The Influence of the Uterine Environment on the Development of Mouse Eggs

Development ◽  
1962 ◽  
Vol 10 (4) ◽  
pp. 496-506
Author(s):  
D. R. S. Kirby
Keyword(s):  
Anterior Chamber ◽  
Abdominal Cavity ◽  
Cell Stage ◽  
Kidney Capsule ◽  
Uterine Environment ◽  
Series Of Experiments ◽  
Mouse Eggs ◽  
Uterine Development

From the literature and the results of some preliminary experiments by the author (Kirby, 1960), it appeared that the ability of mouse eggs to develop into embryos in an extra-uterine site was at least partially dependent on the stage of the egg when transplanted. In most previous studies on the extra-uterine development of mouse eggs only tubal eggs have been transplanted. Fawcett, Wislocki, & Waldo (1947) transferred 8–10-cell-stage eggs to the anterior chamber of the eye, 3 or 4 eggs being injected in each case. In another series of experiments reported in the same paper the oviducts were transected, thereby causing the descending eggs to pass into the abdominal cavity. Fawcett (1950) transplanted 2–10-cell-stage eggs to a position beneath the kidney capsule, and, to compensate for possible loss, several eggs were usually introduced. Jones 1951) transplanted 2–10-cell-stage eggs to the eye and beneath the kidney capsule.

Extra-uterine development of mouse blastocysts cultured in vitro from early cleavage stages

Development ◽  
1968 ◽  
Vol 20 (3) ◽  
pp. 391-400
Author(s):  
W. D. Billington ◽  
C. F. Graham ◽  
Anne McLaren
Keyword(s):  
Blastocyst Stage ◽  
Embryonic Tissue ◽  
Early Cleavage ◽  
Embryonic Tissues ◽  
Complete Development ◽  
Uterine Environment ◽  
Cleavage Stages ◽  
Mouse Eggs ◽  
Uterine Development

Many experiments indicate that oviducal mouse eggs are unable to form embryonic tissues following transplantation to extra-uterine sites; they give rise only to trophoblast and extra-embryonic membranes (Fawcett, Wislocki & Waldo, 1947; Fawcett, 1950; Chester-Jones, 1951; Whitten, 1958; Kirby, 1962; Billington, 1965). Possible rare exceptions exist in the reports by Runner (1947) and Stevens (1967). In contrast, about a quarter of uterine blastocysts form morphologically normal embryos when similarly transplanted (Kirby, 1963; Billington, 1965). The conditions under which the mouse eggs reach the blastocyst stage may also have developmental consequences. Eggs cultured in vitro to the blastocyst stage (Whitten, 1956) or tube-locked blastocysts (Kirby, 1962) did not develop embryonic tissue in extra-uterine sites, but were capable of complete development following transfer to a receptive uterus. These findings suggest that mouse eggs must be subjected to the uterine environment if they are to ‘realize totipotency’ in an extra-uterine site (Kirby, 1962, 1965).

Intercellular communication in the early embryo of the ascidian Ciona intestinalis

Development ◽  
1988 ◽  
Vol 102 (1) ◽  
pp. 55-63 ◽  
Author(s):  
F. Serras ◽  
C. Baud ◽  
M. Moreau ◽  
P. Guerrier ◽  
J.A.M. Van den Biggelaar
Keyword(s):  
Intercellular Communication ◽  
Lucifer Yellow ◽  
Ciona Intestinalis ◽  
Early Embryo ◽  
Cell Stage ◽  
Early Embryos ◽  
Junctional Conductance ◽  
Voltage Dependent ◽  
Cytoplasmic Bridges ◽  
Series Of Experiments

We have studied the intercellular communication pathways in early embryos of the ascidian Ciona intestinalis. In two different series of experiments, we injected iontophoretically the dyes Lucifer Yellow and Fluorescein Complexon, and we analysed the spread of fluorescence to the neighbouring cells. We found that before the 32-cell stage no dye spread occurs between nonsister cells, whereas sister cells are dye-coupled, possibly via cytoplasmic bridges. After the 32-cell stage, dye spread occurs throughout the embryo. However, electrophysiological experiments showed that nonsister cells are ionically coupled before the 32-cell stage. We also found that at the 4-cell stage junctional conductance between nonsister cells is voltage dependent, which suggests that conductance is mediated by gap junctions in a way similar to that observed in other embryos.

