Polytene chromosomes in mammalian cells.

This article deals with the structural and functional organization of polytene chromosomes in mammals. Based on cytophotometric, autoradiographic, and electron microscopic data, the authors put forward a concept of nonclassic polytene chromosomes, with special reference to polytene chromosomes in the mammalian placenta. In cells with nonclassic polytene chromosomes, two phases of the polytene nucleus cycle are described, such as the endointerphase (S phase) and endoprophase (G phase). The authors generalize that the main feature of nonclassic polytene chromosomes is that forces binding the sister chromatids are much weaker than in the Diptera classic polytene chromosomes. This concept is confirmed by comparative studies of human, mink, and fox polytene chromosomes. The final step of the trophoblast giant cell differentiation is characterized by a transition from polyteny to polyploidy, with subsequent fragmentation of the highly polyploid nucleus into fragments of low ploidy. Similarities and dissimilarities of pathways of formation and rearrangement of nonclassic polytene chromosomes in mammals, insects, plants, and protozoans are compared. The authors discuss the significance of polyteny as one of the intrinsic conditions for performance of the fixed genetic program of trophoblast giant cell development, a program that provides for the possibility of a long coexistence between maternal and fetal allogenic organisms during pregnancy.

[A study of DNA depolyploidization and depolytenization of the heterochromatized gonosomal chromatin bodies in the secondary giant trophoblast cells of the field vole Microtus rossiaemeridionalis using cytophotometry].

A study was made of the distribution of the heterochromatized gonosomal chromatin bodies (GCB) material in the course of nuclear fragmentation of secondary giant trophoblast cells resulting in polykaryocyte formation at the late stage of their differentiation. A simultaneous DNA cytophotometry in GCBs and nuclear fragments showed a progressive GCB DNA content decrease proportional to that of DNA content in nuclear fragments. DNA contents in the nuclear fragments corresponded to 2c, 4c and 8c. In most cases 1-2 GCBs were found in the nuclear fragments of different ploidy levels. Both the total DNA content in GCBs and the DNA content in separate GCBs well correlated with the ploidy levels of fragments. The data obtained demonstrate a regular, whole-genome distribution of chromosomal materials into the nuclear fragments exemplified by sex chromosome distribution in compliance with the ploidy of nuclear fragments. We discuss a possible mechanism of nuclear fragmentation that may ensure substantially a balanced genome of nuclear fragments without leading to mitotic cycle renewal in the giant trophoblast cell population.

Polyploidization in the trophoblast and uterine glandular epithelium of the endotheliochorial placenta of silver fox (Vulpes fulvus Desm.), as revealed by the DNA content.

Dynamics of genome multiplication during establishment of interrelations between trophoblast and glandular epithelium of the endometrium has been studied in the course of formation of placenta in the silver fox. During formation of the placenta, penetration of the trophoblast into the zone of the endometrial glandular epithelium and of endometrial blood vessels into the zone of expanding trophoblast occurs. The trophoblast, which gradually replaces epithelium and a part of the stroma of the endometrium, closely adjoins endometrial vessels but does not disrupt them, thereby the endotheliochorial placenta is formed. Cytophotometric measurements of the DNA content in trophoblast nuclei have shown that most of them are polyploid: predominantly 4-64c, occasionally 128c and 256c. Polyploidy of the trophoblast may be a consequence of various types of polyploidizing mitoses. Cytophotometric measurements of the DNA content in mitotic figures have revealed the presence of mitoses of diploid cells, i.e. with the DNA amount of 4c (2n), and polyploid cells, i.e. 8c (4n), and 16c (8n), therefore trophoblast cells in the silver fox placenta are able to enter mitosis up to the octaploid level. Higher degrees of polyploidy in the trophoblast cells seem to be achieved by endoreduplication. Polyploidization of the uterine glandular epithelial cells during placentation in the silver fox occurs until the level of 8c. Thus, the tissue-specific response of the uterus to the implanting embryo consists of active proliferation and polyploidization of the glandular epithelium, which may compensate formation of prominent population of decidual cells (i.e., connective tissue cells). In the endotheliochorial placenta of the silver fox the regularity is confirmed that cells of both maternal and fetal origin are, as a rule, polyploid in sites of their contact in placenta, which may be of protective significance in the contact of allogenic organisms.

