Sperm penetration in parthenogenetic mouse embryos triggers a plasma membrane block to polyspermy

Zygote ◽  
1993 ◽  
Vol 1 (3) ◽  
pp. 237-242 ◽  
Author(s):  
Marek Maleszewski ◽  
Anna Bielak
Keyword(s):  
Plasma Membrane ◽  
Cell Membrane ◽  
Sperm Cell ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Cell Cycles ◽  
Sperm Penetration ◽  
Membrane Components ◽  
Parthenogenetic Mouse Embryos ◽  
Parthenogenetic Embryos

SummaryMouse oocytes activated parthenogenetically do not generate a plasma membrane block against spermatozoa over the first three cell cycles. We show that they lose this fusibility spontaneously at the 8-cell stage. Insemination of 1-cell parthenogenetic embryos induces loss of fusibility earlier, at the 2-cell stage. This observation suggests that incorporation of the sperm cell membrane components into the oolemma may be responsible for the development of the membrane block.

Complete preimplantation development in culture of parthenogenetic mouse embryos

Development ◽  
1976 ◽  
Vol 35 (1) ◽  
pp. 179-190
Author(s):  
Matthew H. Kaufman ◽  
Leo Sachs
Keyword(s):  
Polar Body ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Haploid Nucleus ◽  
Second Polar Body ◽  
Haploid Embryos ◽  
Parthenogenetic Mouse Embryos ◽  
Parthenogenetic Embryos ◽  
Better Than

The present experiments were undertaken to determine whether, in parthenogenesis, heterozygous embryos develop better than homozygous embryos. Such experiments may provide an approach to elucidating whether fertilized embryos develop better than parthenogenetic ones because of heterozygosity, or if the sperm provides another contribution necessary for complete embryonic development. The parthenogenetic embryos studied included uniform haploids after extrusion of the second polar body, mosaic haploids in which each blastomere contained a genetically different haploid nucleus, and heterozygous diploid mouse embryos. Eggs were activated and cultured in a chemically denned medium. About three times as many mosaic haploid or heterozygous diploid eggs developed beyond the 4-cell stage after 98–100 h and to the blastocyst stage after 120 h in culture, than uniform haploid eggs. This indicates that the development of parthenogenetic embryos is probably under genetic control and that there was a better development of the heterozygous embryos. Mosaic haploid embryos showed the same high frequency of development as heterozygous diploids. The results therefore indicate that heterozygosity provided a developmental advantage even when distributed between two genetically different clones of cells in the same embryo.

Cell division and cell allocation in early mouse development

Development ◽  
1978 ◽  
Vol 48 (1) ◽  
pp. 37-51
Author(s):  
S. J. Kelly ◽  
J. G. Mulnard ◽  
C. F. Graham
Keyword(s):  
Cell Division ◽  
Tritiated Thymidine ◽  
Mouse Embryos ◽  
Mouse Development ◽  
Stages Of Development ◽  
Cell Stage ◽  
Cell Cycles ◽  
Short Cell ◽  
Early Mouse

Cell division was observed in intact and dissociated mouse embryos between the 2-cell stage and the blastocyst in embryos developing in culture. Division to the 4-cell stage was usually asynchronous. The first cell to divide to the 4-cell stage produced descendants which tended to divide ahead of those cells produced by its slow partner at all subsequent stages of development up to the blastocyte stage. The descendants of the first cell to divide to the 4-cell stage did not subsequently have short cell cycles. The first cell or last cell to divide from the 4-cell stage was labelled with tritiated thymidine. The embryo was reassembled, and it was found that the first pair of cells to reach the 8-cell stage contributed disproportionately more descendants to the ICM when compared with the last cell to divide to the 8-cell stage.

scholarly journals AGS3-dependent trans-Golgi network membrane trafficking is essential for compaction in mouse embryos

2020 ◽  
Vol 133 (23) ◽  
pp. jcs243238
Author(s):  
Zheng-Wen Nie ◽  
Ying-Jie Niu ◽  
Wenjun Zhou ◽  
Dong-Jie Zhou ◽  
Ju-Yeon Kim ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Trafficking ◽  
Fluorescent Protein ◽  
Mouse Embryos ◽  
Cell Contact ◽  
Protein Signaling ◽  
Cell Stage ◽  
Trans Golgi Network ◽  
Early Mouse ◽  
Golgi Network