240 DIFFERENCES IN GLOBAL TRANSCRIPTOME PROFILE OF BOVINE BLASTOCYSTS DERIVED FROM SUPEROVULATED OR SYNCHRONIZED CYCLIC HEIFERS

2010 ◽  
Vol 22 (1) ◽  
pp. 278
Author(s):  
A. Gad ◽  
M. Hoelker ◽  
F. Rings ◽  
N. Ghanem ◽  
D. Salilew-Wondim ◽  
...  
Keyword(s):  
Data Analysis ◽  
Transcript Abundance ◽  
R Package ◽  
Transcriptome Profile ◽  
Metabolic Processes ◽  
Cell Stage ◽  
Endoscopic Method ◽  
Response To Stress ◽  
Uterine Environment ◽  
Transfer Method

Estrus synchronization and superovulation are the most widely used procedures in embryo transfer technology. However, changes in the oviduct and uterine environment due to these procedures and the subsequent influence on embryos have not yet been investigated. This study was con- ducted to investigate the effect of oviduct environment of only synchronized or superovulated cyclic heifers on the gene expression profile of blastocysts. Bovine Affymetrix array analysis was performed using 2 groups of blastocysts. The first group was bovine blastocysts produced after superovulation of Simmental heifers (n = 9) using 8 consecutive FSH injections over 4 days in decreasing doses (in total, 300-400 mg of FSH equivalent according to body weight) and flushed at Day 7 by nonsurgical endoscopic method. The second group was bovine blastocysts derived from synchronized Simmental heifers (n = 4) after transfer of 2-cell stage embryos from superovulated donor Simmental heifers (n = 9) by nonsurgical transvaginal endoscopy tubal transfer method. Total RNA was extracted from 3 pools of embryos from each experimental group (6 embryos per pool). A total of 6 biotin-labeled cRNA samples were hybridized on 6 bovine Affymetrix arrays. Data analysis was performed using LIMMA written on R package, which maintained the Bioconductor. Array data analysis revealed a total of 454 transcripts to be differen- tially expressed (P < 0.05, fold change >2) between the 2 groups. Of these, 429 and 25 were up- and down-regulated, respectively, in blastocysts derived from superovulated heifers compared with those derived from synchronized animals. Genes involved in response to stress (HSPA14 and HSPE1), cellular and metabolic processes (CPSF3, ATPIF1, POMP, and MDH2), translation (RPS17, EEF1B2, and EIF4E), and cell commu- nication (FN1, KRT18, and DSG2) were found to be enriched in blastocysts derived from superovulated animals. On the other hand, protein metabolic processes related genes (CLGN) were found to be enriched in blastocysts derived from the synchronized group. The KEGG analysis of the differentially expressed genes showed that the ribosome and oxidative phosphorylation pathways are the dominant pathways and genes involved in these pathways are greatly abundant in the blastocysts derived from superovulated animals. Quantitative real-time PCR has confirmed the transcript abundance of 7 out of 8 genes selected for validation. In conclusion, blastocysts cultured in synchronized animals post 2-cell stage showed significant differences in transcriptome profile compared with their counterparts that remained in superovulated heifers until Day 7. Further functional analysis of some selected candidate genes could give new insights into mechanisms regulating the ability of embryos to survive after transfer.

Further Observations on Experimental Toxi-Infection of the Central Nervous System

1918 ◽  
Vol 64 (264) ◽  
pp. 18-29
Author(s):  
David Orr ◽  
Rows
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Experimental Work ◽  
Abdominal Cavity ◽  
The Central Nervous System ◽  
Series Of Experiments

This communication is a continuation of our experimental work on the action of bacterial poisons upon the nervous system.In 1914 (2), after several series of experiments, we drew attention to the differences between lymphogenous and haematogenous infection. The first was induced by infecting the ascending lymph stream of nerves ; the second by placing celloidin capsules containing a culture of bacteria in the abdominal cavity.

