse embryonic developmental stages and the data were integrated using canonical correlation analyses. Differential expression analyses were performed between TBLCs and multi-embryonic cell stages to identify differentially expressed genes. Remarkably, a subpopulation within the TBLCs population expressed a high level of the totipotent-related genes Zscan4s and displayed transcriptomic features similar to mouse two-cell stage embryonic cells. This study underscores the subtle differences between in vitro derived TBLCs and in vivo mouse early developmental cell stages at the single-cell transcriptomic level. Our study has identified a new experimental model for stem cell biology, namely ‘cluster 3’, as a subpopulation of TBLCs that can be molecularly defined as near totipotent cells.

Effect of trichostatin A on fertilization and embryo development during extended culture of mouse oocyte

Zygote ◽  
2011 ◽  
Vol 20 (1) ◽  
pp. 27-32 ◽  
Author(s):  
Byung Chul Jee ◽  
Jun Woo Jo ◽  
Jung Ryeol Lee ◽  
Chang Suk Suh ◽  
Seok Hyun Kim ◽  
...  
Keyword(s):  
Histone Deacetylase ◽  
Embryo Development ◽  
Trichostatin A ◽  
Formation Rate ◽  
Control Group ◽  
Blastocyst Formation ◽  
Developmental Potential ◽  
Extended Culture ◽  
Mouse Oocytes

SummaryWe performed this study to investigate the effect of histone deacetylase inhibition during extended culture of in vitro matured mouse oocytes. In vitro matured mouse (BDF1) oocytes were cultured in vitro for 6, 12, and 24 h, respectively, and then inseminated. During in vitro culture for 6 and 12 h, two doses of trichostatin A (TSA), a histone deacetylase inhibitor, were added (100 nM and 500 nM) to the culture medium and the oocytes were then inseminated. During the 24-h in vitro culture, two doses of TSA were added (100 nM and 500 nM) to the medium and the oocytes were activated with 10 mM SrCl2. After the 6-h culture, the fertilization rate was similar to that of the control group, but the blastocyst formation rate was significantly decreased. After the 12-h culture, both the fertilization and blastocyst formation rates were significantly decreased. After the 24-h culture, total fertilization failure occurred. In the oocytes cultured for 6 and 12 h, the fertilization and blastocyst formation rates did not differ between the TSA-supplemented and control groups. Although extended culture of the mouse oocytes significantly affected their fertilization and embryo development, TSA supplementation did not overcome their decreased developmental potential.

scholarly journals Influence of co-culture during maturation on the developmental potential of equine oocytes fertilized by intracytoplasmic sperm injection (ICSI)

Reproduction ◽  
2001 ◽  
pp. 925-932 ◽  
Author(s):  
X Li ◽  
LH Morris ◽  
WR Allen
Keyword(s):  
Epithelial Cells ◽  
Intracytoplasmic Sperm Injection ◽  
Fibroblast Cells ◽  
Cell Stage ◽  
Developmental Potential ◽  
Fetal Fibroblast ◽  
Nuclear Maturation ◽  
Fetal Fibroblasts ◽  
Oviduct Epithelial Cells

The influence of co-culture with either oviduct epithelial cells or fetal fibroblast cells on in vitro maturation of equine oocytes and their potential for development to blastocysts and fetuses after intracytoplasmic sperm injection (ICSI) was investigated. The oocytes were obtained from ovaries from abattoirs and were matured in vitro for 28-30 h in TCM-199 only, or in TCM-199 co-culture with oviduct epithelial cells or fetal fibroblast cells. Metaphase II oocytes were subjected to ICSI with an ionomycin-treated spermatozoon. The injected oocytes were cultured for 7-9 days in Dulbecco's modified Eagle's medium. Morphologically normal early blastocysts were transferred to the uteri of recipient mares. Nuclear maturation rates and the rates of cleavage to the two-cell stage for injected oocytes were similar in the groups of oocytes that were matured in TCM-199 (49 and 63%), in co-culture with oviduct epithelial cells (53 and 65%) or in co-culture with fetal fibroblasts (51 and 57%). There were no significant differences in the proportions of blastocysts that developed from the two-cell embryos derived from oocytes matured by co-culture with either oviduct epithelial cells (30%) or fetal fibroblasts (17%). However, significantly higher proportions of blastocysts were produced from both these co-culture groups than from the groups of oocytes matured in TCM-199 only (P < 0.05). Six of the blastocysts that had developed from oocytes co-cultured with oviduct epithelial cells were transferred into recipient mares and four pregnancies resulted. These results demonstrate a beneficial influence of co-culture with either oviduct epithelial cells or fetal fibroblasts for maturation of oocytes in vitro.