ABSTRACTActivator of G-protein signaling 3 (AGS3, also known as GPSM1) regulates the trans-Golgi network. The AGS3 GoLoco motif binds to Gαi and thereby regulates the transport of proteins to the plasma membrane. Compaction of early embryos is based on the accumulation of E-cadherin (Cdh1) at cell-contacted membranes. However, how AGS3 regulates the transport of Cdh1 to the plasma membrane remains undetermined. To investigate this, AGS3 was knocked out using the Cas9-sgRNA system. Both trans-Golgi network protein 46 (TGN46, also known as TGOLN2) and transmembrane p24-trafficking protein 7 (TMED7) were tracked in early mouse embryos by tagging these proteins with a fluorescent protein label. We observed that the majority of the AGS3-edited embryos were developmentally arrested and were fragmented after the four-cell stage, exhibiting decreased accumulation of Cdh1 at the membrane. The trans-Golgi network and TMED7-positive vesicles were also dispersed and were not polarized near the membrane. Additionally, increased Gαi1 (encoded by GNAI1) expression could rescue AGS3-overexpressed embryos. In conclusion, AGS3 reinforces the dynamics of the trans-Golgi network and the transport of TMED7-positive cargo containing Cdh1 to the cell-contact surface during early mouse embryo development.

Cell membrane regions in preimplantation mouse embryos

Development ◽  
1980 ◽  
Vol 59 (1) ◽  
pp. 89-102
Author(s):  
L. Izquierdo ◽  
T. López ◽  
P. Marticorena
Keyword(s):  
Enzyme Activity ◽  
Cell Membrane ◽  
Light And Electron Microscopy ◽  
Enzyme Reaction ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Cortical System ◽  
Crystalloid Bodies

Cell membrane regions characterized by alkaline phosphatase activity are described in cleaving mouse embryos and early blastocysts. Enzyme activity is demonstrated by light and electron microscopy, from the late 4-cell stage onwards, on the cell surfaces between blastomeres but not on the outer surface of the embryo. Experiments with dissociated morulae show that this is probably not an artifact due to the retention of the enzyme reaction product between the blastomeres. With the electron microscope the activity is also demonstrated in crystalloid bodies within the cytoplasm. The localization and growth during cleavage of cell membrane regions with enzyme activity is interpreted as the result of new cell membrane formation and/or as a relation of the crystalloid bodies with the cell membrane through the cortical system of microtubules and filaments.

Nuclei from fertilized mouse embryos have calcium-releasing activity

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1123-1128 ◽  
Author(s):  
T. Kono ◽  
J. Carroll ◽  
K. Swann ◽  
D.G. Whittingham
Keyword(s):  
Nuclear Transfer ◽  
Mouse Embryos ◽  
Oocyte Activation ◽  
Cell Stage ◽  
Nuclear Envelope Breakdown ◽  
Mammalian Embryos ◽  
Sperm Factor ◽  
Physical Stimuli ◽  
Mammalian Fertilization ◽  
Parthenogenetic Embryos

During mammalian fertilization, the sperm triggers a series of intracellular Ca2+ oscillations which initiate oocyte activation and the formation of pronuclei. Oocyte activation can be induced artificially by a variety of chemical and physical stimuli which elevate intracellular calcium. We show that the transfer of nuclei from 1- and 2-cell-stage fertilized mouse embryos to unfertilized oocytes stimulates the completion of meiosis and the formation of pronuclei. Nuclei from embryos that had developed to the 4-cell stage did not stimulate meiotic resumption. The ability to cause oocyte activation was specific to nuclei transferred from fertilized embryos as nuclei from parthenogenetic embryos or cytoplasts from fertilized or parthenogenetic embryos did not induce activation. Nucleus-induced oocyte activation was associated with the generation of intracellular Ca2+ transients, which were seen after nuclear envelope breakdown of the transferred nuclei. Treatment of the oocyte with the intracellular Ca2+ chelator, BAPTA, prior to nuclear transfer inhibited intracellular Ca2+ transients and oocyte activation. The specific Ca(2+)-releasing activity of the nucleus was not caused by sperm-induced protein synthesis since similar activity was present in nuclei originating from embryos exposed to cycloheximide throughout fertilization. The specific ability of nuclei from fertilized embryos to stimulate Ca2+ transients and oocyte activation was also found in nuclei from embryos parthenogenetically activated by the injection of a partially purified cytosolic sperm factor. The results suggest that the fertilizing sperm introduces Ca(2+)-releasing activity which becomes associated with the nucleus of early mammalian embryos.

