The effect of prolonged decompaction on the development of the preimplantation mouse embryo

Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 241-261
Author(s):  
M. H. Johnson ◽  
J. Chakraborty ◽  
A. H. Handyside ◽  
K. Willison ◽  
P. Stern
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Rabbit Antiserum ◽  
Embryonal Carcinoma Cell ◽  
Cell Stage ◽  
Embryonal Carcinoma Cell Line ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Cell Embryo ◽  
Relationship Of

A rabbit antiserum to a mouse embryonal carcinoma cell line blocks compaction of cleaving mouseembryos. Cell division is not affected up to the 32-cell stage but intracellular junctions fail to develop. Removal of the antibody at this stage permits compaction to occur and a normal blastocyst develops. Prolonged decompaction beyond the 32-cell embryo results in an increasing proportion of malformed blastocysts in which trophectodermal cells predominate and functional inner cell mass (ICM) cells are reduced or absent. The relationship of compaction to the generation of ICM and trophectoderm lineages in the intact embryo is discussed.

Interactions of blastomeres suggest changes in cell surface adhesiveness during the formation of inner cell mass and trophectoderm in the preimplantation mouse embryo

Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 133-152
Author(s):  
Susan J. Kimber ◽  
M. Azim ◽  
H. Surani ◽  
Sheila C. Barton
Keyword(s):  
Inner Cell Mass ◽  
Single Cells ◽  
Cell Mass ◽  
Cell Stage ◽  
Trophectoderm Cells ◽  
Adhesive Surface ◽  
Preimplantation Mouse Embryo ◽  
Divergent Differentiation ◽  
Preimplantation Mouse ◽  
The Difference

Whole 8-cell morulae can be aggregated with isolated inner cell masses from blastocysts. On examining semithin light microscope sections of such aggregates we found that cells of the morula changed shape and spread over the surface of the ICM, thus translocating it to the inside of the aggregate. Using single cells from 8-cell embryos in combination with single cells from other stage embryos or isolated ICMs we show that 1/8 blastomeres spread over other cells providing a suitably adhesive surface. The incidence of spreading is high with inner cells from 16-cell embryos (56 %) and 32-cell embryos (62%) and isolated inner cell masses (64%). In contrast, the incidence of spreading of 1/8 blastomeres is low over outer cells from 16-cell embryos (26%) and 32-cell embryos (13%). Blastomeres from 8-cell embryos do not spread over unfertilized 1-cell eggs, 1/2 or 1/4 cells or trophectoderm cells contaminating isolated ICMs. When 1/8 cells are aggregated in pairs they flatten on one another (equal spreading) as occurs at compaction in whole 8-cell embryos. However, if 1/8 is allowed to divide to 2/16 in culture one of the cells engulfs the other (51-62/ pairs). Based on the ideas of Holtfreter (1943) and Steinberg (1964,1978) these results are interpreted to indicate an increase in adhesiveness at the 8-cell stage as well as cytoskeletal mobilization. Following the 8-cell stage there is an increase in adhesiveness of inside cells while the outside cells decrease in adhesiveness. The difference in adhesiveness between inside and outside cells in late morulae is probably central to the divergent differentiation of (inner) ICM and (outer) trophectoderm cell populations.

Membrane organization in the preimplantation mouse embryo

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 101-121
Author(s):  
Hester P. M. Pratt
Keyword(s):  
Mouse Embryo ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Freeze Fracture ◽  
Molecular Organization ◽  
Surface Polarity ◽  
Cell Stage ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Polarized Epithelium

The preimplantation mouse blastocyst consists of two differentiated tissues, the trophectoderm (a structurally and functionally polarized epithelium) and the inner cell mass. The divergence of these two cell types can be traced back to a contact dependent polarization of the surface and cytoplasm at the 8-cell stage. Membrane/cytocortical organization during this preimplantation period has been studied using freeze fracture in conjunction with the sterol-binding antibiotic filipin in an attempt to discern the molecular basis and origin of these surface asymmetries. The distribution of filipin reactivity within the different membrane domains showed that the surface polarity exhibited by trophectoderm and by blastomeres of the 8-cell stage is underlain by a heterogeneity in molecular organization of the membrane/cytocortex which may originate prior to the appearance of any overt surface polarity. The results are discussed in terms of the likely basis of this membrane/cytocortical asymmetry, its probable origins and the use of the preimplantation mouse embryo as a model system for studying the assembly of a polarized epithelium.