scholarly journals Full-term development of enucleated mouse oocytes fused with embryonic stem cells from different cell lines

Reproduction ◽  
2001 ◽  
pp. 729-733 ◽  
Author(s):  
T Amano ◽  
Y Kato ◽  
Y Tsunoda
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Lines ◽  
Formation Rate ◽  
Embryonic Stem ◽  
Clear Correlation ◽  
Developmental Potential ◽  
Mouse Oocytes

The developmental potential of enucleated mouse oocytes receiving embryonic stem cells from ten lines with either the same or different genetic backgrounds using the cell fusion method was examined in vitro and in vivo. The development of nuclear-transferred oocytes into blastocysts was high (34-88%). However, there was no clear correlation between development into blastocysts after nuclear transfer and the chimaera formation rate of embryonic stem cells. The development into live young was low (1-3%) in all cell lines and 14 of 19 young died shortly after birth. Most of the live young had morphological abnormalities. Of the five remaining mice, two died at days 23 and 30 after birth, but the other three mice are still active at days 359 (mouse 1) and 338 (mice 4 and 5) after birth, with normal fertility. However, the reasons for the abnormalities and postnatal death of embryonic stem cell-derived mice are unknown.

Relationship between bovine oocyte morphology and in vitro developmental potential

Zygote ◽  
2006 ◽  
Vol 14 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Masashi Nagano ◽  
Seiji Katagiri ◽  
Yoshiyuki Takahashi
Keyword(s):  
Small Diameter ◽  
Blastocyst Stage ◽  
Cortical Granules ◽  
Developmental Potential ◽  
Developmental Capacity ◽  
Sperm Penetration ◽  
Developmental Rates ◽  
Large Clusters

We investigated the relationship between the morphology of oocytes collected from small antral follicles and their developmental capacity. Immature oocytes were classified into seven groups and cultured in vitro for maturation (IVM), fertilization (IVF) and development to blastocysts (IVC). After IVF, sperm penetration and normal fertilization rates were higher in the oocytes whose cytoplasm appeared brown. The rate of polyspermy was highest in the oocytes whose cytoplasm was black. After IVC, the rates of cleavage and of development to the blastocyst stage were also higher in the brown oocytes. Although the oocytes with dark clusters in a pale cytoplasm showed lower cleavage rates, cleaved zygotes had high developmental rates the same as the oocytes with a brown cytoplasm. Transmission electron microscopy showed that the oocytes with a pale or black cytoplasm had organelles arranged differently from other oocytes before IVM. Most of the oocytes with a brown and homogeneous cytoplasm or small diameter had the characteristics of immature cytoplasm (large clusters of cortical granules) even after IVM. On the other hand, the brown oocytes with a dark zone at the periphery or with dark clusters showed the same arrangement of organelles as in vivo matured oocytes. The oocytes with a pale or black cytoplasm appeared to be degenerating and/or ageing. In conclusion, a dark ooplasm indicates an accumulation of lipids and good developmental potential, while a light-coloured ooplasm indicates a low density of organelles and poor developmental potential. A black ooplasm indicates ageing and low developmental potential.

215 INVESTIGATION OF A PREFERENTIALLY UPREGULATED GENE CLUSTER IN DAY 7 BOVINE EMBRYOS DERIVED FROM RNA SEQUENCING DATA

2013 ◽  
Vol 25 (1) ◽  
pp. 256 ◽  
Author(s):  
A. Al Naib ◽  
S. Mamo ◽  
P. Lonergan
Keyword(s):  
Rna Sequencing ◽  
Mature Oocyte ◽  
Blastocyst Stage ◽  
Blastocyst Formation ◽  
Sequencing Data ◽  
Cell Stage ◽  
Uterine Endometrium ◽  
Embryonic Origin