[Glycogen-containing cells in the maternal and embryonic portions of the placenta in the rat and the common vole Microtus subarvalis]. / Glikogensoderzhashchie kletki v materinskoi i zarodysevoi chasti platsenty krysy i seroi polevki Microtus subarvalis.

Differentiation sequences and further transfiguration of glycogen-rich cells during placenta development were investigated for the rat and field vole Microtus subarvalis (11-20 day gestation). The presence of glycogen is a characteristic feature of decidual cells located in the region of lateral sinusoids, as well as of metrial gland cells, secondary giant trophoblast cells and trophoblast cells in the connective zone of placenta. Glycogen-containing metrial gland cells and trophoblast cells of connective zone of placenta are found to underlie the layer of tertiary giant trophoblast cells that cover the wall of the central arteria. Thus, both maternal and embryo-derived glycogen-containing cells always accompany the tertiary giant trophoblast cells that penetrate deeply into the maternal part of placenta but do not contain glycogen. In the field vole placenta the cells of peripheral trophoblast subpopulation of the connective zone of placenta attaching to the decidua basalis are stained by PAS-reaction more intensely than deeply situated ones. These data, as well as other phenomena revealed here, show that maternal and trophoblastic cells attaching to each other in placenta contain, as a rule glycogen. Glycogen cells in rat placenta and trophoblast cells of peripheral subpopulation of connective zone of placenta are similar in many respects. In this connection, a possible protective role of glycogen-containing cells, that probably favour the co-existence of maternal and embryo-derived cells in placenta, is discussed.

[Polyploidization dynamics of tertiary trophoblast giant cells in the rat placenta]. / Dinamika poliploidizatsii tretichnykh gigantskikh kletok trofoblasta v platsente krysy.

Polyploidization peculiarities of tertiary giant trophoblast cells during their active detaching from the ectoplacental cone and migrating into decidua basalis are investigated. On the 12th day of gestation, the ploidy of the majority of cell nuclei varies within 4-8c, although there are a few 16c and 32c nuclei. On the 13th and 14th days of gestation, the ploidy level of tertiary giant trophoblast cells enhances; 8c and 16c nuclei prevail, the percentage of 32c nuclei increases, 64c nuclei arising. The ploidy level of tertiary giant cell coincides with the average and/or maximum ploidy degree of precursor cell populations. The significance of polyploidy as indispensable condition of differentiation of the trophoblast cells that actively invade into maternal tissues is discussed.

[A quantitative study of Ag-positive nucleolar segments revealed by silver staining in the interphase nuclei of trophoblast cells from the rat placental connective zone]. / Kolichestvennoe izuchenie Ag-polozhitel'nykh uchastkov iadryshek, vyiavliaemykh v interfaznykh iadrakh kletok trofoblasta soedinitel'noi platsenty krysy metodom serebreniia.

A cytomorphological study was made of silver stained nucleoli in interphasic nuclei of trophoblast cells from the rat placenta connective zone, in addition to calculation of Ag-positive spherules in the nucleoli. The prevalent number of Ag-positive nucleolar spherules in the nuclei was 6, corresponding to the number of nucleolar organizers (NOR's) in the diploid chromosome complement of the rat. The mean number of Ag-positive spherules in the nucleoli progressively increase in the course of polyploidization from 2c to 32c; variability of the spherule number also increasing. The mean area of nucleoli is found to increase in proportion to the ploidy degree. A high correlation is found between the number of Ag-positive spherules and the area of nucleoli in the nucleus (r = 0.78). This appropriateness is exhibited at all the ploidy levels. The number of Ag-spherules and the area of nucleoli are found to depend slightly on the number of nucleoli. The possibility to use the number of Ag-positive spherules as a criterion of the activity of the NOR in interphasic nuclei is discussed.

[Ultrastructural features of the nucleus and cytoplasm of rat trophoblast cells of the connective zone of the placenta and labyrinth]. / Osobennosti ul'trastruktury iadra i tsitoplazmy kletok trofoblasta soedinitel'noi zony platsenty i labirinta krysy.

