The movement of amino acids into rabbit oviductal fluid

Leese, Henry J.; Aldridge, Sue; Jeffries, Kate S.

doi:10.1530/jrf.0.0560623

Cited by 23 publications

(11 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our results however suggest that the G1+ medium may not be entirely adapted to development when the embryo is ‘preparing’ for EGA. This cannot be explained by the lack of carbohydrates, as the concentrations of glucose, lactate, pyruvate and citrate are very similar in both G1+ and Global, but may be related to the absence of essential amino acids in G1+ medium (Morbeck et al ., 2014), while those are present in rabbit oviductal fluid (Leese et al ., 1979). Conversely, the Global medium appears adapted to the EGA developmental period but may not be suited to the later developmental stages (from 16/32-cell).…”

Section: Discussionmentioning

confidence: 99%

Assessment of ‘one-step’ versus ‘sequential’ embryo culture conditions through embryonic genome methylation and hydroxymethylation changes

et al. 2016

View full text Add to dashboard Cite

STUDY QUESTIONIn comparison to in vivo development, how do different conditions of in vitro culture (‘one step’ versus ‘sequential medium’) impact DNA methylation and hydroxymethylation in preimplantation embryos?SUMMARY ANSWERUsing rabbit as a model, we show that DNA methylation and hydroxymethylation are both affected by in vitro culture of preimplantation embryos and the effect observed depends on the culture medium used.WHAT IS KNOWN ALREADYCorrect regulation of DNA methylation is essential for embryonic development and DNA hydroxymethylation appears more and more to be a key player. Modifications of the environment of early embryos are known to have long term effects on adult phenotypes and health; these probably rely on epigenetic alterations.STUDY DESIGN SIZE, DURATIONThe study design we used is both cross sectional (control versus treatment) and longitudinal (time-course). Each individual in vivo experiment used embryos flushed from the donor at the 2-, 4-, 8-, 16- or morula stage. Each stage was analyzed in at least two independent experiments. Each individual in vitro experiment used embryos flushed from donors at the 1-cell stage (19 h post-coïtum) which were then cultured in parallel in the two tested media until the 2-, 4-, 8- 16-cell or morula stages. Each stage was analyzed in at least three independent experiments. In both the in vivo and in vitro experiments, 4-cell stage embryos were always included as an internal reference.PARTICIPANTS/MATERIALS, SETTING, METHODSImmunofluorescence with antibodies specific for 5-methylcytosine (5meC) and 5-hydroxymethylcytosine (5hmeC) was used to quantify DNA methylation and hydroxymethylation levels in preimplantation embryos. We assessed the expression of DNA methyltransferases (DNMT), of ten eleven translocation (TET) dioxigenases and of two endogenous retroviral sequences (ERV) using RT-qPCR, since the expression of endogenous retroviral sequences is known to be regulated by DNA methylation. Three repeats were first done for all stages; then three additional repetitions were performed for those stages showing differences or tendencies toward differences between the different conditions in the first round of quantification.MAIN RESULTS AND THE ROLE OF CHANCEThe kinetics of DNA methylation and hydroxymethylation were modified in in vitro cultured embryos, and the observed differences depended on the type of medium used. These differences were statistically significant. In addition, the expression of TET1 and TET2 was significantly reduced in post-embryonic genome activation (EGA) embryos after in vitro culture in both tested conditions. Finally, the expression of two retroviral sequences was analyzed and found to be significantly affected by in vitro culture.LIMITATIONS REASONS FOR CAUTIONOur study remains mostly descriptive as no direct link can be established between the epigenetic changes observed and the expression changes in both effectors and targets of the studied epigenetic modifications. The results we obtained suggest that gene expression could be a...

show abstract

Section: Discussionmentioning

confidence: 99%

Assessment of ‘one-step’ versus ‘sequential’ embryo culture conditions through embryonic genome methylation and hydroxymethylation changes

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Leese & Jeffries (1977) examined the appearance of endogenous glucose in fluid recirculated through the lumen of the rabbit oviduct for 4 h in vivo, and obtained evidence that the glucose moved by facilitated diffusion. Similar experiments were carried out on pyruvate, lactate and amino acid transport Leese, Aldridge & Jeffries, 1979). This approach has now been used to compare the transport of glucose pyruvate and lactate into the lumen of the rabbit ampulla and isthmus.…”

Section: Introductionmentioning

confidence: 98%

Studies on the Movement of Glucose, Pyruvate and Lactate Into the Ampulla and Isthmus of the Rabbit Oviduct

Leese

1983

Exp Physiol

Self Cite

View full text Add to dashboard Cite

SUMMARYThe appearance of glucose, pyruvate and lactate in fluid recirculated through the lumen of the ampulla and isthmus of the rabbit oviduct has been followed for 2 h in vivo. Each nutrient appeared in the lumen of the ampulla at approximately 1 8 times its rate in the lumen of the isthmus. The circumference of the mucosa lining the two regions was measured together with the distribution of ciliated and secretory cells. The surface area of the mucosa lining the ampulla was 343 mm2, and that lining the isthmus, 191 mm2. It is tentatively suggested that the capacity of the ampulla and isthmus to transport small molecules is a function of their respective mucosal surface areas.

show abstract

“…Female reproductive tract fluid contains high concentrations of certain amino acids [1,2], suggesting that they may play a role in preimplantation development. This is supported by the observation that embryos have the ability to take up amino acids [3][4][5][6][7], which contribute to large endogenous pools at preimplantation stages [2,[8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Effects of Glutamine and Taurine on Preimplantation Development and Cleavage of Mouse Embryos in Vitro1

Devreker¹,

Hardy²

1997

Biology of Reproduction

View full text Add to dashboard Cite

Glutamine and taurine are reported to improve the development of mouse embryos in vitro, and glutamine is a component in CZB and KSOM, media optimized to overcome the 2-cell block in outbred strains of mice. Here we have studied the development of embryos from F1 hybrid female mice in CZB and KSOM, from the zygote to the blastocyst stage, in the presence and absence of 1 mM glutamine and/or 5 mM taurine. Blastocyst formation, rate of development, number of trophectoderm and inner cell mass cells, mitosis, and cell death were examined. Glutamine had no effect on the proportion of embryos developing to the blastocyst stage, but it significantly reduced cell number at the blastocyst stage. The trophectoderm was the lineage most affected, showing increased levels of cell death. Changing the media after 48 h, to remove any possible buildup of toxic metabolites, did not reverse the inhibition by glutamine. Taurine had no effect on blastocyst development of cell number. Development in glutamine-free KSOM (KSOM-Gln) was compared to that in standard CZB and KSOM with glutamine and M16. Early cleavage was slower in M16, but by Day 5 over 84% of embryos had developed to the blastocyst stage in all media, with a similar cell number in M16, CZB, and KSOM. Cell numbers in blastocysts cultured in KSOM-Gln were significantly higher than in the other media and approached those found in vivo.

show abstract

The movement of amino acids into rabbit oviductal fluid

Cited by 23 publications

References 4 publications

Assessment of ‘one-step’ versus ‘sequential’ embryo culture conditions through embryonic genome methylation and hydroxymethylation changes

Assessment of ‘one-step’ versus ‘sequential’ embryo culture conditions through embryonic genome methylation and hydroxymethylation changes

Studies on the Movement of Glucose, Pyruvate and Lactate Into the Ampulla and Isthmus of the Rabbit Oviduct

Effects of Glutamine and Taurine on Preimplantation Development and Cleavage of Mouse Embryos in Vitro1

Contact Info

Product

Resources

About