scholarly journals Intraoviductal Instillation of a Solution as an Effective Route for Manipulating Preimplantation Mammalian Embryos in vivo

2018 ◽  
Author(s):  
Masahiro Sato ◽  
Masato Ohtsuka ◽  
Shingo Nakamura
Keyword(s):  
Mammalian Embryos

scholarly journals The Xenoestrogen Bisphenol A Inhibits Postembryonic Vertebrate Development by Antagonizing Gene Regulation by Thyroid Hormone

Endocrinology ◽  
2009 ◽  
Vol 150 (6) ◽  
pp. 2964-2973 ◽  
Author(s):  
Rachel A. Heimeier ◽  
Biswajit Das ◽  
Daniel R. Buchholz ◽  
Yun-Bo Shi
Keyword(s):  
Bisphenol A ◽  
Estrogenic Activity ◽  
Sex Differentiation ◽  
Inhibitory Effect ◽  
Vertebrate Development ◽  
Developmental Context ◽  
Mammalian Embryos ◽  
Intestinal Remodeling

Bisphenol A (BPA), a chemical widely used to manufacture plastics, is estrogenic and capable of disrupting sex differentiation. However, recent in vitro studies have shown that BPA can also antagonize T3 activation of the T3 receptor. The difficulty in studying uterus-enclosed mammalian embryos has hampered the analysis on the direct effects of BPA during vertebrate development. This study proposed to identify critical T3 pathways that may be disrupted by BPA based on molecular analysis in vivo. Because amphibian metamorphosis requires T3 and encompasses the postembryonic period in mammals when T3 action is most critical, we used this unique model for studying the effect of BPA on T3-dependent vertebrate development at both the morphological and molecular levels. After 4 d of exposure, BPA inhibited T3-induced intestinal remodeling in premetamorphic Xenopus laevis tadpoles. Importantly, microarray analysis revealed that BPA antagonized the regulation of most T3-response genes, thereby explaining the inhibitory effect of BPA on metamorphosis. Surprisingly, most of the genes affected by BPA in the presence of T3 were T3-response genes, suggesting that BPA predominantly affected T3-signaling pathways during metamorphosis. Our finding that this endocrine disruptor, well known for its estrogenic activity in vitro, functions to inhibit T3 pathways to affect vertebrate development in vivo and thus not only provides a mechanism for the likely deleterious effects of BPA on human development but also demonstrates the importance of studying endocrine disruption in a developmental context in vivo.

51 ARTIFICIAL DORMANCY OF BOVINE EMBRYOS FOR A MAXIMUM OF 7 DAYS USING A SIMPLE MEDIUM

2014 ◽  
Vol 26 (1) ◽  
pp. 139 ◽  
Author(s):  
K. Tsuchiya ◽  
A. Ideta ◽  
Y. Nishimiya ◽  
S. Tsuda ◽  
Y. Aoyagi
Keyword(s):  
Pregnancy Rate ◽  
Storage Period ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Hypothermic Preservation ◽  
Mammalian Embryos ◽  
Simple Medium ◽  
Hypothermic Storage

