Vol 48, No 4 (2017)

The presence of a complex structured brain, the anterior department of which (telencephalon) has no analogues in other groups of animals, is a significant peculiarity of vertebrates (including the human). Previously, a vertebrate-specific monogenic class of homeobox genes Anf, which plays a key role in the development of telencephalon, was discovered in the Laboratory of Molecular Bases of Embryogenesis (Institute of Bioorganic Chemistry, Russian Academy of Sciences). Out of all vertebrates, the Anf genes were not found only in the members of the most ancient group of jawless fishes (modern lampreys and hagfish), while the telencephalon was described in these animals. According to the literature data, a branch leading to jawless fishes separated from common vertebrate stem at the earliest stages of the their evolution, and detection of the Anf genes in jawless fishes is important, since it could confirm the previous hypothesis that the origin of telencephalon in evolution was associated with the appearance of the homeobox Anf gene. In the present work, the Anf gene was for the first time described in the Pacific lamprey (Lethenteron camtschaticum), the pattern and dynamics of its expression were studied at early stages of the development of this lamprey species, and the translation of its full-size cDNA was conducted in a eukaryotic expression system (clawed frog embryos).

Energy metabolism is studied in great ramshorn Planorbarius corneus during embryonic development. It is shown that the rate of oxygen consumption is constantly increasing in the process of embryogenesis. The respiration intensity (rate of the oxygen consumption per unit of the embryo volume) initially increases and then slowly decreases until eclosion. At the early stages of development until the early trochophore stage, the embryo is not growing, and, thus, the change in the rate of oxygen consumption during this period is not associated with the change of the embryo volume. Reduction in the intensity of respiration begins simultaneously with the beginning of the growth of the embryo at the stage of the middle trochophore. Starting from the middle trochophore and until eclosion, an association between oxygen consumption rate and volume of the embryo can be described with an allometric equation with exponential coefficient equal to approximately 0.23.

The naphthoquinone pigments of epidermal cells and gametes of clypeasteroid sand dollar Scaphechinus mirabilis isolated enzymatically with subsequent alcohol extraction were studied. It was found that naphthoquinone pigments are present in the pigment cells incorporated into the jelly coat of oocytes but not in the cytoplasm of unfertilized eggs. Laser desorption/ionization mass spectra showed that the pigment cells incorporated into the coats of mature egg cells contain spinochromes E and D, and those found in the epidermis of adult urchins contain echinochrome A and spinochrome D. No spinochromes were found in eggs lacking coats. Fertilization of sea urchins is accompanied by oxidative burst associated with the production of hydrogen peroxide from molecular oxygen by quinone- and naphthoquinone-dependent oxidase Udx1. Since there were no quinones in the egg cells of S. mirabilis, it can be assumed that water-soluble spinochrome E, penetrating by diffusion into the egg cytoplasm from the pigment cells of the coat, is used for hydrogen peroxide stimulation of embryogenesis. Echinochrome A, the dominant echinochrome in epidermal cells of the adult urchin, is insoluble in water and, apparently, immobilized by the ethyl group as a hydrophobic anchor.

The aim of this study was to analyze the changes that occur in the population of bone marrow mesenchymal stromal cells (MSCs) during the individual development of an organism. For this purpose, the basic characteristics of MSCs (the content of clonogenic cells, immunophenotype, and potencies to differentiate in vitro and in vivo) in the prenatal, early postnatal, and late postnatal ontogeny of the rat were compared. It is shown that the cloning efficiency of bone marrow MSCs in 10-day-old and adult rats is comparable and hundreds of times smaller than that of bone cells of 20-day-old fetuses with a bone marrow rudiment. The activity of alkaline phosphatase, a marker of osteogenic cells, was found in the majority of colonies formed by MSCs of postnatal bone marrow but not by the fetal bone. By the CD90 expression and potencies for in vitro adipogenesis, the stromal cells from the fetal bone and bone marrow of 9- to 10-day-old rats were comparable with those of the mature bone marrow MSCs but differed from them by the small number of CD73-bearing cells and a weaker ability to osteogenesis in an inductive environment. The analysis of the fate of MSCs from the studied sources after their transplantation to adult rats showed that their ectopic transplantation as part of tissue fragments into the kidney results in the formation of bone tissues and hematopoietic stroma. In diffusion chambers with MSCs that were precultured in vitro, transplantation into the peritoneal cavity led to osteogenesis and chondrogenesis. However, no significant differences in the potencies of bone marrow MSCs for differentiation in vivo depending on the developmental stage have been found. Thus, during ontogeny, bone marrow MSCs enhance the expression of CD73 and the ability to osteogenesis in vitro, whereas the expression of CD90 and the potencies for adipogenesis in induction medium and differentiation in different directions in vivo do not change significantly.

Using immunohistochemical methods, the morphological features of the sympathetic nerve structures in the pancreas of newborn, pubescent, and aging rats have been studied. The neural composition of intramural ganglia has been described. The intramural ganglia were shown to include chromaffin cells. In many ganglia of the pancreas, two types of pericellular nerve apparatuses have been detected simultaneously: tyrosine hydroxylase-containing catecholaminergic synaptic terminals and PGP 9.5-immunopositive cholinergic synapses. It was established that the density of catecholaminergic structures in the pancreas of rats decreases with age.

Cancer-testis antigens of the Mage family (Melanoma antigens) are expressed predominantly in the spermatogenic and cancer cells, but some genes of this family are expressed ubiquitously. Expression patterns and functional role of Mage family antigens in the regulation of cellular processes in normal embryonic and definitive cells are virtually unknown. Comparative immunofluorescent analysis of Mage expression in mouse oocytes and early embryos identified the expression of Mage antigens at all stages studied. The greatest intensity of the fluorescent staining was detected in the epiblasts and the extraembryonic structures of the egg cylinder at E6.5 stage. At all studied developmental stages of the mouse oocyte and the early embryo, the localization of Mage antigens was found predominantly in the cytoplasm. Quantitative real-time PCR showed that expression levels of most Mage genes in cells of the epiblast and ectoplacental cone were similar, while the gene expression levels of Mage-a10, Mage-b16, and Mage-b18 were higher in cells of the ectoplacental cone than in epiblast cells. Thus, for the first time, our analysis has shown that the Mage family antigens are expressed at the early stages of mouse development and may be involved in the regulation of earliest events of embryogenesis.