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The nucleus of mouse two-cell embryos houses the coilin-containing bodies of two types: 1) 1—3 large spherical structures of 1 mm and 2) small foci, which vary in number in different blastomeres. The largest coilin-containing structures, unlike the smallest ones, contain RNA polymerase I, nucleic acid chaperon YB-1, and also actin. Neither large nor small coilin-positive domains contain symplekin, one of the signature components of histone locus bodies. In the nuclei of late two-cell embryos, symplekin localizes to 1—2 well-formed roundish bodies that are observed both in close proximity to the coilin-positive structures and far away from them. Large coilin-containing bodies were not observed in embryos at the morula stage as well as in the nuclei of late two-cell embryos after artificial suppression of transcription activity. Thus, a population of coilin-containing bodies in the nuclei of late two-cell embryos of mice is heterogeneous in morphology and molecular composition. It could be assumed that the largest coilin-containing bodies are provisional nuclear domains that are formed at the background of significant changes of nuclear metabolism at the final stages of embryonic genome activation and the initial stages of reactivation of nucleolar transcription.

This paper presents a brief survey and preliminary classification of embryonic cleavage patterns in the class Amphibia. We use published data on 41 anuran and 22 urodele species concerning the character of the third cleavage furrow (latitudinal or longitudinal) and the stage of transition from synchronous to asynchronous blastomere divisions in the animal hemisphere (4-8-celled stage, 8-16-celled stage or later). Based on this, four patterns of amphibian embryonic cleavage are recognized, and an attempt to elucidate the evolutionary relationships among these patterns is undertaken. The so-called "standard" cleavage pattern (the extensive series of synchronous blastomere divisions including latitudinal furrows of the third cleavage) with the typical model species Ambystoma mexicanum and Xenopus laevis seems to be derived and probably originated independently in the orders Anura and Caudata. The ancestral amphibian cleavage pattern seems to be represented by species with longitudinal furrows of the third cleavage and the loss ofsynchrony as early as the 8-celled stage (such as in primitive urodele species from the family Cryptobranchidae).

In Spriralia, the specification of cell lines in the course of development is provided by maternal factors. However, recent studies demonstrated the importance of inductive processes whose significant element is cellular signaling. Our data allow us to speak of the dependent specification of a number of cell lines at the early stages of development of the mollusk Testudinalia testudinalis (Testudinalia tessellate, Patellogastropoda), including the period when the determination of the 3D cell takes place, which is accompanied by a change in the shape and establishing of contacts with animal micromeres by one of the macromeres of the third quartet. It is exactly at this moment that activation of MARK was registered in the 3D blastomere-organizer. An analysis of the influence of the U0126 blocker of the MAP-kinase way on the development of Testudinalia showed that the greatest effect of the inhibitor is observed during the cultivation of embryos until the sixth cycle of fragmentation. It should be noted that a scale of correlation of the degree of defects and increase in concentration exists. Absence of the functioning retractor, disorganization of the muscle system, and abnormal structure of the shell (to the extent of complete absence of the shell), as well as velum, foot, and mantle fold, were observed in a considerable part of larvae after a lengthy upkeep of the objects in the U0126 solution. At the same time, none of the experiments showed a complete disruption of the specification of the dorsoventral axis, which produces a larva with a four-ray radial symmetry. This attests in favor of the existence of various molecular mechanisms of determination of the secondary body axis among the animals from the group Spiralia.

Gap junctions (GJs) are composed of membrane proteins that form channels connecting the cytoplasm of adjacent cells and permeable to ions and small molecules. They are considered to be the main or only type of intercellular channels and a universal feature of all multicellular animals (Metazoa). Till recently, sea anemones and corals (Anthozoa, Cnidaria) appeared to be an exception from this rule. There were no structural or physiological data supporting the presence of GJ in Anthozoa. For some time no genes homologous to GJ proteins (connexins or pannexins) were detected in sea anemone Nematostella vectensis (Cnidaria, Anthozoa) or other Anthozoa genomes. Recently, pannexin homolog was found in Nematostella. Our intracellular recordings demonstrate electrical coupling between blastomeres in embryos at the 8-cells stage. At the same time, carboxyfluorescein fluorescent dye did not diffuse between electrically coupled cells, which excludes the possibility that the observed electrical coupling is mediated by incomplete cytoplasm separation during the cleavage. These data support the idea that GJ are ubiquitous for Metazoa, and pannexins are universal GJ proteins.