The worldwide pregnancy rate using cryopreserved mammalian embryos has not improved over the past 2 decades, probably because the freeze-thawing processes cause significant damage. Therefore, it is now relevant to examine the feasibility of short-term non-freezing preservation, and whether this could be applied to embryos that have high vitality and are to be transferred into recipients within several days. We introduce here an artificial dormancy fluid that can extend the hypothermic storage period of bovine embryos for a maximum of 7 days. First, to examine the effect of different basal media and the optimal concentration of fetal bovine serum (FBS) for hypothermic preservation, bovine blastocysts produced in vitro were stored at 4°C in a plastic ministraw in 1 of the following 3 media: PBS, medium 199, or Leibovitz L15 with various amount of FBS (0, 5, 20, 50, or 100%) for 3 days. Second, to examine the effect of Good's buffers, bovine embryos produced in vivo (morula to blastocyst stages) were stored at 4°C in a plastic ministraw in medium 199 plus 50% FBS supplemented with various Good's buffers [HEPES, TES, piperazine-N,N′-bis(2-ethanesulfonic acid) (PIPES), MOPS, and 4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid (EPPS)] for 7 days. Following hypothermic preservation, the chilled embryos were squeezed out of the straw into PBS and washed 3 times in the same medium. Subsequently, the embryos were cultured in CR1aa medium supplemented with 5% FBS for 48 h at 38.5°C under 5% CO2 in air with high humidity. The viability rate of the embryos was assessed at the end of the culture period. Finally, to observe the pregnancy rate of chilled embryos, 32 embryos produced in vivo were stored at 4°C for 7 days in medium 199 plus 50% FBS supplemented with HEPES. Following hypothermic preservation, the chilled embryos were transferred into recipient heifers (1 embryo per recipient). Pregnancy was determined by real-time B-mode ultrasonography (Convex scanner HS-1500, Honda electronics Co. Ltd, Toyohashi, Japan) on Day 60 of gestation. Data were analysed using the chi-squared test. The viability rate of the embryos after hypothermic storage for 3 days was significantly increased for medium 199 plus 50% FBS [27/30 (90%)] compared with PBS [18/30 (60%)] or Leibovitz L15 [15/30 (50%)] plus 50% FBS (P < 0.05). Chilled embryos stored for 7 days in medium 199 plus 50% FBS supplemented with HEPES had much higher survival than embryos stored in the same medium with other Good's buffers. The pregnancy rate of the chilled embryos stored for 7 days was extremely high [24/32 (75%)] and normal live calves were delivered at term. In conclusion, maintaining artificial dormancy of bovine embryos for 7 days using a simple medium appears to be feasible. This is the first documented success of storing chilled mammalian embryos in a viable state for 7 days. To be of practical value, bovine embryo preservation at hypothermic temperatures must be able to maintain viability for periods longer than 7 days. This work was supported by the Program for Promotion of Basic and Applied Research for Innovations in Bio-Oriented Industry.

scholarly journals Composing the Early Embryonic Microenvironment: Physiology and Regulation of Oviductal Secretions

2019 ◽  
Vol 21 (1) ◽  
pp. 223 ◽  
Author(s):  
Marie Saint-Dizier ◽  
Jennifer Schoen ◽  
Shuai Chen ◽  
Charles Banliat ◽  
Pascal Mermillod
Keyword(s):  
Embryonic Development ◽  
Farm Animals ◽  
Meat Production ◽  
Mammalian Embryos ◽  
Production Techniques ◽  
Species Specific ◽  
Molecular Components ◽  
Oviductal Fluid

The oviductal fluid is the first environment experienced by mammalian embryos at the very beginning of life. However, it has long been believed that the oviductal environment was not essential for proper embryonic development. Successful establishment of in vitro embryo production techniques (which completely bypass the oviduct) have reinforced this idea. Yet, it became evident that in vitro produced embryos differ markedly from their in vivo counterparts, and these differences are associated with lower pregnancy outcomes and more health issues after birth. Nowadays, researchers consider the oviduct as the most suitable microenvironment for early embryonic development and a substantial effort is made to understand its dynamic, species-specific functions. In this review, we touch on the origin and molecular components of the oviductal fluid in mammals, where recent progress has been made thanks to the wider use of mass spectrometry techniques. Some of the factors and processes known to regulate oviductal secretions, including the embryo itself, as well as ovulation, insemination, endogenous and exogenous hormones, and metabolic and heat stress, are summarized. Special emphasis is laid on farm animals because, owing to the availability of sample material and the economic importance of fertility in livestock husbandry, a large part of the work on this topic has been carried out in domestic animals used for dairy and/or meat production.

scholarly journals Using the Amniotic Cavity of the Developing Chick Embryo for the In Vivo Culture of Early-Stage Mammalian Embryos ,

1989 ◽  
Vol 68 (12) ◽  
pp. 1695-1702 ◽  
Author(s):  
E.G. BLAKEWOOD ◽  
J.M. JAYNES ◽  
W.A. JOHNSON ◽  
R.A. GODKE
Keyword(s):  
Chick Embryo ◽  
Early Stage ◽  
Amniotic Cavity ◽  
In Vivo Culture ◽  
Mammalian Embryos

scholarly journals PRDM14 controls X-chromosomal and global epigenetic reprogramming of H3K27me3 in migrating mouse primordial germ cells

2019 ◽  
Author(s):  
Anna Mallol ◽  
Maria Guirola ◽  
Bernhard Payer
Keyword(s):  
X Chromosome ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Epigenetic Reprogramming ◽  
Spatiotemporal Resolution ◽  
Developmental Context ◽  
Inactive X Chromosome ◽  
A Genome ◽  
Mammalian Embryos

