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

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.

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 101-121
Author(s):  
Hester P. M. Pratt

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.


Zygote ◽  
2000 ◽  
Vol 8 (3) ◽  
pp. 235-243 ◽  
Author(s):  
Pin-chi Tang ◽  
John D. West

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.


Development ◽  
1977 ◽  
Vol 40 (1) ◽  
pp. 83-89
Author(s):  
Lincoln V. Johnson ◽  
Patricia G. Calarco ◽  
Margaret L. Siebert

Alkaline phosphatase (AP) activity has been assayed in frozen sections of preimplantation mouse embryos by an azo-dye cytochemical method. The results indicate that during preimplantation mouse development AP activity is first expressed between the 8- and 16-cell stages and develops in all cells by the late morula stage. During blastocyst formation AP activity is lost or greatly reduced in trophoblast cells while activity is maintained in the inner cell mass.


Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 133-152
Author(s):  
Susan J. Kimber ◽  
M. Azim ◽  
H. Surani ◽  
Sheila C. Barton

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.


Development ◽  
1977 ◽  
Vol 40 (1) ◽  
pp. 143-157
Author(s):  
Paul S. Burgoyne ◽  
Thomas Ducibella

Mouse embryos (8-cell to early blastocyst) were denuded with pronase, and apposed in pairs which represented a wide range of stage combinations. These pairs either formed aggregates which differentiated into double-sized blastocysts, or they failed to aggregate. The 8–16-cell stages would not envelop late morulae/early blastocysts to form layered aggregates. This must mean that as the embryo differentiates into a blastocyst, the outer surface of the trophoblast loses its capacity for supporting cell spreading. The aggregation data also demonstrate that embryos almost completely lose their potential for aggregation at a very discrete stage in development – namely, between 8 and 9 h before blastocoel formation. It is argued that this is the stage at which the zonular tight junctional seal is completed, and that it is this physical barrier which prevents aggregation. It has been argued previously that the zonular tight junctional seal allows the creation of the special microenvironment which is necessary for the determination of the inner cells as inner cell mass. The completion of this seal 8–9 h before it is required for the formation of a blastocoel would provide a suitable time period for this cell determination to occur. The results obtained also relate to the technique of chimera production. Since the aim of this technique is to generate mice with mixed cell populations, it is important that the blastocyst formed following aggregation should have both cell lines present in the inner cell mass. This can best be assured by using relatively late morula stages (75 h post-HCG injection) since these will have already segregated their inner cells, but the incomplete seal will still allow aggregation to take place.


Development ◽  
1982 ◽  
Vol 67 (1) ◽  
pp. 51-58
Author(s):  
L. Izquierdo ◽  
M. I. Becker

Two-cell mouse embryos were cultured in vitro for different periods in a medium in which NaCl was partially replaced by LiCl at concentrations ranging from 1 to 30 mm. The relative cell number diminished according to increasing LiCl concentrations but the onset of blastulation was not affected, thus resulting in blastulae with fewer cells than normal and with a reduced or absent inner cell mass. Results are discussed in terms of the possible mechanisms involved and are related with the vegetalization induced by Li+ on early embryos of echinoderms and amphibia.


Development ◽  
1994 ◽  
Vol 120 (11) ◽  
pp. 3289-3299 ◽  
Author(s):  
C. Gueth-Hallonet ◽  
A. Santa-Maria ◽  
P. Verroust ◽  
B. Maro

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.


Development ◽  
1988 ◽  
Vol 104 (2) ◽  
pp. 219-234
Author(s):  
J.A. Emerson

The distribution of the cytokeratin network in the intact preimplantation mouse embryo and the role of cytokeratin filaments in trophectoderm differentiation were investigated by means of whole-mount indirect immunofluorescence microscopy and microinjection of anti-cytokeratin antibody. Assembled cytokeratin filaments were detected in some blastomeres as early as the compacted 8-cell stage. The incidence and organization of cytokeratin filaments increased during the morula stage, although individual blastomeres varied in their content of assembled filaments. At the blastocyst stage, each trophectoderm cell contained an intricate network of cytokeratin filaments, and examination of sectioned blastocysts confirmed that extensive arrays of cytokeratin filaments were restricted to cells of the trophectoderm. Microinjection of anticytokeratin antibody into individual mural trophectoderm cells of expanded blastocysts resulted in a dramatic rearrangement of the cytokeratin network in these cells. Moreover, antibody injection into 2-cell embryos inhibited assembly of the cytokeratin network during the next two days of development. Despite this disruption of cytokeratin assembly, the injected embryos compacted and developed into blastocysts with normal morphology and nuclear numbers. These results suggest that formation of an elaborate cytokeratin network in preimplantation mouse embryos is unnecessary for the initial stages of trophectoderm differentiation resulting in blastocyst formation.


Development ◽  
1986 ◽  
Vol 98 (1) ◽  
pp. 209-217
Author(s):  
G. F. Rands

The study describes an analysis of the development of mouse embryos halved at the 2-cell stage by the destruction of one blastomere, in comparison with control embryos of parallel derivation, at 2·5–13·5 days post coitum. The results showed that: (1) half embryos achieve size regulation some time between 7·5 and 10·5 days; (2) there is an indication that by 13·5 days half embryos may have again dropped back significantly in size relative to controls; (3) preregulation half embryos are slightly retarded developmental, but this does not wholly account for their smaller size: morphogenesis is not size-dependent; (4) early postimplantation half embryos contain a significantly decreased proportion of inner cell mass derivatives and increased proportion of trophectoderm derivatives when compared with controls. A comparison is also made between the up-regulation of half embryos and the down-regulation of aggregate embryos, and it is suggested that size regulation may occur by delaying a change in the normal growth rate.


Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 241-261
Author(s):  
M. H. Johnson ◽  
J. Chakraborty ◽  
A. H. Handyside ◽  
K. Willison ◽  
P. Stern

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.


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