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PubMed1980

[Preliminary study of stages of human meiosis in spermatocytes and ovocytes (author's transl)].

Human meiotic chromosomes, from spermatocytes and ovocytes, are described after observations of whole mount preparations under E.M. Small testicular and ovarian fragments are put in distillated water, then macerated; the cell suspension is spread on the surface of sheet copper grids covered with formvar plus collodion films. After dehydratation interesting stages are selected under L.M. before observations under E.M. Zygotene and pachytene are the most common stages. During pachytene the chromomeres are well individualized; the synaptonemal complex may be observed; chromatin fibers connect the chromosomes to nuclear pores, interchromosomal fibers joint the bivalents. Zygotene and pachytene bivalents are very similar in the male and the feminine germ cells.

Chromatin↗

PubMed1986

The behavior and morphology of the X and Y chromosomes during prophase I in the Sitka deer mouse (Peromyscus sitkensis).

Surface-spread, silver-stained primary spermatocytes from individuals of the Sitka deer mouse (Peromyscus sitkensis) were analyzed by electron microscopy. Pairing of the X and Y chromosomes is initiated at early pachynema and is complete by mid pachynema. The pattern of sex chromosome pairing is unique in that it is initiated at an interstitial position, with subsequent synapsis proceeding in a unidirectional fashion towards the telomeres of the homologous segments. One-third the length of the X and two-thirds the length of the Y are involved in the synaptonemal complex of the sex bivalent. Various morphological complexities develop in the heteropycnotic (unpaired) segments as pachynema progresses, but desynapsis is not initiated until diplonema. Analysis of C-banded diakinetic nuclei indicated that sex chromosome pairing involves the heterochromatic short arm of the X and the long arm of the heterochromatic Y. An interstitial chiasma between the X and Y was observed in the majority of the diakinetic nuclei. The observation of a substantial pairing region and chiasma formation between the sex chromosomes of these deer mice is interpreted as indicating homology between the short arm of the X and the long arm of the Y.

Animals↗

PubMed1983

Cytologic evidence for three human X-chromosomal segments escaping inactivation.

Early replication of prometaphasic human sex chromosomes was studied with the bromodeoxyuridine (BrdU)-replication technique. The studies reveal that two distal segments of Xp, including bands Xp 22.13 and Xp 22.3, replicate early in S-phase and therefore may not be subject to random inactivation. Furthermore, the replication of these distal segments of Xp occurs synchronously with those of the short arm of the Y chromosome including bands Yp 11.2 and Yp 11.32. These segments of Xp and Yp correspond well to the pairing segment of the X and Y chromosomes where a synaptonemal complex forms at early pachytene of human spermatogenesis. The homologous early replication of Yp and the distal portion of Xp may be interpreted as a remnant left untouched by the differentiation of heteromorphic sex chromosomes from originally homomorphic autosomes. A third early replicating segment is situated on the long arm of the X chromosome and corresponds to band Xq 13.1. This segment may be correlated with the X-inactivation center postulated by Therman et al. (1979).

Bromodeoxyuridine↗

PubMed1993

Expression of the Saccharomyces cerevisiae RAD50 gene during meiosis: steady-state transcript levels rise and fall while steady-state protein levels remain constant.

In Saccharomyces cerevisiae, the RAD50 gene is required for repair of X-ray and MMS-induced DNA damage during vegetative growth, and for synaptonemal complex formation and genetic recombination during meiosis. We show below that the RAD50 gene encodes major and minor transcripts of 4.2 and 4.6 kb in length which differ primarily at their 5' ends. Steady-state levels of both RAD50 transcripts increase coordinately during meiosis, reaching maximal levels midway through meiotic prophase, about 3 or 4 h after transfer of cells to sporulation medium. The 5' ends of the major RAD50 transcript in both meiotic and vegetative cells map to the same cluster of sites approximately 20 bp upstream of the amino-terminal ATG of the RAD50 coding sequence. We conclude that the increased RAD50 transcript level observed during meiosis does not reflect utilization of a new promoter. In contrast, steady-state levels of Rad50 protein do not increase during meiosis. Thus, changes in RAD50 transcript levels are not necessarily accompanied by commensurate changes in Rad50 protein levels. Possible explanations are considered.

Base Sequence↗

PubMed1980

Ultrastructural characterization of the sex chromosomes during spermatogenesis of spiders having holocentric chromosomes and a long diffuse stage.