Successful establishment and maintenance of pregnancy requires optimum conceptus-maternal cross talk. Despite significant progress in our understanding of the temporal changes in the transcriptome of the uterine endometrium, we have only a rudimentary knowledge of the genes and pathways governing growth and development of the bovine conceptus. A recent RNA sequencing study from our group (Mamo et al. 2011 Biol. Reprod. 85, 1143–1151) described the global transcriptome profile of the bovine conceptus at 5 key stages of its pre- and peri-implantation growth (Days 7, 10, 13, 16, and 19) using RNA sequencing techniques. One cluster of genes (n = 1680 transcripts) was preferentially upregulated at Day 7 and subsequently downregulated, suggesting that these genes might be markers of blastocyst formation. The objective of this study was to characterise the pattern of expression of these genes before Day 7 (i.e. from the zygote to blastocyst stage). The list of genes was submitted to DAVID (Database for Annotation, Visualisation, and Integrated Discovery) to take advantage of available ontology information contained therein. The expression of 9 genes belonging to ontologies specifically related to embryo developmental (GINS1, TAF8, ESRRB, NCAPG2, SP1, XAB2, CDC2L1, MSX1, and AQP3) plus Na/K ATPase, a gene previously known to be involved in blastocoe formation, was studied by quantitative real-time PCR (QPCR) in 6 replicate pools of 5 embryos produced by maturation, fertilization, and embryo culture in vitro. Stages studies included immature and mature oocyte, zygote, 2- cell, 4-cell, 8-cell, 16-cell, morula, blastocyst, and hatched blastocyst. In addition, in vivo derived Day 13 and Day 16 embryos were included as controls to confirm down-regulation after Day 7. Data were analysed using the GLM procedure of SAS. The QPCR expression data supported the RNA Seq data in that expression of all transcripts was downregulated after the blastocyst stage. Expression before the blastocyst stage was characterised by 1 of 3 broad patterns: (1) the expression was of maternal origin where the expression was very high up to 8-cell stage and decreased subsequently (MSX1), (2) the expression was of embryonic origin being low up to the 8-cell stage and increasing thereafter (TAF8, ESRRB, AQP3, and Na/K ATPase), or (3) static or decreased expression from oocyte to the maternal-zygotic transition followed by increased expression from the 16-cell stage (GINS1, NCAPG2, SP1, XAB2, and CDC2L1). In conclusion, the genes identified in this cluster, despite having different patterns of expression before the blastocyst stage, may represent markers of blastocyst formation in cattle given their downregulation subsequently. Supported by Science Foundation Ireland (07/SRC/B1156).

Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 501-507 ◽  
Author(s):  
M.H. Nasr-Esfahani ◽  
J.R. Aitken ◽  
M.H. Johnson
Keyword(s):  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Cell Stage ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Individual Mouse ◽  
The Relationship ◽  
Developmental Block

We describe a fluorimetric method for measuring the level of H2O2 in individual mouse oocytes and early embryos. Levels of H2O2 are low but detectable in unfertilized oocytes recovered freshly from the female reproductive tract. The levels in early cleaving embryos (1-cell to 8-cell stages) immediately after recovery from the female tract seem to be slightly higher the later the stage examined. However, when embryos are cultured in vitro from the 1-cell or early 2-cell stage, H2O2 levels rise when the embryos reach the mid-2-cell stage and remain elevated until they enter the early 4-cell stage. No equivalent elevation of H2O2 is seen during the transition from 1-cell to 2-cell or from 4-cell to 8-cell stages. Embryos that are able to develop successfully in vitro, as well as those that show a developmental block at the 2-cell stage on culture in vitro, both show this rise in H2O2 levels after in vitro culture. The relationship between the rise in H2O2 and the ‘2-cell block’ to development is discussed.

P–204 Dynamics of cavitation as a potential non-invasive predictor of mammalian blastocyst quality

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
E Kosyl ◽  
A Ajduk
Keyword(s):  
Mouse Model ◽  
Mouse Embryo ◽  
Embryo Quality ◽  
Mammalian Species ◽  
Cavity Formation ◽  
Developmental Potential ◽  
Mouse Oocytes ◽  
Maternal Aging

