The DDK inbred strain as a model for the study of interactions between parental genomes and egg cytoplasm in mouse preimplantation development

Development ◽  
1990 ◽  
Vol 108 (Supplement) ◽  
pp. 81-87 ◽  
Author(s):  
Charles Babinet ◽  
Véronique Richoux ◽  
Jean-Louis Guénet ◽  
Jean-Paul Renard
Keyword(s):  
Specific Interaction ◽  
Blastocyst Stage ◽  
Low Fertility ◽  
Nuclear Transplantation ◽  
Diploid Nucleus ◽  
Early Embryonic Lethality ◽  
Molecular Studies ◽  
Per Se ◽  
Transplantation Experiments ◽  
Nucleocytoplasmic Interactions

The DDK strain of mice has unusual genetic properties. When females of this strain are crossed to males of other strains, they generally exhibit a very low fertility, whereas reciprocal crosses are fully fertile as are the intrastrain crosses. The observed low fertility results from early embryonic lethality, the F1 embryos dying around the late morula–early blastocyst stage. Nuclear transplantation experiments between hybrid eggs of BALB/c and DDK strains has shown that failure of F1(DDK ♀× BALB/c♂) embryos to develop is not due to the combination per se of maternal (DDK) and paternal (BALB/c) genomes but rather to an incompatibility between paternal (BALB/c) genomic contribution and DDK cytoplasm. This incompatibility does not occur between a female BALB/c pronucleus and the DDK cytoplasm, suggesting the involvement of a differential imprinting of parental genomes. Introduction of cytoplasts isolated from DDK 1- to 8-cell embryos into BALB/c♀×BALB/c♂ or BALB/c♀×DDK♂ embryos of the corresponding developmental stage demonstrate that the cytoplasm of DDK embryos prevents the formation of normal blastocysts through a specific interaction with the paternal component of the BALB/c diploid nucleus. Genetic and molecular studies are underway to try and isolate the gene(s) responsible for the failure of (DDK♀×BALB/c♂)F1 embryos. These experiments should help in our understanding of nucleocytoplasmic interactions and the respective roles of parental genomes in early embryonic development.

scholarly journals Genetic and Molecular Studies on Om, a Locus Controlling Mouse Preimplantation Development

1996 ◽  
Vol 45 (1-2) ◽  
pp. 3-14 ◽  
Author(s):  
M. Cohen-Tannoudj ◽  
P. Balducci ◽  
C. Kress ◽  
V. Richoux-Duranthon ◽  
J.P. Renard ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Female Parent ◽  
Blastocyst Stage ◽  
Nuclear Transplantation ◽  
Genetic Constitution ◽  
Cell Nuclei ◽  
Cell Stage ◽  
Female Pronucleus ◽  
Haploid Embryos ◽  
Transplantation Experiments

Several lines of evidence have accumulated in recent years indicating that nuclear cytoplasmic interactions play an important role in the formation and fate of the developing mouse embryo. Early nuclear transplantation experiments indicated that the ability of nuclei to direct cleavage after transfer into enucleated zygotes falls abruptly with nuclei from more advanced preimplantation stages [1]. Transcriptional activation of the nuclei, which occurs during the second cell cycle probably precludes the reprogramming of nuclei from later cleavage stages [2]. Thus, when an 8-cell nucleus is transferred to an enucleated zygote, such a reconstituted zygote is blocked at the 2-cell stage. However, when identical 8-cell nuclei were transferred into both blastomeres of enucleated 2-cell embryos, they were able to support development to the blastocyst stage and even gave rise to live offspring [2-4]. This indicated the importance of the cytoplasmic environment for the ability of the incoming nucleus to support development. It should be noted that in these experiments, the nuclear cytoplasmic ratio was also an important factor in determining the development of the reconstituted embryos [2]. Similar observations were also made when monitoring the development of haploid embryos [5]. In another study, Latham and Solter [6] examined the ability of androgenones, obtained by replacing the female pronucleus of a zygote by the male pronucleus, to develop to the blastocyst stage. Androgenones generated from C57B1/6 eggs were found to be much more competent to give rise to blastocysts than were DBA/2 androgenones. However, when androgenones were constructed from (DBA/2×C57B1/6)F1, zygotes (genetic constitution of the embryos will hereafter be indicated with the female parent coming first followed by the male parent), by replacing the DBA/2 female pronucleus with a C57B1/6 pronucleus, they also developed poorly. This was not simply due to the lack of some component in DBA/2 cytoplasm, since the impaired development was also observed when C57B1/6 male pronuclei from pairs of (DBA/2×C57B1/6) F1, were transferred to an enucleated C57B1/6 egg.