An ultrastructural study has been made of spermatogenesis in two species of primitive spiders having holocentric chromosomes (Dysdera crocata, male X0 and Sergestria florentia X1X2O). Analysis of the meiotic prophase shows a scarcity or absence of typical leptotene to pachytene stages. Only in D. crocata have synaptonemal complex (SC) remnants been seen, and these occurred in nuclei with an extreme chromatin decondensation. In both species typical early prophase stages have been replaced by nuclei lacking SC and with their chromatin almost completely decondensed, constituting a long and well-defined diffuse stage. Only nucleoli and the condensed sex chromosomes can be identified. - In S. florentina paired non-homologous sex chromosomes lack a junction lamina and thus clearly differ from the sex chromosomes of more evolved spiders with an X1X20 male sex determination mechanism. In the same species, sex chromosomes can be recognized during metaphase I due to their special structural details, while in D. crocata the X chromosome is not distinguishable from the autosomes at this stage. - The diffuse stage and particularly the structural characteristics of the sex chromosomes during meiotic prophase are reviewed and discussed in relation to the meiotic process in other arachnid goups.

Animals↗

PubMed1982

Dense bodies in silver-stained spermatocytes of the Chinese hamster: behavior and cytochemical nature.

From the silver staining behavior of various organelles in the nucleus we have divided meiotic prophase (leptotene to the diffuse stage) of the male Chinese hamster into five stages. Components within the nucleus, such as synaptonemal complex (SC), sex bivalent (SB), nucleolus organizer regions (NORs), chromatin and the dense bodies, showed a characteristic feature in each stage of meiotic prophase. The lampbrush chromosome stage was found to be followed by the diffuse stage. The chromatin around SC began to be organized at early pachytene and formed a brush-like structure at late pachytene. During early prophase stages a dramatic change in SB morphology occurred. Three types of morphology of SB were recognized: (1) the XY pair with long synapsis and fusiform or diffuse thickening of the unpaired portions (late zygotene and early pachytene), (2) desynapsed, branched, and anastomosed axes seen at late pachytene. Two types of the dense body were found during meiotic prophase; the double body in early stage (leptotene to early pachytene) and the single body in later stages (mid pachytene to diffuse stage). The small precursors of the double body existed at early leptotene but they increased in size and also changed the silver stainability during zygotene, becoming the characteristic double body consisted of one light body (L-body) and one dark body (D-body). These two bodies can also be recognized after Giemsa or acridine orange (AO) staining. The L-body fluoresced reddish orange after AO staining. The single body, which is probably formed by amalgamation of the D- and the L-bodies, showed a staining reaction similar to that of the D-body. Data from pancreatic lipase and protease treatments suggest that the D-body contained a lipoprotein.

Animals↗

PubMed1989

Anti-topoisomerase II recognizes meiotic chromosome cores.

At meiotic prophase the chromatin becomes arranged in loops on newly formed chromosome cores. The cores of homologous chromosomes become aligned in parallel and thus form the synaptonemal complex (SC), a structure found in the meiocytes of nearly all recombinationally competent, sexually reproducing organisms. We report that two polyclonal antibodies against topoisomerase II (topo II), which recognize the mitotic metaphase chromosome scaffold give, at pachytene, a positive immunocytological reaction with the chromatin and, predominantly, with the cores and centromeric regions of the paired chromosomes. It therefore appears that during meiotic prophase, topo II - a DNA-binding enzyme implicated in transient double-strand breaks, chromosome condensation, and anaphase separation - is associated with the chromatin and SCs of the pachytene and diplotene chromosomes.

Animals↗

PubMed1984

Pachytene chromosomes in male and female mice heterozygous for the Is(7;1)40H insertion.

Pairing of pachytene chromosomes was studied in oocytes and spermatocytes of mice heterozygous for the male-sterile Is(7;1)40H insertion using light and electron microscopy for synaptonemal complex analysis in surface-spread, silver-stained preparations. The data comprised four males and four female embryos. The insertion/deletion configurations appeared as either two bivalents or one quadrivalent in both sexes, but the proportion of bivalents was higher in oocytes. Some insertion and deletion bivalents showed synaptic adjustment. The insertion/deletion configurations were associated with, or adjacent to, the XY bivalent in the majority of spermatocytes. End-to-end association of different bivalents was more frequent in oocytes than in spermatocytes. It is suggested that physiological differences between male and female gametocytes may lead to the difference in their reproductive potential.

Animals↗

PubMed1986

The mechanism of autosomal synapsis and the substaging of zygonema and pachynema from deer mouse spermatocytes.

Surface-spread and silver-stained spermatocytes of Peromyscus maniculatus and P. sitkensis were analyzed in order to develop criteria for the recognition of meiotic substages from early zygonema through early diplonema. The continuous sequence of changes in the morphology of the autosomal axes (lateral elements) of the synaptonemal complexes (SC), sex chromosome axes, and nucleoli enabled the recognition of three substages of zygonema and five of pachynema. The proposed system of subdivision is compatible with descriptions of comparable data from Chinese hamsters and laboratory mice with differences being primarily associated with the timing of sex chromosome synapsis and desynapsis. Within the substages, cytogenetically important details of the synaptic mechanism in deer mice were noted. Autosomal synaptic initiation in deer mice is apparently uniterminal, involving the distal (noncentromeric) end of the homologs. Subsequent pairing is unidirectional towards the centromeric end. Additionally, during mid and late zygonema the homologous axes of late pairing regions of some autosomes were characterized by substantial length differences. These lateral element length differences were not maintained into pachynema and it is hypothesized that differences in the amount of material in the heterochromatic short arms of these species may be subject to synaptic adjustment.

