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Influence of dietary protein restriction on ovulation, fertilization rates and pre-implantation embryonic development in mice.

Eggs were recovered from 153 adult female mice fed either a normal (27% protein) or a protein-deficient (8% protein) diet from day 15 (Group I) or 30 (Group II) before mating until sacrifice. The females were mated with fertile males and sacrificed 48, 72, 96 or 120 hours after observation of vaginal plugs. The study was performed with females that ovulated spontaneously and with females induced to ovulate by injection of pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. Without gonadotrophin treatment, significantly fewer eggs were recovered from malnourished than from normal females. However, after hormonal treatment, the number of eggs ovulated by malnourished and normal females increased similarly. About 50% of the eggs recovered from malnourished females in both groups I and II were cleaved whereas 90% of the eggs recovered from well-nourished females were cleaved. The development of the cleaved eggs from protein deficient females was delayed and asynchronous in comparison with that of eggs from normal females. Embryos recovered from malnourished females also were retarded in differentiation of morulae to blastocysts. It is concluded that a protein-restricted diet adversely affects the fertilization of eggs as well as their early development and differentiation.

Animals

A long noncoding RNA with enhancer-like function in pig zygotic genome activation.

The zygotic genome activation (ZGA) is crucial for the development of pre-implantation embryos. Long noncoding RNAs (lncRNAs) play significant roles in many biological processes, but the study on their role in the early embryonic development of pigs is limited. In this study, we identify lncFKBPL as an enhancer-type lncRNA essential for pig embryo development. lncFKBPL is expressed from the 4-cell stage to the morula stage in pig embryos, and interference with lncFKBPL leads to a developmental arrest at the 8-cell stage. Mechanistic investigations uncover that lncFKBPL is able to bind to MED8, thereby mediating enhancer activity and regulating FKBPL expression. Additionally, FKBPL interacts with the molecular chaperone protein HSP90AA1, stabilizing CDK9 and boosting its protein-level expression. Elevated CDK9 levels enhance Pol II phosphorylation, facilitating ZGA. Our findings illuminate the role of lncFKBPL as an enhancer lncRNA in pig ZGA regulation and early embryo development, providing a foundation for further exploration in this area.

Animals

[Morphologic and histochemical characteristics of mammalian embryos during the pre-implantation period of development].

Submicroscopic embryo organization during preimplantation development was analyzed and its carbohydrate metabolism at these stages was studied. In the process of investigation the following regularities were revealed: progressive rearrangement of the nucleus (from compact to reticular); alterations in cytoplasmic structures (in multivesicular bodies, in autophagal vacuoles, in mitochondria, in granular cytoplasmic network, in lamellar structures); complication of intracellular interrelations (by means of microvilli and median bodies at early stages and during subsequent formation of cellular processes, contacts after "thorn--net" pattern, dense, fissure-like contacts and desmosomes). By means of cytochemical and electron histochemical methods, glycogen distribution in embryos at different stages of development was elucidated, and tendency to its decrease with the progress of the development was noted, with its least contents being at the stage of blastocyst.

Animals

Foetal and placental growth in the mouse after pre-implantation development in vitro under oxygen concentrations of 5 and 20%.

Blastocysts which developed from two-cell mouse embryos in culture tubes containing an atmosphere with 20% oxygen had approximately 20% fewer blastomeres than blastocysts which developed under an oxygen concentration of 5%. When these smaller blastocysts were transferred to the uteri of pseudopregnant foster mothers, the foetuses developing were as viable as those developing from blastocysts cultured under 5% oxygen, indicating their ability to regulate for a lower blastomere number by at least day 17 of development. The transfer operation itself had no adverse effect on foetal or placental growth. However, culture of blastocysts in vitro did depress foetal though not placental growth, suggesting that the inner cell mass is more susceptible than the trophectoderm to culture in vitro. Foetal but not placental growth was lower following the transfer of blastocysts to a day-3 rather than a day-4 uterus. Four cases of placental fusion were found. In one case, the foetuses were contained within the same embryonic sac and may have been twins.

Animals

The ontogenic characteristics of lactate dehydrogenase isozymes in mammaliam pre-implantation ova.

The onotgenic characteristics of lactate dehydrogenase (LDH) isozymes in mammalian pre-implantation ova have been reviewed. Evidence has been provided that the ova of mice and other mammalian species contain enzyme activity in a masked form, and display turnover processes which possess distinctive characteristics by comparison with those in adult tissues. Also, the extraordinary high levels of LDH in the extracellular secretion of the mammaliam oviduct have been commented on, along with the influence of reproductive hormones on the activity and type of this enzyme. In addition, attention has been drawn to the unique characteristics of the oval micro-evironment, and the influence which such factors may exert on the realization of enzyme phenotype during early mammalian development.

Animals

[Effect of aminopterin on pre-implantation rat embryos cultivated outside the body].

The rat embryos at the stage of 8 blastomeres were explanted in the Biggers' medium with 20% of blood serum from the rats obtained different doses of aminopterin. The marked delay of cleavage rate, morphological anomalies of blastocysts, selective damaging effect of aminopterin upon the inner cells mass (embryoblast) and high resistance of trophectoderm cells against this drug were observed. The role of dihydrololate reductase at the preimplantation developmental stages, the resistance against aminopterin of the early rat embryos within the maternal organism and high sensitivity to this drug of the embryos in vitro are discussed. A conclusion is drawn on the presence of the barrier function of oviducts with respect to aminopterin.