ABSTRACTIn order to prepare the genome for gametogenesis, primordial germ cells (PGCs) undergo extensive epigenetic reprogramming during migration towards the gonads in mammalian embryos. This includes changes on a genome-wide scale and additionally in females the remodeling of the inactive X-chromosome to enable X-chromosome reactivation (XCR). However, if global and X-chromosomal remodeling are related and which factors are important is unknown. Here we identify the germ cell determinant PR-domain containing protein 14 (PRDM14) as the first known factor that is instrumental for both global and X-chromosomal reprogramming in migrating mouse PGCs. We find that global upregulation of the repressive histone H3 lysine 27 trimethylation (H3K27me3) mark is PRDM14 dosage-dependent in PGCs of both sexes. When focusing on XCR, we observed that PRDM14 is required for removal of H3K27me3 from the inactive X-chromosome. Furthermore we show that global and X-chromosomal H3K27me3 reprogramming are functionally separable, despite their common regulation by PRDM14. Thereby we provide mechanistic insight and spatiotemporal resolution to the remodeling of the epigenome during mouse PGC migration and link epigenetic reprogramming to its developmental context in vivo.

scholarly journals In Vivo Imaging of Mammalian Embryos By NIR-I Photoacoustic Tomography and NIR-II Optical Coherence Tomography Using Gold Nanostars as Multifunctional Contrast Agents

2021 ◽  
Author(s):  
Sheng Zhang ◽  
Zhenyang Liu ◽  
Linlin Mao ◽  
Jian Wu ◽  
Di Zhang ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Embryonic Development ◽  
Contrast Agents ◽  
Structural Characteristic ◽  
Photoacoustic Tomography ◽  
Optical Coherence ◽  
Foetal Development ◽  
Gold Nanostars ◽  
Mammalian Embryos

Abstract Background High resolution, strong contrast and multimodality visualization of live mammalian embryo is an important requirement for studying foetal development. Photoacoustic Tomography (PAT) and Optical Coherence Tomography (OCT) are two advanced imaging modalities that has been utilized for embryonic imaging. However, high contrast, multiscale and deep tissue visualization of live embryos remains challenging. Results Here, we demonstrate the use of gold nanostars (GNS) as multimodality contrast agents for the visualization and differentiation of embryos in vivo using NIR-I PAT and NIR-II OCT. We perform NIR-I PAT imaging to confirm in vivo GNS accumulation in the foetuses, and then use a customized NIR-II OCT system to further reveal deep, contrast-enhanced micro features of freshly harvested embryos. We investigate two different GNS administration pathways, i.e. intravenous and intravaginal injection, and significant enhancement of signal, image contrast, and imaging depth are achieved for both PAT and OCT. Conclusions These findings prove that PAT-OCT bi-modal imaging with GNS enhancement provides more accurate structural characteristic of live mammalian embryos, and thus reveal its potential for embryonic development visualization and early abnormality examination. These findings prove that PAT-OCT bi-modal imaging with GNS enhancement provides more accurate structural characteristic of live mammalian embryos, and thus reveal its potential for embryonic development visualization and early abnormality examination.

Expression of the T85A mutant of zebrafish aquaporin 3b improves post-thaw survival of cryopreserved early mammalian embryos

Zygote ◽  
2016 ◽  
Vol 24 (6) ◽  
pp. 839-847 ◽  
Author(s):  
Sylvia J. Bedford-Guaus ◽  
François Chauvigné ◽  
Eva Mejía-Ramírez ◽  
Mercè Martí ◽  
Antoni Ventura-Rubio ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Survival Rates ◽  
Ectopic Expression ◽  
Membrane Expression ◽  
Cell Stage ◽  
C Terminus ◽  
Mammalian Embryos ◽  
Mouse Zygotes ◽  
Toxic Concentrations

SummaryWhile vitrification has become the method of choice for preservation of human oocytes and embryos, cryopreservation of complex tissues and of large yolk-containing cells, remains largely unsuccessful. One critical step in such instances is appropriate permeation while avoiding potentially toxic concentrations of cryoprotectants. Permeation of water and small non-charged solutes, such as those used as cryoprotectants, occurs largely through membrane channel proteins termed aquaporins (AQPs). Substitution of a Thr by an Ala residue in the pore-forming motif of the zebrafish (Dario rerio) Aqp3b paralog resulted in a mutant (DrAqp3b-T85A) that when expressed in Xenopus or porcine oocytes increased their permeability to ethylene glycol at pH 7.5 and 8.5. The main objective of this study was to test whether ectopic expression of DrAqp3b-T85A also conferred higher resistance to cryoinjury. For this, DrAqp3b-T85A + eGFP (reporter) cRNA, or eGFP cRNA alone, was microinjected into in vivo fertilized 1-cell mouse zygotes. Following culture to the 2-cell stage, appropriate membrane expression of DrAqp3b-T85A was confirmed by immunofluorescence microscopy using a primary specific antibody directed against the C-terminus of DrAqp3b. Microinjected 2-cell embryos were then cryopreserved using a fast-freezing rate and low concentration (1.5 M) of ethylene glycol in order to highlight any benefits from DrAqp3b-T85A expression. Notably, post-thaw survival rates were higher (P<0.05) for T85A–eGFP-injected than for -uninjected or eGFP-injected embryos (73±7.3 vs. 28±7.3 or 14±6.7, respectively). We propose that ectopic expression of mutant AQPs may provide an avenue to improve cryopreservation results of large cells and tissues in which current vitrification protocols yield low survival.