Animals↗

PubMed1986

Heterosynapsis and suppression of chiasmata within heterozygous pericentric inversions of the Sitka deer mouse.

The patterns of chromosomal pairing and chiasma distribution were analyzed in male Sitka deer mice (Peromyscus sitkensis) polymorphic for terminally positioned pericentric inversions of chromosomes 6 and 7. G- and C-banding of somatic metaphases indicated that the inversions involved 30% and 40% of chromosomes 6 and 7, respectively. Analysis of silver-stained synaptonemal complexes in surface-spread zygotene and pachytene nuclei from heterozygous individuals revealed that inversion loops were not formed. The inverted segments proceeded directly to heterosynapsis without an intervening homosynaptic phase, and the heteromorphic bivalents remained straight-paired throughout pachynema. C-banded pachytene nuclei corroborated the occurrence of heterosynapsis, as the heteromorphic bivalents exhibited nonaligned centromeres. Analysis of diplonema and diakinesis indicated that crossing over had not occurred within the heterosynapsed inverted segments. The observation of chiasma suppression within the inversions indicates that pericentric inversion heterozygosity does not lead to the production of unbalanced gametes. Heterosynapsis of the inverted segments during zygonema and pachynema and the resulting chiasma suppression therefore represent a meiotic mechanism for the maintenance of pericentric inversion polymorphisms in this population of P. sitkensis.

Animals↗

PubMed1976

A cytological study of micronuclear elongation during conjugation in Tetrahymena.

Micronuclear elongation is the first major event in a series of nuclear changes occurring during the sexual stage of the life cycle of Tetrahymena. Beginning at about one hour after cells of complementary mating types have conjugated, the micronucleus leaves its recess in the macronucleus and swells slightly. This is accompanied by a reorganization of its chromatin from a reticular to a solid body. In the next stage the micronucleus assumes an egg shape, a development concomitant with the appearance of microtubules. While the chromatin "spins out" from the dense body, and microtubules increase in number, the nucleus assumes a spindle shape. During the elongation, which increases the length of the nucleus some fifty fold, microtubules are prominent in clusters just internal to the nuclear membrane, and parallel to the longitudinal axis of the nucleus. When elongation is completed the nucleus is curved around the macronucleus. Internally, partially condensed strands of chromatin are located off-center, towards the macronuclear side, and the density of the microtubules is diminished. At all the stages, DNA is located throughout the nucleus; neither discrete chromosomes nor synaptonemal complexes are seen. Occasionally cytoplasmic membrane systems are seen fused to the nuclear envelope which retains the typical appearance of a double membrane with pores.

Animals↗

PubMed1979

Light microscope analysis of meiotic prophase chromosomes by silver staining.

A method is described for the silver staining of the synaptonemal complex in surface-spread mammalian spermatocytes for light microscope examination. The method is quick, reliable, of broad applicability, and provides a means of making karyotype analysis at meiotic prophase. Many hundreds of suitable cells can be examined in an average preparation in a relatively short space of time. It has so far been applied only to mammalian spermatocytes, but could be used for karyotype analysis in oocytes of mammals and also applied to gonocytes of non-mammalian species.

Animals↗

PubMed1985

Silver staining of meiotic chromosomes in the fungus, Coprinus cinereus.

We have taken advantage of the synchronous meiotic process in the basidiomycete Coprinus cinereus to develop a simple and rapid method to selectively stain meiotic chromosomes and nucleoli in this fungus without prior removal of the cell wall. Electron microscopic examination of these silver-stained chromosomes indicated that the lateral elements of the synaptonemal complexes were prominently stained, and terminal attachment plaques were apparent. We found that a translocation quadrivalent could be recognized easily in the light microscope using these methods. The procedures appear suitable for the characterization of chromosome rearrangements in this small genome, and should facilitate cytogenetic analysis in this fungus.

Basidiomycota↗

PubMed1982

Homologous early replication patterns of the distal short arms of prometaphasic X and Y chromosomes.

Replication studies on prometaphasic human sex chromosomes reveal a distinct early replicating segment on both distal Xp ad Yp. These segments correspond to high resolution bands Xp22.3 and Yp11.3. The findings demonstrate synchronous replication of these parts of the sex chromosomes and correspond to the comparatively long stretches of Xp and Yp that participate in a synaptonemal complex. Furthermore these observations are compatible with Polani's view that suggests homologous segments with similar genetic information on both sex chromosomes (Polani 1982).