Ultrastructural organization of the rat trophoblast cells in the connective zone of placenta and labyrinth was investigated on the 12-14th days of gestation. A clear distinction was revealed in the cytoplasm ultrastructure of two cell subpopulations within the connective zone of placenta, i.e. glycogen and trophospongium cells. The former display a well defined network of long thin channels of granular endoplasmic reticulum situated mainly around the glycogen clusters. On the contrary, the latter are rich in the smooth endoplasmic reticulum but lacking glycogen accumulation. Differences in the nucleolar ultrastructure in these two cell subpopulations are not very considerable. A characteristic feature of glycogen cells is the presence of numerous round or oval small-fibrillar nucleolus-like bodies with the diameter of granules 20 nm. The trophoblast cells of the labyrinth are heavily laden with polysomes, which sometimes attach to short channels of the granular endoplasmic reticulum. Not often there occur short profiles of the agranular endoplasmic reticulum. Nucleolus-like bodies are found in all the cell types examined. This means that the nucleolus-like bodies may arise not only on the lampbrush chromosomes in the oocytes or polytene chromosomes, but also in the somatic cells which are capable of dividing mitotically.

[A comparative study of the main components of the nucleolus in different populations of rat trophoblast cells during differentiation. I. The nucleoli of the trophoblast cells in the labyrinth section of the placenta]. / Sravnitel'noe izuchenie osnovnykh komponentov iadryshka v raznykh populiatsiiakh kletok trofoblasta krysy v khode differentsirovki. I. Iadryshki kletok trofoblasta labirintnogo otdela platsenty.

The nucleolus undergoes some steps of structural transformation during differentiation of the labyrinth trophoblast cells. Primarily (on day 13 of gestation) the nucleolar components become rather disjoined. The nucleolus is composed of a loose net of strands of granulofibrillar and dense fibrillar components bearing fibrillar centers (FCs). Strands are separated by large lacunae. This rare-occurring type of nucleoli is replaced on the next (14th) day by the nucleolonemal type and later--by the compact nucleolar type. FCs with dense fibrillar component strands become extended into the masses of granulofibrillar component. Such transformations of nucleolar structure seem to be an expression of a fast-proceeding differentiation of the labyrinth trophoblast cells.

[A comparative study of the main components of the nucleolus in different populations of rat trophoblast cells during differentiation. II. The nucleoli of the trophospongium cells, the glycogen cells and the secondary giant cells]. / Sravnitel'noe izuchenie osnovnykh komponentov iadryshka v raznykh populiatsiiakh kletok trofoblasta krysy v khode differentsirovki. II. Iadryshki kletok trofospongiuma, glikogenovykh kletok i vtorichnykh gigantskikh kletok.

A comparative study was performed of the arrangement of different nucleolar components during differentiation of trophoblast cell populations in the junctional zone of placenta (glycogen cells and trophospongium) and in the secondary giant cells. Each cell type is characterized by specific interrelation of nucleolar components. Some glycogen cells show signs of segregation of nucleolar components: strands of nucleolar components with fibrillar centers (FCs) are displaced to the periphery of the nucleolus and contact with the perinucleolar chromatin. Large reticular nucleoli in trophospongium cells contain many FCs which are gathered into several "chains" by strands of dense fibrillar component. Such a "chain" has also been found in nucleoli of secondary giant cells, with greater number of FCs in each "chain". Relationship between the arrangement of nucleolar components and the level of cell differentiation is discussed.

[Changes in the relative arrangement of the chromosomes and nucleolus in developing mammalian oocytes during meiotic prophase I in relation to functional changes in the oocytes]. / Izmenenie vzaimnogo raspolozheniia khromosom i iadryshka v razvivaiushchikhsia ootsitakh mlekopitaiushchikh v khode profazy I meioza v sviazi s izmeneniem funktsional'nogo sostoianiia ootsitov.