98 EXPERIMENTAL TRANSFER OF BOVINE IVF-DERIVED 32-CELL STAGE EMBRYOS INTO THE UTERUS: ENVIRONMENTAL EFFECTS ON DEVELOPMENTAL CHARACTERISTICS

2016 ◽  
Vol 28 (2) ◽  
pp. 179
Author(s):  
M. Hoelker ◽  
D. Salilew-Wondim ◽  
F. Rings ◽  
D. Tesfaye ◽  
K. Schellander
Keyword(s):  
Culture Media ◽  
Uterine Cavity ◽  
Blastocyst Stage ◽  
Considerable Proportion ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Developmental Rates ◽  
Uterine Environment

Usually, in vitro-produced bovine embryos are cultured in vitro in static culture systems for 7 to 9 days in media composed according the oviducal fluid although it is well accepted that around Day 4.5–5 the bovine embryo enters the uterine cavity, providing environmental conditions different from the oviduct. Therefore, one has to raise the question whether changing culture media properties after Day 5 of culture could have beneficial effects on early development of bovine embryos. To answer that question, we transferred bovine IVF derived 32-cell stage embryos into the uterine cavity of synchronized recipients. All embryos had been matured and fertilized under routine standard conditions and were cultured in synthetic oviducal fluid supplemented with essential and nonessential amino acids (SOFaa) supplemented with either 0.3% fatty acid free bovine serum albumin (BSAfaf/Uterus) or 10% serum (serum/uterus) at 38.5°C, 5% O2, and 5% CO2 in humidified air prior transfer into the uterine environment, allowing further development to the blastocyst stage within the physiological environment prior recollection at Day 7 by routine uterine flushing followed by comparison with statically in vitro-developed embryos cultured in media supplemented with serum (serum/serum group) or BSAfaf (BSAfaf/BSAfaf group). All in all, a total of 1031 in vitro-derived 32-cell stage embryos were transferred to 21 synchronized Simmental recipient heifers. Of these, a total of 680 embryos (66%) could be recollected at Day 7. Embryos of the serum/serum group reached a higher blastocyst rate compared with embryos of the BSAfaf/BSAfaf group (68% v. 41%; P < 0.05, ANOVA, Tukey test), whereas the developmental rate to the blastocyst stage did not differ after 9 days of in vitro culture, indicating higher developmental kinetics of bovine 32-cell stage embryos when culture media is supplemented with serum. Moreover, embryos of the serum/uterus group reached significantly lower developmental rates to the blastocyst stage until Day 7 compared with embryos of the serum/serum group (12.9% v. 68.4%). Likewise, embryos in the BSAfaf/uterus group reached significantly lower developmental rates to the blastocyst stage until Day 7 compared with embryos in the BSAfaf/BSAfaf group (16.0% v. 40.1%). When allowed to develop for additional 48h in vitro, developmental rates to the blastocyst stage at Day 9 were still higher in BSAfaf/BSAfaf treatment compared with the BSAfaf/uterus treatment (91.4% v. 74.4%) and the serum/serum treatment compared with the serum/uterus treatment (92.5% v. 56.0%). Taken together, the results of our study demonstrate that uterine transfer of bovine 32-cell stage embryos results in reduction of developmental kinetics as well as lower developmental rates compared with embryos statically cultured in vitro. That might indicate, that a considerable proportion of bovine 32-cell stage embryos might not be able to adapt to the uterine environment.

Timing of the First Cleavage Division of Haploid Mouse Eggs, and the Duration of its Component Stages

1973 ◽  
Vol 13 (2) ◽  
pp. 553-566 ◽  
Author(s):  
M. H. KAUFMAN
Keyword(s):  
Polar Body ◽  
Time Lapse ◽  
Follicle Cells ◽  
Cleavage Division ◽  
Cell Stage ◽  
Cell Embryo ◽  
Second Polar Body ◽  
Polar Body Extrusion ◽  
Mouse Eggs