Ploidy, pigment patterns and species specific antigenicity in interspecific nuclear transplantations in newts

Development ◽  
1967 ◽  
Vol 17 (2) ◽  
pp. 319-330
Author(s):  
F. Sládeček ◽  
A. Romanovský
Keyword(s):  
Xenopus Laevis ◽  
Functional Activity ◽  
Nuclear Transplantation ◽  
Skin Grafts ◽  
Species Specific ◽  
Transplantation Antigens ◽  
Transplantation Experiments ◽  
Pigment Patterns

It was shown by Simnett (1964) that in Xenopus laevis skin grafts in adult frogs between members of the same nuclear clone were tolerated in the same way as autografts, but in skin grafts made between individuals belonging to different nuclear clones a homograft rejection occurred. The nucleus is therefore responsible for the synthesis of specific transplantation antigens. It seemed to us useful to investigate the species-specific antigenicity of animals derived from eggs transplanted with foreign nuclei in correlation with their ploidy and with the development of their species-specific pigment patterns, as a proof of functional activity of transplanted nuclei. For this purpose we used two species of Triturus, T. vulgaris and T. alpestris, because of earlier studies carried out in our laboratory on the pigmentation of their hybrids (Romanovský & Ŝtefanová, 960; Mazáková-Štefanová, 1965) and on their species-specific antigenicity (Romanovský, 1962 a, b), in spite of the known difficulties and limitations of nuclear transplantation experiments in these species (Lehman, 1955; Sládeček & Mazáková-Štefanová, 1964, 1965).

Nuclear transplantation of male primordial germ cells in the mouse

Development ◽  
1989 ◽  
Vol 107 (2) ◽  
pp. 407-411 ◽  
Author(s):  
Y. Tsunoda ◽  
T. Tokunaga ◽  
H. Imai ◽  
T. Uchida
Keyword(s):  
Germ Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Nuclear Transplantation ◽  
Embryonic Genome ◽  
Donor Nucleus ◽  
Implantation Sites ◽  
Genome Activation

We examined the developmental ability of enucleated eggs receiving embryonic nuclei and male primordial germ cells (PGCs) in the mouse. Reconstituted eggs developed into the blastocyst stage only when an earlier 2-cell nucleus was transplanted (36%) but very rarely if the donor nucleus was derived from a later 2-cell, 8-cell, or inner cell mass of a blastocyst (0–3%). 54–100%, 11–67%, 6–43% and 6–20% of enucleated eggs receiving male PGCs developed to 2-cell, 4-cell, 8-cell and blastocyst stage, respectively, in culture. The overall success rate when taking into account the total number of attempts at introducing germ cells was actually 0–6%. Live fetuses were not obtained after transfer of reconstituted eggs to recipients, although implantation sites were observed. The developmental ability of reconstituted eggs in relation to embryonic genome activation and genomic imprinting is discussed.

scholarly journals PROTEINS IN NUCLEOCYTOPLASMIC INTERACTIONS

1968 ◽  
Vol 39 (2) ◽  
pp. 404-414 ◽  
Author(s):  
David Prescott ◽  
Lester Goldstein
Keyword(s):  
Binding Sites ◽  
Interphase Nucleus ◽  
Nuclear Protein ◽  
Protein A ◽  
Amoeba Proteus ◽  
Nuclear Proteins ◽  
Nuclear Transplantation ◽  
Amino Acid Labeling ◽  
Turn Over ◽  
Nucleocytoplasmic Interactions

The behavior of nuclear proteins in Amoeba proteus was studied by tritiated amino acid labeling, nuclear transplantation, and cytoplasmic amputation. During prophase at least 77% (but probably over 95%) of the nuclear proteins is released to the cytoplasm. These same proteins return to the nucleus within the first 3 hr of interphase. When cytoplasm is amputated from an ameba in mitosis (shen the nuclear proteins are in the cytoplasm), the resultant daughter nuclei are depleted in the labeled nuclear proteins. The degree of depletion is less than proportional to the amount of cytoplasm removed because a portion of rapidly migrating protein (a nuclear protein that is normally shuttling between nucleus and cytoplasm and is thus also present in the cytoplasm) which would normally remain in the cytoplasm is taken up by the reconstituting daughter nuclei. Cytoplasmic fragments cut from mitotic cells are enriched in both major classes of nuclear proteins, i.e. rapidly migrating protein and slow turn-over protein. An interphase nucleus implanted into such an enucleated cell acquires from the cytoplasm essentially all of the excess nuclear proteins of both classes. The data indicate that there is a lack of binding sites in the cytoplasm for the rapidly migrating nuclear protein. The quantitative aspects of the distribution of rapidly migrating protein between the nucleus and the cytoplasm indicate that the distribution is governed primarily by factors within the nucleus.