Data on chromosome transformation in meiotic prophase I during mammalian oogenesis are summarized. The main peculiarity of the female meiosis in mammals is an unusually long diplotene stage which may be subdivided into four periods: 1) the early diplotene (up to the beginning of follicle formation); 2) the dictyotene or "diffuse diplotene", implying primordial follicle oocytes; 3) the most pronounced lampbrush chromosome stage coinciding with the large growth period; 4) the stage of chromosome inactivation and karyosphere formation corresponding to the terminal stage of oocyte development before ovulation. These stages are associated with changes in the transcriptional chromosome activity. A correlation is revealed between the spatial chromosome arrangement in the oocyte nucleus and the transcriptional activity. Some regularities are followed in the transformation of the main nucleolar component arrangement during meiotic prophase I in mammalian oocytes. At the late pachytene and at the early diplotene, a segregation of the main nucleolar components has been observed. These components are disposed in the direction: chromatin--fibrillar center--dense fibrillar component--granulo-fibrillar component. At the dictyotene, signs of nucleolar segregation are still observed. At the lampbrush chromosome stage, when the nucleus is most highly transcriptionally active, an integration of nucleolar components occurs. At the late diplotene--prediakinesis stage, i.e. in the course of transcriptional activity lowering and karyosphere formation, the secondary segregation of the main nucleolar components occurs. These move to the nucleolar periphery to be disposed around a large fibrillar mass which is gradually displacing the rest of the nucleolar components. The fibrillar mass formation in the preovulatory oocyte nucleoli is one of the peculiarities of the diplotene and prediakinetic mammalian oocytes.

[Polyteny and endomitosis in supergiant trophoblast cells of the gray vole Microtus subarvalis]. / Politeniia i éndomitoz v sverkhgigantskikh kletkakh trofoblasta seroi polevki Microtus subarvalis.

A cytomorphological study was made of peculiarly structured polytene chromosomes in supergiant trophoblast cells of Microtus subarvalis. The polyteny level was extremely high (over 1024C). The polytene chromosomes are characterized by a rather high degree of condensation of single chromosomes, and, as a consequence, close chromosome junctions and the typical disk pattern are lacking. The presence of complex nucleoli in the nuclei of these cells also testifies to a great detachment of chromonemes in polytene chromosomes of the studied supergiant trophoblast cells. Compared to other rodent species, a lower degree of chromoneme junction in the vole polytene chromosomes may cause their easy dissociation into single chromonemata, whose further condensation results in endomitotic chromosome formation. The chromosome depolytenization, earlier suggested from the analysis of interphase nucleus markers, has been traced here in detail. The process of polytene chromosome splitting was most obvious in the nucleolus-organizing chromosomes. A hony-combed nucleolus splits into numerous micronucleoli. The nucleus pattern becomes altered. Once in the polytene nucleus, chromosome bundles were located below the nuclear membrane and the central zone of the karyoplasm was not completely filled up. However, after dissociation of polytene chromosomes the whole karyoplasm was filled up with small nucleoli, and a thin layer of endomitotic chromosomes was seen beneath the nuclear membrane. The correlation between endomitosis and polyteny is discussed in terms of the dissociation of polytene chromosomes and formation of endomitotic chromosomes.

[Cells of the metrial gland: their relation to trophoblast cells and reproductive characteristics]. / Kletki metrial'noi zhelezy: ikh vzaimootnosheniia s kletkami trofoblasta i osobennosti reproduktsii.

Data on the origin, morphology and function of metrial gland cells are reviewed. Characteristic features of metrial gland cells are the availability of numerous eosinophilic granules lying near two round or oval nuclei and peripheral zone of the cytoplasm, generally devoid of organelles. This zone can generate pseudopodia-like projections. The notable peculiarity of metrial gland cells involves their ability to penetrate into blood vessels, to migrate towards the embryo, and to achieve the ectoplacental cone. The majority of metrial gland cells is accumulated in the decidua basalis zone where the tertiary trophoblast cells usually migrate. The metrial gland cells seem to constitute a cell population analogous to that of decidual cells. Data on the protective role of metrial gland cells are discussed. The metrial gland cells are proven to be polyploid. Polyploid nuclei are found both in mononucleate and binucleate cells. Acytokinetic mitosis is presumably a way leading to polyploidization of metrial gland cells.

[Characteristics of trophoblast cell reproduction in the connective zone of the rat placenta. I. Determination of the degree of ploidy and the number of Barr bodies in interphase nuclei]. / Osobennosti reproduktsii kletok trofoblasta soedinitel'noi zony platsenty krysy. I. Opredelenie stepeni ploidnosti i chisla telets Barra v interfaznykh radrakh.

The degree of ploidy in the interphase nuclei was determined in the connective zone of the rat's placenta on days 13 and 14 of embryo development. On day 13, the ploidy in the majority of nuclei was 2c or 4c; on day 14, the 4c nuclei were dominating, the share of 8c nuclei increasing. The number of Barr's bodies in each nucleus of the placental connective zone tends to increase with the increase in ploidy level. This is an evidence of a "genuine polyploidy" as a mechanism of the initial polyploidization of the given cell population.