Mouse eggs were activated by treatment with hyaluronidase which removed the follicle cells, followed by culture in vitro, and examined at the first cleavage mitosis. Second polar body extrusion usually occurred and haploid parthenogenesis was initiated. Air-dried chromosome preparations were made between 11 and 15.5 h after activation. Out of the 308 eggs examined 74 had already progressed to the 2-cell stage; the remaining 234 at the 1-cell stage were examined in detail. All chromosome preparations of the first cleavage mitosis were classified into groups corresponding with the stages of prometaphase, metaphase (early or ‘pre-chromatid’, ‘chromatid’ and ‘late chromatid’) and anaphase. An indirect estimate was made of the duration of the first cleavage mitosis and of its component stages from the incidence of stages observed at different time intervals after activation. Similar eggs were also observed at 37 °C by time-lapse cine-photography and the interval between the disappearance of the pronucleus to the beginning of telophase of the first cleavage division was determined. The results of timing studies on the haploid eggs were compared with results obtained from similar observations on the first cleavage division of fertilized eggs which would of course normally be diploid. Artificially activated eggs with 2 pronuclei, resulting from second polar body suppression, were also examined, and serial chromosome preparations during mitosis showed that the 2 pronuclear chromosome groups unite on the first cleavage spindle and divide to give a hetero-zygous diploid 2-cell embryo.

Timing of the First Cleavage Division of the Mouse and the Duration of its Component Stages: A Study of Living and Fixed Eggs

1973 ◽  
Vol 12 (3) ◽  
pp. 799-808
Author(s):  
M. H. KAUFMAN
Keyword(s):  
Close Correlation ◽  
Time Lapse ◽  
Culture Conditions ◽  
Effect Of Temperature ◽  
Time Intervals ◽  
Cleavage Division ◽  
Cell Stage ◽  
Mitotic Time ◽  
Mouse Eggs

Fertilized mouse eggs were examined between 27 and 34 h after the superovulating injection of human chorionic gonadotrophin (HCG). Out of 1334 eggs examined, 432 were already at the 2-cell stage; the remaining 902 at the 1-cell stage were examined in detail. All chromosome preparations of the first cleavage mitosis were classified into groups corresponding with the stages of prometaphase (early and late), metaphase (early or ‘prechromatid’, ‘chromatid’ and ‘late chromatid’) and anaphase. An indirect estimate was made of the duration of the first cleavage mitosis and of its component stages from the incidence of stages observed at different time intervals after the HCG injection. Fertilized eggs were also observed at 37°C by time-lapse cine-photography and the interval between the disappearance of the pronuclei and the beginning of telophase of the first cleavage division was determined. The progress of eggs fertilized in vitro was also observed under normal culture conditions. A close correlation was observed between the indirect method of assessing the mitotic time and the direct values obtained from the studies on time-lapse and in vitro culture. The effect of temperature on the mitotic time was also examined by the time-lapse method.

The development of teratomas from intratesticular grafts of tubal mouse eggs

Development ◽  
1968 ◽  
Vol 20 (3) ◽  
pp. 329-341
Author(s):  
Leroy C. Stevens
Keyword(s):  
Anterior Chamber ◽  
Inbred Mice ◽  
Germ Layers ◽  
Embryonic Tissues ◽  
Embryonic Shield ◽  
Mouse Eggs

Grafts of cleaving tubal ova from non-inbred mice to ectopic sites usually result in growths composed of extra-embryonic but not embryonic tissues (Fawcett, Wisloki & Waldo, 1947; Fawcett, 1950; Jones, 1951; Whitten 1958; Kirby, 1960, 1962a; Billington, 1965; and others). Runner (1947) grafted tubal mouse ova to the anterior chamber of the eye and one developed the three primary germ layers and then regressed, probably because the host and donor were histo-incompatible. This is the only report of an ectopically grafted pre-uterine egg that developed intra-embryonic derivatives. Kirby (1962b, 1965) grafted oviducal segmenting mouse eggs to the kidney and obtained only trophoblast and extra-embryonic membranes. He concluded that a ‘uterine factor’ is necessary for the development of intra-embryonic structures from mouse eggs. Kirby (1965) and Billington (1965) grafted morulae and blastocysts to the testis, and the morulae never gave rise to embryonic shield derivatives.