An overview of glossiphoniid leech development

2001 ◽  
Vol 79 (2) ◽  
pp. 218-232 ◽  
Author(s):  
David A Weisblat ◽  
Françoise Z Huang
Keyword(s):  
Model Systems ◽  
Model Organisms ◽  
Consensus Tree ◽  
Comparative Development ◽  
Experimental Embryology ◽  
Molecular Studies ◽  
Research Questions ◽  
Per Se ◽  
Molecular Phylogenies ◽  
Development And Evolution

Dramatic advances in understanding the development of selected "model" organisms, coupled with the realization that genes which regulate development are often conserved between diverse taxa, have renewed interest in comparative development and evolution. Recent molecular phylogenies seem to be converging on a new consensus "tree," according to which higher bilaterians fall into three major groups, Deuterostoma, Ecdysozoa, and Lophotrochozoa. Commonly studied model systems for development fall almost exclusively within the first two of these groups. Glossiphoniid leeches (phylum Annelida) offer certain advantages for descriptive and experimental embryology per se, and can also serve to represent the lophotrochozoan clade. We present an overview of the development of glossiphoniid leeches, highlighting some current research questions and the potential for comparative cellular and molecular studies.

scholarly journals CRISPR/Cas9-mediated knock-out of dUTPase in mice leads to early embryonic lethality

2018 ◽  
Author(s):  
Hajnalka Laura Pálinkás ◽  
Gergely Rácz ◽  
Zoltán Gál ◽  
Orsolya Hoffmann ◽  
Gergely Tihanyi ◽  
...  
Keyword(s):  
De Novo ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Genome Maintenance ◽  
Inorganic Pyrophosphate ◽  
Knock Out ◽  
Early Embryonic Lethality ◽  
Unicellular Organisms ◽  
Post Implantation

AbstractSanitization of nucleotide pools is essential for genome maintenance. Among the enzymes significant in this mechanism, deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) performs cleavage of dUTP into dUMP and inorganic pyrophosphate. By this reaction the enzyme efficiently prevents uracil incorporation into DNA and provides dUMP, the substrate for de novo thymidylate biosynthesis. Despite its physiological significance, knock-out models of dUTPase have not yet been investigated in mammals, only in unicellular organisms, such as bacteria and yeast. Here we generate CRISPR/Cas9-mediated dUTPase knock-out in mice. We find that heterozygous dut +/-animals are viable while the decreased dUTPase level is clearly observable. We also show that the enzyme is essential for embryonic development. Based on the present results, early dut -/-embryos can still reach the blastocyst stage, however, they die shortly after implantation. Analysis of preimplantion embryos indicate perturbed growth of both inner cell mass (ICM) and trophectoderm (TE). We conclude that dUTPase is indispensable for post-implantation development in mice. The gene targeting model generated in the present study will allow further detailed studies in combination with additional gene knock-outs.

scholarly journals Direct somatic lineage conversion

2015 ◽  
Vol 370 (1680) ◽  
pp. 20140368 ◽  
Author(s):  
Koji Tanabe ◽  
Daniel Haag ◽  
Marius Wernig
Keyword(s):  
Transcription Factors ◽  
Cell Types ◽  
Nuclear Transplantation ◽  
Specific Cell ◽  
Direct Reprogramming ◽  
Cell Type ◽  
Mesodermal Cell ◽  
Related Cell ◽  
Transplantation Experiments ◽  
Cell Specialization

The predominant view of embryonic development and cell differentiation has been that rigid and even irreversible epigenetic marks are laid down along the path of cell specialization ensuring the proper silencing of unrelated lineage programmes. This model made the prediction that specialized cell types are stable and cannot be redirected into other lineages. Accordingly, early attempts to change the identity of somatic cells had little success and was limited to conversions between closely related cell types. Nuclear transplantation experiments demonstrated, however, that specialized cells even from adult mammals can be reprogrammed into a totipotent state. The discovery that a small combination of transcription factors can reprogramme cells to pluripotency without the need of oocytes further supported the view that these epigenetic barriers can be overcome much easier than assumed, but the extent of this flexibility was still unclear. When we showed that a differentiated mesodermal cell can be directly converted to a differentiated ectodermal cell without a pluripotent intermediate, it was suggested that in principle any cell type could be converted into any other cell type. Indeed, the work of several groups in recent years has provided many more examples of direct somatic lineage conversions. Today, the question is not anymore whether a specific cell type can be generated by direct reprogramming but how it can be induced.