The effects of embryo stage and cell number on the composition of mouse aggregation chimaeras

Zygote ◽  
2000 ◽  
Vol 8 (3) ◽  
pp. 235-243 ◽  
Author(s):  
Pin-chi Tang ◽  
John D. West
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Number ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Advanced Embryo ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Cell Embryo ◽  
Embryo Stage

Studies with intact preimplantation mouse embryos and some types of chimaeric aggregates have shown that the most advanced cells are preferentially allocated to the inner cell mass (ICM) rather than the trophectoderm. Thus, differences between 4-cell and 8-cell stage embryos could contribute to the tendency for tetraploid cells to colonise the trophectoderm more readily than the ICM in 4-cell tetraploid[harr ]8 cell diploid chimaeras. The aim of the present study was to test whether 4-cell stage embryos in 4-cell diploid[harr ]8-cell diploid aggregates contributed equally to all lineages present in the E12.5 conceptus. These chimaeras were compared with those produced from standard aggregates of two whole 8-cell embryos and aggregates of half an 8-cell embryo with a whole 8-cell embryo. As expected, the overall contribution of 4-cell embryos was lower than that of 8-cell embryos and similar to that of half 8-cell stage embryos. In the 4-cell[harr ]8-cell chimaeras the 4-cell stage embryos did not contribute more to the trophectoderm than the ICM derivatives. Thus, differences between 4-cell and 8-cell embryos cannot explain the restricted tissue distribution of tetraploid cells previously reported for 4-cell tetraploid[harr ]8-cell diploid chimaeras. It is suggested that cells from the more advanced embryo are more likely to contribute to the ICM but, for technical reasons, are prevented from doing so in simple aggregates of equal numbers of whole 4-cell and whole 8-cell stage embryos.

scholarly journals Changes in the distribution of a spectrin-like protein during development of the preimplantation mouse embryo.

1985 ◽  
Vol 100 (1) ◽  
pp. 333-336 ◽  
Author(s):  
J S Sobel ◽  
M A Alliegro
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Contact ◽  
Apical Region ◽  
Cell Stage ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Alpha Spectrin ◽  
Avian Erythrocyte ◽  
Cell Cell

The mouse blastocyst expresses a 240,000-mol-wt polypeptide that cross-reacts with antibody to avian erythrocyte alpha-spectrin. Immunofluorescence localization showed striking changes in the distribution of the putative embryonic spectrin during preimplantation and early postimplantation development. There was no detectable spectrin in either the unfertilized or fertilized egg. The first positive reaction was observed in the early 2-cell stage when a bright band of fluorescence delimited the region of cell-cell contact. The blastomeres subsequently developed continuous cortical layers of spectrin and this distribution was maintained throughout the cleavage stages. A significant reduction in fluorescence intensity occurred before implantation in the apical region of the mural trophoblast and the trophoblast outgrowths developed linear arrays of spectrin spots that were oriented in the direction of spreading. In contrast to the alterations that take place in the periphery of the embryo, spectrin was consistently present in the cortical cytoplasm underlying regions of contact between the blastomeres and between cells of the inner cell mass. The results suggest a possible role for spectrin in cell-cell interactions during early development.