The changes in the state of long-term culture of blastocysts derived from female C57BL/6 mice after crossing with C57BL/6-Tg(ACTB-EGFP)1Osb/J males with a green fluorescent protein transgene EGFP on chromosome 15 were studied. Possible causes of different culture results were analyzed: the preservation of undifferentiated cells as dense clusters in the inner cell mass or their differentiation into extraembryonic endoderm. Comparison of the events going in blastocysts with the -/- or -/EGFP genotypes demonstrated that the GFP presence has no effect on cell processes. This allows us to use embryonic material from this mouse line in experiments that require long-term vital observation of embryonic cells.

Osmotic adaptation in a blastomere of mouse embryo has been studied by the direct measurement of the cell volume using laser scanning microscopy microtomography followed by quantitative 3D reconstruction. Embryonic cells subjected to hypotonic shock first swelled and then returned to the initial size. At the beginning of osmotic stress, the swelling occurred by the van't Hoff equation with the water permeability coefficient (L(p)) of 0.4 micro x min(-1) atm(-1). The phase of the regulatory volume decrease was not abolished by Na+/K(+)-ATPase inhibition.

The application of etoposide for chemical enucleation of rat oocytes was tested. The reconstruction efficiency after chemical and mechanical enucleation was comparatively analyzed. The obtained data indicate similar viability of reconstructed rat embryos irrespective of the enucleation technique.

Distribution of human mitochondrial DNA (mtDNA) among separate murine blastomeres was analyzed during the splitting of embryos in which the suspension of human mitochondria had been injected at the one- or two-cell stage. Human mtDNA was detected by PCR with species specific primers. The total amount of the two- and four-cell murine embryos analyzed in the study was 339. In all embryos examined the copies of human mitochondrial genome were revealed along with murine mtDNA, which indicated the phenomenon of an artificially modeled heteroplasmy. The foreign mtDNA was not ubiquitous among the blastomeres of transmitochondrial embryos. Mathematical analysis of the results showed that in the period between the injection of human mitochondria and the subsequent splitting no equal distribution of the human mtDNA occurred in the cytoplasm. These results also point at the presence of more than 2-3 segregation units of mtDNA in the entire pool of mitochondria (about 5 x 10(2)) introduced into an embryo by microinjection.

It was shown that local application of agonists of the 3rd type receptors SR57277A and quipazine into the interblastomere cleft of Paracentrotus lividus embryos evoked specific membrane currents. At the same time, ligands of 5-HT3-receptors specifically affected the cleavage patterns of half-embryos, i.e., imitated or avoided the interblastomere signal. In the view of the data obtained, we discuss a more precise concept of protosynapse, where the distribution of membrane serotonin receptors is restricted to the period of blastomere formation during cleavage and localized in the area of interblastomere contact.

We studied distribution of mRNA for nuclear protooncogene c-myc and nuclear protein P-53 in mature oocytes and embryos of Xenopus laevis from the stage of fertilization up to the stage of hatching by in situ hybridization with histological sections. mRNA for c-myc was present in all cells of the embryo at all studied developmental stages. Between the stage of fertilization and up to the late blastula, mRNA concentration for c-myc decreased progressively in all embryonic cells. During gastrulation a local increase in the concentration of this messenger was found in dorsal mesoderm and ectoderm. At the stage of neurula increased concentration of mRNA for c-myc was observed in all cells of the embryo but the hybridization signal increased particularly distinctly in cells of the neural tube. In studies of P-53 mRNA distribution hybridization signal was detected only in brain cells after stage 20 of development (after closure of the neural folds) and up to the stage of hatching.

Radioimmunological method was applied to the study of progesteron and 17 beta-estradiol profiles in the peripheral blood plasma of pregnant mice from the time of the zygote formation (the 1st day of gestation) to the blastocyst implantation (the 5th day of gestation). The peak of a progesteron concentration in the plasma and the onset of reduction in the 17 beta-estradiol concentration were noted in the evening of the 3rd day of pregnancy. From this period there was a fall in the 17 beta-estradiol and progesterone content in the blood plasma. In the evening of the 3rd day of pregnancy the 14-18-cell embryos passed from the oviduct into the uterine cornu. The role played by the embryo in changes of the maternal hormonal status is discussed.