Abstract Study question We wished to investigate whether dynamics of cavity formation can be used in embryo quality assessment. Summary answer Dynamics of mouse embryo cavitation reflects to certain extent blastocysts’ developmental capabilities. It can be potentially used as a biomarker of mammalian embryo quality. What is known already During cavity expansion blastocyst pulsates, i.e. changes its volume in an oscillatory way. Recent studies performed on a mouse model have shown, that dynamics of cavitation, biomechanical properties of the trophectoderm (TE) and embryo size are intertwined. Presence or absence of blastocyst contractions has been linked to particular parameters related to positive outcome of the in vitro fertilization procedures, but the data on influence of contractions on human embryos’ developmental capabilities is often contradictory. Moreover, mostly in those studies only strong contractions (leading to a high volume loss) have been taken into consideration. Study design, size, duration We tested how postovulatory (in vitro or in vivo) or maternal aging of mouse oocytes affects dynamics of cavity formation and expansion in the resulting embryos (n = 27, n = 26 and n = 30, respectively). Furthermore, we also analyzed almost 100 mouse blastocysts in order to correlate dynamics of their cavitation with their ability to form correct outgrowths (in vitro model of implantation). Participants/materials, setting, methods Mouse oocytes subjected to postovulatory (either in vivo or in vitro) or maternal aging were fertilized in vitro. Dynamics of cavity formation and expansion was assessed by time-lapse imaging; equatorial images were taken every 10 minutes. Blastocyst area was measured over time and compared to the outcome from control embryos. In another set of experiments, after the filming mouse blastocysts were cultured for additional 4 days to test their ability to form outgrowths. Main results and the role of chance We noticed, that mouse embryos which represent limited developmental potential (obtained from either postovulatory or maternally aged oocytes) and blastocysts developed from freshly fertilized young females’ oocytes differ in terms of some parameters related to dynamics of cavitation, e.g. time of the initiation of cavity formation, frequency of contractions or mean loss of blastocyst’s area during contraction. We observed that embryos obtained from oocytes subjected to maternal or postovulatory aging have distinct dynamics of cavitation. Moreover, we noticed slightly different effect on particular parameters related to cavitation between in vivo and in vitro version of postovulatory ageing. We also showed that blastocysts, which are unable to create proper outgrowths (i.e. too small or without epiblast cells), differ from embryos that differentiate into correct outgrowths in terms of certain parameters of cavitation dynamics. Our data indicates, that dynamics of cavity formation and expansion might be related to developmental potential of mouse embryo. Limitations, reasons for caution Further studies with extended group size and testing embryos’ ability to implant in vivo are required to confirm our results. Moreover, we examined dynamics of cavitation only in a mouse model, so additional studies performed on other mammalian species are needed. Wider implications of the findings: Our data proves, that dynamics of embryo cavitation reflects, to certain extent, developmental capabilities of mouse blastocysts. Therefore, it is possible that it can be a biomarker of embryo quality (in combination with parameters provided by other methods or solely) of other mammalian species, including humans. Trial registration number Not applicable

Sperm-derived factors enhance the in vitro developmental potential of haploid parthenotes

Zygote ◽  
2017 ◽  
Vol 25 (6) ◽  
pp. 697-710 ◽  
Author(s):  
Ramya Nair ◽  
Shahin Aboobacker ◽  
Srinivas Mutalik ◽  
Guruprasad Kalthur ◽  
Satish Kumar Adiga
Keyword(s):  
Strontium Chloride ◽  
Cortical Granules ◽  
Cell Stage ◽  
Developmental Potential ◽  
Activation Rate ◽  
Paternal Factors ◽  
Artificial Activation ◽  
Cortical Distribution

SummaryParthenotes are characterized by poor in vitro developmental potential either due to the ploidy status or the absence of paternal factors. In the present study, we demonstrate the beneficial role of sperm-derived factors (SDF) on the in vitro development of mouse parthenotes. Mature (MII) oocytes collected from superovulated Swiss albino mice were activated using strontium chloride (SrCl2) in the presence or absence of various concentrations of SDF in M16 medium. The presence of SDF in activation medium did not have any significant influence on the activation rate. However, a significant increase in the developmental potential of the embryos and increased blastocyst rate (P < 0.01) was observed at 50 µg/ml concentration. Furthermore, the activated oocytes from this group exhibited early cleavage and cortical distribution of cortical granules that was similar to that of normally fertilized zygotes. Culturing 2-cell stage parthenotes in the presence of SDF significantly improved the developmental potential (P < 0.05) indicating that they also play a significant role in embryo development. In conclusion, artificial activation of oocytes with SDF can improve the developmental potential of parthenotes in vitro.

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