scholarly journals Clathrin-mediated endocytosis is essential for the selective degradation of maternal membrane proteins and preimplantation development

Development ◽  
2021 ◽  
Vol 148 (14) ◽  
Author(s):  
Akihito Morita ◽  
Yuhkoh Satouh ◽  
Hidetaka Kosako ◽  
Hisae Kobayashi ◽  
Akira Iwase ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Ubiquitin Proteasome System ◽  
Maternal Plasma ◽  
Early Embryogenesis ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Selective Degradation ◽  
Kinase C ◽  
Ubiquitin Proteasome

ABSTRACT Fertilization triggers significant cellular remodeling through the oocyte-to-embryo transition. In this transition, the ubiquitin-proteasome system and autophagy are essential for the degradation of maternal components; however, the significance of degradation of cell surface components remains unknown. In this study, we show that multiple maternal plasma membrane proteins, such as the glycine transporter GlyT1a, are selectively internalized from the plasma membrane to endosomes in mouse embryos by the late two-cell stage and then transported to lysosomes for degradation at the later stages. During this process, large amounts of ubiquitylated proteins accumulated on endosomes. Furthermore, the degradation of GlyT1a with mutations in potential ubiquitylation sites was delayed, suggesting that ubiquitylation may be involved in GlyT1a degradation. The clathrin inhibitor blocked GlyT1a internalization. Strikingly, the protein kinase C (PKC) activator triggered the heterochronic internalization of GlyT1a; the PKC inhibitor markedly blocked GlyT1a endocytosis. Lastly, clathrin inhibition completely blocked embryogenesis at the two-cell stage and inhibited cell division after the four-cell stage. These findings demonstrate that PKC-dependent clathrin-mediated endocytosis is essential for the selective degradation of maternal membrane proteins during oocyte-to-embryo transition and early embryogenesis.

Cell membrane regionalization in early mouse embryos as demonstrated by 5'-nucleotidase activity

Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 115-126
Author(s):  
L. Izquierdo ◽  
C. Ebensperger
Keyword(s):  
Enzyme Activity ◽  
Cell Membrane ◽  
External Surface ◽  
Enzyme Reaction ◽  
Mouse Embryos ◽  
Nucleotidase Activity ◽  
Cell Stage ◽  
Morula Stage ◽  
Selective Retention ◽  
Early Mouse

The distribution of 5'-nucleotidase activity in pre-implantation mouse embryos is studied by means ofa cytochemical method adapted from Uusitalo & Karnovsky (1977). The enzyme activity is detected, from the4-cell stage up to the morula stage, on discrete patches of the cell membane between blastomeres. Appropriatecontrols show that this distribution is not a localization artifact due to selective retention of the enzyme reaction product in the narrow interblastomeric spaces. In early blastocysts, as the blastocoel expands the enzyme activity on its lining disappears. The external surface of the trophectoderm in early blastocysts lacks any enzyme activity, whereas in late blastocysts a strong enzyme activity is detected at the embryonic trophectoderm, decreasing in intensity towards the opposite pole of the embryo. These results are compared to previous observations by other authors and the differences are mainly ascribed to differences in the cytochemical procedure employed. We conclude that during cleavage a gradual cell membrane regionalization unfolds, revealing a pattern that may be related to morphogenesis; in particular, to the localization of zonular tight junctions around the peripheral blastomeres of the morula (Izquierdo, 1977; Izquierdo, López & Marticorena, 1980).