[Characteristics of trophoblast cell reproduction in the connective zone of the rat placenta. II. Determination of the degree of ploidy of mitotic shapes]. / Osobennosti reproduktsii kletok trofoblasta soedinitel'noi zony platsenty krysy. II. Opredelenie stepeni ploidnosti mitoticheskikh figur.

Morphological and cytophotometric studies have been made on polyploidization of placenta connective zone cells. Measurement of the DNA content in mitotic figures show that within a period of development ranging from day 13 to day 14 the bulk of mitoses (up to 25%) become tetraploid and octaploid. This may suggest that polyploidization of placenta connective zone cells proceeds via incomplete polyploidizing mitoses. Among tetraploid and octaploid mitotic figures, there are those corresponding to all the mitotic stages, from prophase to telophase. Consequently, mitosis in tetraploid and octaploid cells can reach telophase. In such cases polyploidization is likely to follow the acytokinetic mitotic pattern. A question of a certain maximum level of polyploidy that may be reached by cells due to the incomplete mitosis is discussed.

[DNA content of the nuclei of secondary giant cells of the rat trophoblast at different phases of the polytene nucleus cycle]. / Izuchenie soderzhaniia DNK v iadrakh vtorichnykh gigantskikh kletok trofoblasta krysy na raznykh fazakh tsikla politennogo iadra.

A cytophotometric study of DNA content has been made for secondary trophoblastic giant cells, which differ morphologically in relation to the stage of the cycle of the polytene nucleus. The ploidy rate varying from 16c to 512c. It is shown that the DNA content of the nuclei with polytene chromosomes in phase G is more stable, corresponding to the 2c multiple DNA content. Unlike, reticular nuclei in phase S do not present clear-cut peaks on a histogram of DNA. Ratios of nuclei with unequal ploidy differ depending on the structure of these nuclei.

Genome multiplication of extravillous trophoblast cells in human placenta in the course of differentiation and invasion into endometrium and myometrium. II. Mechanisms of polyploidization.

Peculiarities of the structure of interphase nuclei, mitotic activity, and Ki-67 protein intranuclear immunolocalization were studied to elucidate mechanisms of genome multiplication in proliferative and differentiating invasive extravillous trophoblast cells in the human placenta. The presence of numerous chromocenters was shown to be a characteristic feature of proliferative cell nuclei of both villous and extravillous trophoblast. At the beginning of extravillous trophoblast cell differentiation, i.e. in the proximal part of cell columns, some amount of cells with large nuclei containing enlarged chromocenters were found. DNA content was measured simultaneously with counting the number of chromocenters in similarly looking nuclei of squash preparations of placental villi. The increase in the ploidy level up to 4c-8c, accompanied by a slight increase in the number of chromocenters being not proportional to the ploidy level and not exceeding the diploid number of chromosomes of the human genome, was demonstrated. This suggests that genome multiplication of extravillous trophoblast cells may be accomplished by endoreduplication. In addition, pictures of endomitosis were seen at early steps of differentiation of EVT cells. The lack of polyploid mitotic figures or any obvious polyploidizing or restitutional mitoses suggests that these are not of considerable importance in genome multiplication of human EVT cells. However, the prevalence of metaphases at the boundary of the distal part of cell columns suggests that restitutional mitoses may be involved, even partly, in human trophoblast cell polyploidization. At later steps of differentiation, i.e. in the distal part of cell columns, the nuclear structure obviously changes, with a uniform "network" chromatin arrangement prevailing, whereas numerous chromocenters and features of endomitosis are no longer seen. The pattern of Ki-67 protein immunolocalization is also changing along the invasive pathway. In the proliferating stem cells and trophoblast cells of the proximal part of cell columns, Ki-67 was localized in the karyoplasm, chromocenters and numerous small nucleoli, whereas in the distal part of cell columns this protein was detected predominantly in 1-2 large nucleoli. The comparative analysis of the literature data on Ki-67 localization at different stages of cell cycle provided another evidence that EVT cells in the course of invasion may switch to the endoreduplication cycle. In agreement with the relevant report on rodent placentation, our present data suggest that acquirement of an invasive phenotype of EVT cells is accompanied by switching from mitotic division to endoreduplication cycle.