Aminopterin

Generative model for the first cell fate bifurcation in mammalian development.

The first cell fate bifurcation in mammalian development directs cells toward either the trophectoderm (TE) or inner cell mass (ICM) compartments in pre-implantation embryos. This decision is regulated by the subcellular localization of a transcriptional co-activator YAP and takes place over several progressively asynchronous cleavage divisions. As a result of this asynchrony and variable arrangement of blastomeres, reconstructing the dynamics of the TE/ICM cell specification from fixed embryos is extremely challenging. To address this, we developed a live-imaging approach and applied it to measure pairwise dynamics of nuclear YAP and its direct target genes, CDX2 and SOX2, which are key transcription factors of the TE and ICM, respectively. Using these datasets, we constructed a generative model of the first cell fate bifurcation, which reveals the time-dependent statistics of the TE and ICM cell allocation. In addition to making testable predictions for the joint dynamics of the full YAP/CDX2/SOX2 motif, the model revealed the stochastic nature of the induction timing of the key cell fate determinants and identified the features of YAP dynamics that are necessary or sufficient for this induction. Notably, temporal heterogeneity was particularly prominent for SOX2 expression among ICM cells. As heterogeneities within the ICM have been linked to the initiation of the second cell fate decision in the embryo, understanding the origins of this variability is of key significance. The presented approach reveals the dynamics of the first cell fate choice and lays the groundwork for dissecting the next cell fate decisions in mouse development.

Animals

The survival and implantation of mouse blastocysts at varying degrees of reduced atmospheric pressure.

Pregnant mice were exposed to reduced atmospheric pressures ranging from 630 to 390 mm Hg during the pre-implantation and implantation periods and the numbers of embryos surviving 85 hours post coitum compared with those in litter-mate controls. Even at a pressure of 630 mm Hg (= 1,550 mm Hg) there was a significant fall in numbers of normal blastocysts and rise in abnormal forms before implantation, and implantation sites were reduced in number. The numbers of abnormal forms increased and implantation sites decreased at lower pressures, suggesting strongly that the hypoxia of reduce atmospheric pressure was responsible for the abnormalities observed. The pre-implantation period appears to be one during which the fertilised ovum is at particular risk, both of hypoxic damage and of failure to implant. Implantation may afford a degree of protection against hypoxia.

Animals

Mouse totipotent blastomere-like cells model embryogenesis from zygotic genome activation to post implantation.

Embryo development begins with zygotic genome activation (ZGA), eventually generating blastocysts for implantation. However, in vitro systems modeling the pre-implantation development are still absent and challenging. Here, we used mouse totipotent blastomere-like cells (TBLCs) to develop spontaneous differentiation and blastoid formation systems, respectively. We found Wnt signaling enabled the rapid expansion of TBLCs and the optimization of their culture medium. We successfully developed a TBLC-spontaneous differentiation system in which mouse TBLCs (mTBLCs) firstly converted into two types of ZGA-like cells (ZLCs) distinguished by Zscan4 expression. Surprisingly, Zscan4-, but not Zscan4+, ZLCs further passed through intermediate 4-cell and then 8-cell/morula stages to produce epiblast, primitive endoderm, and trophectoderm lineages. Significantly, single TBLCs underwent expansion, compaction, and polarization to efficiently generate blastocyst-like structures and even post-implantation egg-cylinder-like structures. Conclusively, we established TBLC-based differentiation and embryo-like structure formation systems to model early embryonic development, offering criteria for evaluating and understanding totipotency.

Animals

Localization of trophoblast-defined surface antigens during early mouse embryogenesis.

The binding pattern of a rabbit antiserum raised against mouse-ectoplacental-cone trophoblast on component cell populations in the pre-implantation and early post-implantation mouse embryo has been examined at the electron-microscope level using an immunoperoxidase-labelling technique. Binding was not detectable on the 1-cell stage, appeared at low levels at the 8-cell stage and was heavy on the trophectoderm and its trophoblast giant cell and extra-embryonic ectoderm descendants in the post-implantation embryo. In contrast, immunosurgically isolated 3 1/2-day inner cell masses (ICM) showed only slight labelling, whilst ICM derivatives in the 7 1/2-day embryo were unlabelled. The results indicate that the antiserum may be identifying a trophoblast-specific surface determinant(s), which appears with the differentiation of the trohectoderm and is maintained on some of the cell populations derived from this tissue at least until the early post-implantation stages.

Animals

The effect of exogenous gondadotropins on the pre-implantation period in the rat.

It is well established that the egg implantation in the rat is dependent on a surge of gonadotropin secretion on day 4 of pregnancy. The present experiments were designed in order to demonstrate the possible effect of exogenous administration of gonadotropins during pre-implantation period in pseudopregnant rats, made pregnant after egg transfer. Groups of pseudopregnant rats by sterile mating were injected with different doses of LH and FSH, between 9:00-12:00 A.M. on day 4 of pseudopregnancy; blastocysts were transferred to them in the morning of day 6, instead of day 5. The implantation rate and the number of young born at term were the criteria of the successful extension of the sensitive period of the endometrium by the exogenous gonadotropins. From the results obtained it can be concluded that LH alone acts on the uterine sensitivity by extension of the implantation period.

Animals