Reduced gap junctional communication is associated with the lethal condition characteristic of DDK mouse eggs fertilized by foreign sperm

Development ◽  
1987 ◽  
Vol 101 (3) ◽  
pp. 449-459 ◽  
Author(s):  
M. Buehr ◽  
S. Lee ◽  
A. McLaren ◽  
A. Warner
Keyword(s):  
Gap Junctions ◽  
Lucifer Yellow ◽  
Blastocyst Stage ◽  
Weak Base ◽  
Cell Stage ◽  
Gap Junctional Communication ◽  
Junctional Communication ◽  
Condition Characteristic ◽  
Gap Junctional ◽  
Mouse Eggs

Communication through gap junctions was examined in 8-cell zygotes generated by fertilization of eggs of the DDK inbred strain of mice with spermatozoa of the C3H strain. These zygotes spontaneously begin to extrude cells at the late 16-cell stage and 95% die by the blastocyst stage. The transfer of Lucifer Yellow between cells of DDK/C3H zygotes that had not yet begun to express the defect was significantly slower than in DDK/DDK controls or in controls from other strains. Treatment with the weak base methylamine, to raise intracellular pH, speeded the transfer of Lucifer in all strains; transfer between cells of DDK/C3H zygotes became as fast as that between cells of control zygotes. DDK/C3H zygotes cultured in methylamine either from the 4- to 8-cell stage to the early 16-cell stage (19h) or from the early to the late 16-cell stage (6 h) showed significant rescue to the blastocyst stage. Once spontaneous decompaction of cells from DDK/C3H zygotes had begun (the late 16-cell stage onwards) methylamine treatment was no longer able to bring about rescue. We conclude that zygotes developed from eggs of the DDK strain fertilized by foreign spermatozoa are characterized physiologically by defective gap junctional communication. Improving gap junctional communication is sufficient to allow many zygotes to maintain the compacted state, suggesting a link between compaction and communication through gap junctions.

The effect of the nucleocytoplasmic ratio on protein synthesis and expression of a stage-specific antigen in early cleaving mouse embryos

Development ◽  
1987 ◽  
Vol 99 (4) ◽  
pp. 481-491 ◽  
Author(s):  
U. Petzoldt ◽  
A. Muggleton-Harris
Keyword(s):  
Protein Synthesis ◽  
Specific Antigen ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Specific Gene ◽  
Cell Stage ◽  
Two Dimensional Gel Electrophoresis ◽  
Morula Stage ◽  
Early Mouse ◽  
Mouse Eggs

The nucleocytoplasmic ratio of fertilized mouse eggs was manipulated by removing or injecting cytoplasm by micropipette, and bisection of denuded eggs to obtain both pronuclei in one half of the eggs cytoplasm. The experimental eggs were capable of cleavage to the morula stage and, in some instances, developed to the blastocyst stage similar to unmanipulated eggs. The removal of large quantities of cytoplasm by micropipette and injecting them into a recipient egg did not provide sufficient numbers of viable eggs, whereas transfer of smaller quantities (about a quarter of the cytoplasm) was less deleterious, at least for recipient eggs. However, the alteration of the nucleocytoplasmic ratio by this method was not of the correct magnitude for the purpose of this experiment. Therefore, bisection was the preferred method whereby the nucleocytoplasmic ratio was doubled. This resulted in both pronuclei residing in one half of the egg's cytoplasm. Half eggs with one pronucleus (haploid) but retaining a nucleocytoplasmic ratio similar to unmanipulated control eggs served as additional controls for the bisection experiments. Protein synthesis was analysed by two-dimensional gel electrophoresis, showing that the 2-cell- and 4-cell-stage bisected embryos with double and normal nucleocytoplasmic ratio expressed equivalent protein synthesis patterns as control embryos of the same stage. Likewise, the stage-specific surface antigen SSEA-1 did not appear before the 6- to 8-cell stage. Also in cytoplasm transfer experiments, there was no indication that altering the nucleocytoplasmic ratio in either direction changed the timing of stage-specific gene expression. These results support the idea that stage-specific gene activity during early mouse cleavage might proceed in parallel to DNA replication cycles and is independent of the nucleocytoplasmic ratio.

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