Genome multiplication of extravillous trophoblast cells in human placenta in the course of differentiation and invasion into endometrium and myometrium. I. Dynamics of polyploidization.

Polyploidization of the extravillous trophoblast (EVT) cells at different stages of differentiation and invasion into the uterine wall in human placenta has been studied. An increase in the ploidy level of EVT cells in the course of their differentiation within cell columns (CC) was shown. Stem cells were mainly diploid (86.2%); incidence of polyploid nuclei of highly proliferative cells of the proximal part of CC increased progressively. In the distal part of CC, where EVT cells did not divide mitotically, polyploid cells prevailed, with 58.0 and 3.5% nuclei being 4c and 8c, respectively. The highest percentage of polyploid cells was found in the population of EVT cells attached directly to the surface of the decidualized endometrium: percentage of tetraploid cells turned out to be 74.7% and the share of octaploid nuclei rose up to 4.9%; however, there appeared a few (0.3%) 16c cells. The majority of EVT cells invading the decidualized endometrium were polyploid, the share of octaploid and hexadecaploid cells rose up to 9.7 and 1.4%, respectively. On the other hand, the percentage of diploid cells also increased up to 29.2% as compared to EVT cells attached to decidua (20.0%). The same tendency proved to be even stronger in myometrium: the share of diploid EVT cells increased up to 46.0%, a prominent amount of tetraploid (45.1%) and highly polyploid (8c and 16c) cells retained in the EVT cell population (7.4 and 1.1%, respectively). Immunohistochemical staining of Ki-67 protein (MIB1), which labels cells held in the cell cycle, showed a high incidence of MIB1-positive stem cells (93.7%) and the EVT cells of the proximal part of CC (85.5%) characterized by high mitotic activity. A lower MIB1-positivity (43.2%) was found in the distal part of CC, whereas invasive EVT cells showed no MIB1-labeling. The presence of MIB1-positive nuclei in the distal part of CCs in the absence of mitoses, taken together with data on polyploidization of these cells, indicates their switch to the endoreduplication cycle. As a whole, the data obtained evidence that differentiation of EVT cells of the invasive pathway is accompanied by polyploidization. However, in a population of trophoblast cells capable of most profound invasion (up to myometrium), the proportion of diploid cells rose. These results suggest that the human cytotrophoblast invasion into the uterine wall requires an optimum, not the highest, ploidy level, whereas highly polyploid cells may form a subpopulation at the border between the maternal and fetal parts of placenta.

[Quantitative investigation of reproduction of condensed chromatin of sex chromosomes during trophoblast cell polyploidization and endoreduplication in the East European field vole Microtus rossiaemeridionalis]. / Kolichestvennoe issledovanie reproduktsii kondensirovannogo khromatina polovykh khromosom pri poliploidizatsii i éndoreduplikatsii kletok trofoblasta vostochonoevropeiskoi polevki Microtus rossiaemeridionalis.