The nature and distribution of serologically detectable alloantigens on the preimplantation mouse embryo

Development ◽  
1976 ◽  
Vol 35 (1) ◽  
pp. 59-72
Author(s):  
Audrey L. Muggleton-Harris ◽  
Martin H. Johnson
Keyword(s):  
Mouse Embryo ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Convincing Evidence ◽  
Mouse Embryos ◽  
Cell Type ◽  
Cell Stage ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse

The nature and distribution of surface alloantigens on preimplantation mouse embryos has been examined by immunofluorescence. Non-H-2 alloantigens were detected at allstages examined, from the 2-cell to the 4½-day blastocyst. Cleaving blastomeres, inner cell mass cells and cells of the primary trophectoderm were all positive. In F1 embryos maternalnon-H-2 alloantigens were detectable at all stages, whereas paternal antigens first became evident at the 6- to 8-cell stage. No convincing evidence of the presence of alloantigens associated with the H-2 haplotype was found at any stage or on any cell type, suggesting that if these antigens are present they are in low quantity or are masked.

When and how does cell division order influence cell allocation to the inner cell mass of the mouse blastocyst?

Development ◽  
1987 ◽  
Vol 100 (2) ◽  
pp. 325-332
Author(s):  
C.L. Garbutt ◽  
M.H. Johnson ◽  
M.A. George
Keyword(s):  
Cell Division ◽  
Cell Population ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
The Other ◽  
Mouse Blastocyst ◽  
Short Term ◽  
Cell Stage ◽  
Cell Embryo

Aggregate 8-cell embryos were constructed from four 2/8 pairs of blastomeres, one of which was marked with a short-term cell lineage marker and was also either 4 h older (derived from an early-dividing 4-cell) or 4 h younger (derived from a late-dividing 4-cell) than the other three pairs. The aggregate embryos were cultured to the 16-cell stage, at which time a second marker was used to label the outside cell population. The embryos were then disaggregated and each cell was examined to determine its labelling pattern. From this analysis, we calculated the relative contributions to the inside cell population of the 16-cell embryo of older and younger cells. Older cells were found to contribute preferentially. However, if the construction of the aggregate 8-cell embryo was delayed until each of the contributing 2/8 cell pairs had undergone intercellular flattening and then had been exposed to medium low in calcium to reverse this flattening immediately prior to aggregation, the advantage possessed by the older cells was lost. These results support the suggestion that older cells derived from early-dividing 4-cell blastomeres contribute preferentially to the inner cell mass as a result of being early-flattening cells.

Cosegregation of monoclonal antibody reactivity and cell behaviour in the mouse preimplantation embryo

Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 261-278
Author(s):  
Beverley J. Randle
Keyword(s):  
Monoclonal Antibody ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Antigen Expression ◽  
Cell Stage ◽  
Cell Behaviour ◽  
Minor Population ◽  
Cell Embryo ◽  
Surface Labelling

Expression of an antigen, recognized by a monoclonal antibody raised against PCI 3 embryonal carcinoma, is described in mouse preimplantation embryogenesis. The antigen is found in the cytoplasm of ovulated ova and is first noted on the cell surface of the 1-cell embryo 20 h post-ovulation. Surface labelling of blastomeres is uniform until the 8-cell stage when antigen expression becomes polarized along the radial axis of the embryo. Two major populations of blastomeres are distinguishable on division to the 16-cell morula. Dissociation of morulae in calcium-free medium yields large, polar, antigen-positive cells and small apolar cells with reduced levels of detectable antigen. A third, minor population of small, antigen-negative cells is also found in vivo. Large and small blastomeres differ in their ability to relocate within the embryo when aggregated with intact 16-cell-stage embryos. The small blastomeres of the 16-cell morula contribute significantly to the inner cell mass while the large antigen-positive cells are found only in the trophectoderm.