The Effects of AZT and DDI on Pre- and Postimplantation Mammalian Embryos: An In Vivo and In Vitro Study

1992 ◽  
Vol 8 (5) ◽  
pp. 639-649 ◽  
Author(s):  
EMMANUEL SIEH ◽  
M. LUISA COLUZZI ◽  
M. GABRIELLA CUSELLA DE ANGELIS ◽  
ANTONIO MEZZOGIORNO ◽  
MARCO FLORIDIA ◽  
...  
Keyword(s):  
In Vitro Study ◽  
Vitro Study ◽  
Mammalian Embryos

scholarly journals Cytoplasmic streaming in Drosophila oocytes varies with kinesin activity and correlates with the microtubule cytoskeleton architecture

2012 ◽  
Vol 109 (38) ◽  
pp. 15109-15114 ◽  
Author(s):  
Sujoy Ganguly ◽  
Lucy S. Williams ◽  
Isabel M. Palacios ◽  
Raymond E. Goldstein
Keyword(s):  
Cytoplasmic Streaming ◽  
The Body ◽  
Meiotic Spindle ◽  
Microtubule Cytoskeleton ◽  
Developmental Potential ◽  
Image Velocimetry ◽  
Mammalian Embryos ◽  
Combined Action

Cells can localize molecules asymmetrically through the combined action of cytoplasmic streaming, which circulates their fluid contents, and specific anchoring mechanisms. Streaming also contributes to the distribution of nutrients and organelles such as chloroplasts in plants, the asymmetric position of the meiotic spindle in mammalian embryos, and the developmental potential of the zygote, yet little is known quantitatively about the relationship between streaming and the motor activity which drives it. Here we use Particle Image Velocimetry to quantify the statistical properties of Kinesin-dependent streaming during mid-oogenesis in Drosophila. We find that streaming can be used to detect subtle changes in Kinesin activity and that the flows reflect the architecture of the microtubule cytoskeleton. Furthermore, based on characterization of the rheology of the cytoplasm in vivo, we establish estimates of the number of Kinesins required to drive the observed streaming. Using this in vivo data as the basis of a model for transport, we suggest that the disordered character of transport at mid-oogenesis, as revealed by streaming, is an important component of the localization dynamics of the body plan determinant oskar mRNA.

scholarly journals NMR spectroscopy of a single mammalian embryo

2021 ◽  
Author(s):  
Giulia Sivelli ◽  
Gaurasundar Marc Conley ◽  
Carolina Herrera ◽  
Kathryn Marable ◽  
Kyle Rodriguez ◽  
...  
Keyword(s):  
Building Blocks ◽  
Resolving Power ◽  
In Vivo Studies ◽  
Small Samples ◽  
Single Chip ◽  
Mammalian Embryo ◽  
Non Invasive ◽  
Spectral Peaks ◽  
Mammalian Embryos

The resolving power, chemical sensitivity and non-invasive nature of NMR has made it an established technique for in vivo studies of large organisms both for research and clinical applications. These features would clearly be beneficial at the nanoliter scale (nL), typical of early development of mammalian embryos, microtissues and organoids, the scale where the building blocks of complex organisms could be observed. However, the handling of such small samples (about 100 micrometers) and sensitivity issues have prevented the widespread adoption of NMR. Recently we have shown how these limitations can be overcome with ultra-compact single-chip probes. In this article we show that such probes have sufficient sensitivity to detect and resolve NMR signals from individual bovine pre-implantation embryos. In less than 1 hour these spherical samples of just 130-190 micrometers produce distinct spectral peaks, largely originating from lipids contained inside them. We further observe how the spectral features, namely the peak intensities, vary from one sample to another despite their optical and morphological similarities.

Sign in / Sign up

Export Citation Format

Share Document