Nuclear transplantation using bovine primordial germ cells from male fetuses

1995 ◽  
Vol 7 (5) ◽  
pp. 1217 ◽  
Author(s):  
F Delhaise ◽  
FJ Ectors ◽  
Roover R de ◽  
F Ectors ◽  
F Dessy
Keyword(s):  
Nuclear Transfer ◽  
Primordial Germ Cells ◽  
Blastocyst Stage ◽  
Cell Counting ◽  
Nuclear Transplantation ◽  
Cell Nuclei ◽  
Cell Stage ◽  
Developmental Potential ◽  
Morula Stage

The developmental potential of nuclei of bovine gonial cells was investigated by nuclear transfer. Gonial cells were collected from male fetuses at about 175 days post coitum (p.c.). They were fused with enucleated oocytes; reconstituted embryos were cultured in vitro for 7 days. Embryos reaching the compacted morula or blastocyst stage were either fixed for cell counting or transferred into recipients. Out of 115 oocyte-gonia fusions, 101 (87.8%) gave rise to cleaved embryos at Day 3 and 26 (22.6%) had reached the 8-cell stage. At Day 7, 1 (1%) developed to the morula stage and 5 (4%) reached the blastocyst stage. Three blastocysts were fixed and showed normal cell numbers (135; 90; 76 cells). Three blastocysts and one morula were transferred in four recipients; two recipients were pregnant at Day 21 but only one was positive at Day 35 p.c.; this last one aborted around Day 40 p.c. No conceptus was collected. These results indicate that gonial cell nuclei can be partially reprogrammed; they are able to develop into blastocysts and to initiate gestation. However, more experiments will be necessary to prove the nuclear totipotency of bovine gonial cells.

scholarly journals CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality

Biomolecules ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 136 ◽  
Author(s):  
Hajnalka Pálinkás ◽  
Gergely Rácz ◽  
Zoltán Gál ◽  
Orsolya Hoffmann ◽  
Gergely Tihanyi ◽  
...  
Keyword(s):  
De Novo ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Genome Maintenance ◽  
Inorganic Pyrophosphate ◽  
Knock Out ◽  
Early Embryonic Lethality ◽  
Key Enzyme ◽  
Unicellular Organisms

Sanitization of nucleotide pools is essential for genome maintenance. Deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) is a key enzyme in this pathway since it catalyzes the cleavage of 2′-deoxyuridine 5′-triphosphate (dUTP) into 2′-deoxyuridine 5′-monophosphate (dUMP) and inorganic pyrophosphate. Through its action dUTPase efficiently prevents uracil misincorporation into DNA and at the same time provides dUMP, the substrate for de novo thymidylate biosynthesis. Despite its physiological significance, knock-out models of dUTPase have not yet been investigated in mammals, but only in unicellular organisms, such as bacteria and yeast. Here we generate CRISPR/Cas9-mediated dUTPase knock-out in mice. We find that heterozygous dut +/– animals are viable while having decreased dUTPase levels. Importantly, we show that dUTPase is essential for embryonic development since early dut −/− embryos reach the blastocyst stage, however, they die shortly after implantation. Analysis of pre-implantation embryos indicates perturbed growth of both inner cell mass (ICM) and trophectoderm (TE). We conclude that dUTPase is indispensable for post-implantation development in mice.

BALB/c Alleles at Modifier Loci Increase the Severity of the Maternal Effect of the “DDK Syndrome”

Genetics ◽  
2000 ◽  
Vol 154 (2) ◽  
pp. 803-811
Author(s):  
Stéphanie Le Bras ◽  
Michel Cohen-Tannoudji ◽  
Chantal Kress ◽  
Sandrine Vandormael-Pournin ◽  
Charles Babinet ◽  
...  
Keyword(s):  
Genetic Background ◽  
Reciprocal Cross ◽  
Inbred Mouse ◽  
Inbred Mouse Strain ◽  
Inbred Strains ◽  
Blastocyst Stage ◽  
Low Fertility ◽  
Intermediate Phenotype ◽  
Parental Effect ◽  
Genetic Studies

Abstract The Om locus was first described in the DDK inbred mouse strain: DDK mice carry a mutation at Om resulting in a parental effect lethality of F1 embryos. When DDK females are mated with males of other (non-DDK) inbred strains, e.g., BALB/c, they exhibit a low fertility, whereas the reciprocal cross, non-DDK females × DDK males, is fertile (as is the DDK intrastrain cross). The low fertility is due to the death of (DDK × non-DDK)F1 embryos at the late-morula to blastocyst stage, which is referred to as the “DDK syndrome.” The death of these F1 embryos is caused by an incompatibility between a DDK maternal factor and the non-DDK paternal pronucleus. Previous genetic studies showed that F1 mice have an intermediate phenotype compared to parental strains: crosses between F1 females and non-DDK males are semisterile, as are crosses between DDK females and F1 males. In the present studies, we have examined the properties of mice heterozygous for BALB/c and DDK Om alleles on an essentially BALB/c genetic background. Surprisingly, we found that the females are quasi-sterile when mated with BALB/c males and, thus, present a phenotype similar to DDK females. These results indicate that BALB/c alleles at modifier loci increase the severity of the DDK syndrome.

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