Features of cell lineage in preimplantation mouse development

Development ◽  
1978 ◽  
Vol 48 (1) ◽  
pp. 53-72
Author(s):  
C. F. Graham ◽  
Z. A. Deussen
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Lineage ◽  
The Other ◽  
Mouse Development ◽  
Cell Stage ◽  
Daughter Cells ◽  
Preimplantation Mouse ◽  
Peripheral Cells

The cell lineage of the mouse was studied from the 2-cell stage to the blastocyst. Lineage to the 8-cell stage was followed under the microscope. Each cell from the 2-cell stage divided to form two daughter cells which remained attached. Subsequently, these two daughters each produced two descendants; one of these descendants regularly lay deep in the structure of the embryo while the other was peripheral. Lineage to the blastocyst was followed by injecting oil drops into cells at the 8-cell stage, and then following the segregation of these drops into the inner cell mass and trophectoderm. Between the 8-cell stage and the blastocyst, the deep cells contributed more frequently to the inner cell mass than did the peripheral cells.

scholarly journals Embryo cell allocation patterns are not altered by biopsy but can be linked with further development

Reproduction ◽  
2017 ◽  
Vol 154 (6) ◽  
pp. 807-814
Author(s):  
L P Sepulveda-Rincon ◽  
N Islam ◽  
P Marsters ◽  
B K Campbell ◽  
N Beaujean ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Mouse Strains ◽  
Random Pattern ◽  
Cell Stage ◽  
Cell Counts ◽  
Cell Embryo ◽  
Allocation Patterns ◽  
Further Development

It has been suggested that first embryo cleavage can be related with the embryonic–abembryonic axis at blastocyst stage in mice. Thus, cells of the 2-cell embryo might be already biased to form the inner cell mass or trophectoderm. This study was conducted to observe the possible effects of embryo biopsy on cell allocation patterns during embryo preimplantation in two different mouse strains and the effects of these patterns on further development. First, one blastomere of the 2-cell embryo was injected with a lipophilic tracer and cell allocation patterns were observed at blastocyst stage. Blastocysts were classified into orthogonal, deviant or random pattern. For the first experiment, embryos were biopsied at 8-cell stage and total cell counts (TCC) were annotated. Furthermore, non-biopsied blastocysts were transferred into foster mothers. Then, pups and their organs were weighed two weeks after birth. Random pattern was significantly recurrent (≈60%), against orthogonal (<22%) and deviant (<22%) patterns among groups. These patterns were not affected by biopsy procedure. However, TCC on deviant embryos were reduced after biopsy. Moreover, no differences were found between patterns for implantation rates, litter size, live offspring and organ weights (lungs, liver, pancreas and spleen). However, deviant pups presented heavier hearts and orthogonal pups presented lighter kidneys among the group. In conclusion, these results suggest that single blastomere removal does not disturb cell allocation patterns during pre-implantation. Nonetheless, the results suggest that embryos following different cell allocation patterns present different coping mechanisms against in vitro manipulations and further development might be altered.

Gp330 is specifically expressed in outer cells during epithelial differentiation in the preimplantation mouse embryo

Development ◽  
1994 ◽  
Vol 120 (11) ◽  
pp. 3289-3299 ◽  
Author(s):  
C. Gueth-Hallonet ◽  
A. Santa-Maria ◽  
P. Verroust ◽  
B. Maro
Keyword(s):  
Mouse Embryo ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Transcriptional Level ◽  
Cell Stage ◽  
Preimplantation Mouse ◽  
Extensive Cell ◽  
High Level ◽  
Endocytic Activity

During preimplantation development of the mouse embryo, a layer of outer cells differentiates into a perfect epithelium, the trophectoderm. The divergence between the trophectoderm and the inner cell mass takes place from the 8-cell stage to the 64-cell stage and precedes their commitment at the blastocyst stage. In this work, we have investigated the expression of gp330, a 330 × 10(3) M(r) glycoprotein found in clathrin-coated areas of the plasma membrane of some epithelial cells characterized by a high level of endocytic activity. Our results show that gp330 is first synthesized in 16-cell stage embryos and that its appearance is restricted to outer cells until the blastocyst stage. Furthermore, its expression is repressed in inner cells at a post-transcriptional level, probably through the development of extensive cell-cell contacts.

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