Chromosome Banding↗

PubMed1989

Satellite DNA I in chromatin loops of rat pachytene chromosomes and in spermatids.

Biotinylated rat satellite DNA I probe p93-50 was used to visualize the chromatin of surface-spread rat pachytene chromosomes. Fluorescein isothiocyanate (FITC)-conjugated avidin produces a beaded fluorescence pattern along the chromatin loops that insert in the centromeric region of the synaptonemal complex (SC), the paired cores of homologous chromosomes. The number of fluorescent beads ranges from zero for centromeres without satellite DNA I homologous to probe p93-50, to several hundred for satellite-rich centromeric regions. For the chromosomes that can be identified, the relative amount of satellite DNA is chromosome specific. No satellite DNA I was detected at the non-centromeric ends of the chromosomes or interstitially. DNase-digested nuclei or isolated SCs did not have detectable amounts of satellite DNA in the centromeric regions of the chromosomes or in the residual SCs. The fate of the satellite DNA was followed during spermiogenesis. In the round spermatid the centromeric regions, which appear to be attached to the nuclear envelope, are still distinct and have converging loops of fluorescent chromatin. At later stages there are fewer but still bright fluorescent patches. Satellite DNA I is still detectable in the mature sperm head. These results demonstrate the organization of satellite DNA I in the chromatin loops at the centromeric regions, and they forecast the analysis of chromosome organization in unprecedented detail with a variety of probes in surface spreads of meiotic prophase chromosomes.

Animals↗

PubMed1989

Transmission of three radiation-induced translations in the Syrian hamster. I. Chromosome studies of male meiosis: pachytene to metaphase II.

Chromosome behaviour during meiosis in male Syrian hamsters heterozygous for one of three translocations was analysed as part of a study of the transmission of these structural changes. Synapsis was studied using preparations of synaptonemal complexes, and chiasmate associations and the results of anaphase I segregation were studied in air-dried preparations of metaphases I and II respectively. The main findings were: (i) that, at least in the two trivalent-forming translocations, there is no simple relationship between either the frequency or the extent of synapsis and chiasma formation between the chromosomes involved in the translocation; (ii) that the presence of a univalent in a substantial proportion of metaphase I cells does not necessarily lead to irregular segregation as judged by analysis of metaphase IIs; and (iii) conversely, that in translocation heterozygotes in which metaphase I contains the chromosomes involved in the translocation as a quadrivalent or as two bivalents, with no univalents or trivalents, unexpected numerical segregation can be found. The observations of meiotic chromosomes behaviour reported here show that it is not always possible to predict the effects of structural change, or to determine the basis of these effects, from an analysis of any stage of meiosis taken in isolation, or from an analysis of an apparently similar change.

Animals↗

PubMed1981

Chromosome behaviour during early meiotic prophase of mouse primary spermatocytes.

A correlative light- and electron microscopical study has yielded both a description of chromosome behaviour during early meiotic prophase in mouse spermatocytes, and an accurate timing of the zygotene and early pachytene stage. The light microscopic part of the study consists of an analysis of orcein stained or C-banded, air-dried preparations, and series of separately squashed sections of seminiferous tubules. The mice used for air-dried preparations were treated with hyroxyurea and triaziquone, which reduces the spermatocyte population to a small, well-defined group of cells in meiotic interphase. The development of the restricted spermatocyte population is followed during four days. The electron microscopical study consists of an investigation of synaptonemal complexes in thin sections of seminiferous tubules in various stages of the epithelial cycle. It turned out that synaptic pairing starts about half a day after the end of the DNA replication phase. At most one and a half day later chromosome pairing is completed. In very early pachytene spermatocytes a sex vesicle is not visible. The orientation of the X and Y chromosome into a sex vesicle starts about one day after the beginning of the pachytene stage. The existence of a leptotene stage is discussed. The presence of a pre-leptotene contraction phase in male mice is excluded.

Animals↗

PubMed1978

"Nodules" in the achiasmatic meiosis of Bithynia (Mollusca, Prosobranchia).

In spermatocytes of Bithynia leachi and B. tentaculata, synaptonemal complexes with nodules in an achiasmatic meiosis were demonstrated for the first time by means of the spreading technique. The number and position of the nodules are different in each species and are independent of the length of the chromosomes. Although no crossing over occurs in Bithynia chiasma-like terminal connections of the chromosomes are preserved in diakinesis by the presence of the nodules. --Enzymatic treatments revealed information regarding their composition of alkaline proteins, RNA and DNA. In all proteolytic digestions the nodules more of less retain their position in the central region, while they disappear immediately if SCs are exposed to DNase. On the basis of the results a model of the composition and the function of nodules can be developed considering the concept of a lampbrush-organization of the chromosomes in early meiotic prophase.

Animals↗