Methylation levels of maternal and paternal genomes during preimplantation development

Development ◽  
1991 ◽  
Vol 113 (1) ◽  
pp. 119-127 ◽  
Author(s):  
S.K. Howlett ◽  
W. Reik
Keyword(s):  
Methylation Status ◽  
Germinal Vesicle ◽  
Preimplantation Development ◽  
Preimplantation Embryos ◽  
Cell Stage ◽  
Global Methylation ◽  
Cell Cycles ◽  
Dna Methylase ◽  
Parthenogenetic Embryos ◽  
Metaphase Ii Oocytes

The methylation status of three highly repeated sequences was studied in sperm, eggs and preimplantation embryos with different combinations of parental chromosomes. High levels of methylation of the IAP and MUP sequence families were found in sperm and in eggs, whereas the L1 repeat was found to be highly methylated in sperm but only about 42% methylated in eggs. To assess how the two parental genomes behaved during preimplantation development, normal, fertilised embryos were compared with parthenogenetic embryos where the chromosomes are exclusively of maternal origin. It was observed that the high levels of methylation at the IAP and MUP sequences were retained through early development, with the first signs of demethylation at the IAP sequences apparent on both parental chromosomes in the blastocyst. Methylation at the sperm-derived L1 sequences dropped to about the same level as that of the egg-derived sequences by the late 2-cell stage, both then remain at this intermediate level until around the time of cavitation when levels fell to about 10% in the blastocyst. High levels of DNA methylase were detected in germinal vesicle and metaphase II oocytes; these high levels were maintained in fertilised and parthenogenetic embryos through into the morula and then declined to be undetectable in the blastocyst. Our comparison of maternal and paternal genomes suggests that methylation levels at repeat sequences are remarkably similar at the time of fertilisation or, as in the case of the L1 sequences, they become so during the first few cell cycles. Hence, there do not appear to be global methylation differences between the genomes that are retained through preimplantation development.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromosome analysis of early postimplantation presumptive haploid parthenogenetic mouse embryos

Development ◽  
1978 ◽  
Vol 45 (1) ◽  
pp. 85-91
Author(s):  
Matthew H. Kaufman
Keyword(s):  
Chromosome Analysis ◽  
Mouse Embryos ◽  
Chromosome Constitution ◽  
Equal Proportion ◽  
Dividing Cells ◽  
Parthenogenetic Mouse Embryos ◽  
Parthenogenetic Embryos ◽  
Apparently Healthy

The chromosome constitution of early postimplantation presumptive haploid parthenogenetic mouse embryos was examined. All the embryos isolated were at the egg-cylinder stage and seven contained dividing cells. In two of the apparently healthy embryos only haploid mitoses were seen, whereas in five others an approximately equal proportion of haploid and diploid mitoses was observed. Out of 52 cells in which unequivocal counts could be made, only one contained more than the euploid number of chromosomes (mouse, n = 20). Possible reasons for the poorer development of haploid compared to diploid parthenogenetic embryos are discussed.

Electron microscopy of endocardial cell populations

1978 ◽  
Vol 36 (2) ◽  
pp. 486-487
Author(s):  
T. G. Sarphie ◽  
C. R. Comer ◽  
D. J. Allen
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Cellular Transformation ◽  
Cell Populations ◽  
Cell Surfaces ◽  
Kb Cells ◽  
Dna Viruses ◽  
Cell Cycles ◽  
Ultrastructural Studies ◽  
Endocardial Cell

Previous ultrastructural studies have characterized surface morphology during norma cell cycles in an attempt to associate specific changes with specific metabolic processes occurring within the cell. It is now known that during the synthetic ("S") stage of the cycle, when DNA and other nuclear components are synthesized, a cel undergoes a doubling in volume that is accompanied by an increase in surface area whereby its plasma membrane is elaborated into a variety of processes originally referred to as microvilli. In addition, changes in the normal distribution of glycoproteins and polysaccharides derived from cell surfaces have been reported as depreciating after cellular transformation by RNA or DNA viruses and have been associated with the state of growth, irregardless of the rate of proliferation. More specifically, examination of the surface carbohydrate content of synchronous KB cells were shown to be markedly reduced as the cell population approached division Comparison of hamster kidney fibroblasts inhibited by vinblastin sulfate while in metaphase with those not in metaphase demonstrated an appreciable decrease in surface carbohydrate in the former.

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