Simultaneous measurement of DNA content in cell nuclei and condensed chromatin bodies formed by heterochromatized regions of sex chromosomes (gonosomal chromatin bodies, GCB) has been performed in two trophoblast cell populations of the East-european field vole Microtus rossiaemeridionalis, namely in the proliferative population of trophoblast cells of the junctional zone of placenta and in the secondary giant trophoblast cells. One or two gonosomal chromatin bodies have been observed in trophoblast cell nuclei of all embryos studied (perhaps both male and female), In the proliferative trophoblast cell population, characterized by low ploidy levels (2c-16c), and in the highly polyploid population of secondary giant trophoblast cells (16c-256c), the total DNA content in GCB increased proportionally to the ploidy level. In separate bodies, the DNA content rose also in direct proportion with the ploidy level seen in the nuclei with both one and two GCBs in the two trophoblast cell populations. A certain increase in percentage of the nuclei with 2-3 GCBs was shown in the nuclei of the junctional zone of placenta; this may be accounted for by genome multiplication via uncompleted mitoses. In the secondary giant trophoblast cell nuclei (16c-256c), the number of GCBs did not exceed 2, and the share of nuclei with two GCBs did not increase, thus suggesting the polytene nature of sex chromosome in these cells. At different poloidy levels, the ratio of DNA content in the nucleus to the total DNA content in GCB did not change significantly giving evidence of a regular replication of sex chromosomes in each cycle of genome reproduction. In all classes of ploidy, the mean total DNA content in trophoblast cell nuclei with single heterochromatic body was less than in the nuclei with two and more GCBs. This may indicate that a single GCB in many cases does not derive from the fusion of two GCBs. To put it another way, in the nuclei with one GCB and in those with two or more GCBs, different chromosome regions may undergo heterochromatization. The regularities observed here are, most probably, associated with the peculiarities in the structure of X- and Y-chromosomes in a range of species of Microtus (M. agrestis, M. rossiaemeridionalis, M. transcaspicus). As a result, gonosomal chromatin bodies may include large blocks of both constitutive heterochromatin of X- and Y-chromosomes (in male and female embryos) and inactivated euchromatin of "lyonized" X-chromosome in female embryos. Therefore the presence of two or more GCBs in trophoblast cells of M. rossiaemeridionalis may be accounted for by both polyploidy and functional state of the nucleus, in which gonosomal constitutive heterochromatin and inactivated euchromatin form two large chromocenters rather than one. The differences in DNA content in GCBs in the nuclei with one and two GCBs seem to be an indirect indication that the two chromocenters may be formed by two different gonosomes, with the extent of their heterochromatization being higher than that in the nuclei with one GCB. GCBs in the trophoblast cells of M. rossiaemeridionalis are observed not only at the early developmental stages, as it was observed in rat at the first half of pregnancy (Zybina and Mosjan, 1967), but also at the later stages, up to the 17th day of gestation. At these stages, the nuclei with non-classical polytene chromosomes rearrange to those with a great number of endochromosomes, probably because of disintegration of chromosomes into oligotene fibrils. However, it does not seem unlikely that this process may involve heterochromatized gonosomal bodies, since only one or two large GCBs can be seen in the nuclei as before. The presence of prominent blocks of constitutive heterochromatin seems to favor a closer association of sister chromatids in polytene chromosomes, which prevents their dissociation into endochromosomes with the result that polyteny of sex chromosomes in the field vole trophoblast is probably retained during a longer period of embryonic development.

[Polyploid mitoses in mink trophoblast cells]. / Poliploidnye mitozy v kletkakh trofoblasta norki.

Mitotic figures in the mink placental trophoblasts have been observed under the light microscope using actions and air-dried preparations. The tetra- and octaploid metaphase chromosome spreads were found on Giemsa-stained air-dried preparations. A high percentage (up to 80%) of abnormal metaphases, including k-mitoses as well as a portion of restitution anaphases, was revealed on sections of the placental trophoblast suggesting a possible block of mitosis at the meta- and anaphase. Therefore, it is very likely that genome multiplication in a portion of placental trophoblast cells in mink involves block of mitoses at meta- and anaphase followed by restitution. The chromosomal arrangement on metaphase spreads in part of cells showed a degree of separation of the whole di- and tetraploid chromosome sets within tetraploid and octaploid chromosome plates. Several spreads exhibited some allocycly of diploid chromosome sets inside the polyploid metaphases. It is not inconceivable that such an arrangement may reflect some autonomy of low-ploidy chromosome sets within the polyploid trophoblast cells in mink.

[Characteristics of nuclear structure and karyosphere formation in mink oogenesis]. / Osobennosti struktury iadra i formirovaniia kariosfery v oogeneze norki.

Changes in nuclear structures have been followed in oocytes of adult minks, from the primordium follicle stage to the Graaf vesicle stage. The followed dynamics well compared with what has been known for other mammals studied. With the mink oogenesis, an intensive production of nucleolus-like bodies occurs: these may be as many as 100 per nucleus. On later stages of occyte growth, ring-like extranuclear bodies were found which have not been observed for other mammals. With the mink, karyosphere formation is generally similar to that in other mammalian species examined, being, however, observed a bit earlier than usually, i.e. on the stage of multilayered follicle with the antrum. Due to this fact, the karyosphere, with the mink, persists longer. With the mink, contrary to other species examined, the formation of the karyosphere as a dense Feulgen-positive body is not accompanied with a total disappearance of the nuclear envelope and nucleolus-like bodies.