Synaptonemal complex karyotyping in spermatocytes of the Chinese hamster (Cricetulus griseus)

Relative length is a constant and distinctive characteristic for each autosomal SC, despite variations in absolute length from cell to cell. Arm ratio is distinctive for each SC except for two of the three sub-acrocentrics, and serves, together with relative length, for identification. The constancy of relative length and arm ratios indicates biological stability and lack of physical distortion in these spread preparations. There is a 11 relationship between relative lengths of autosomal SCs and mitotic autosomes; their arm ratios are also similar. These close parallels provide strikingly similar SC and somatic karyotypes. Variability was observed in sub-acrocentric arm ratios and in lengths of unpaired X and Y axes, correlated with the presence of constitutive heterochromatin. — Utilizing progressive differentiations of the X and Y chromosomes for staging, it is demonstrated that autosomal SCs decrease in length from late zygotene to mid-pachytene, and then increase at late pachytene. Within a nucleus, synchrony of length changes is maintained. It is concluded that the factors governing autosomal SC length are regular for any given bivalent from cell to cell, and may be related to those that control somatic autosome length relationships. — The X and Y axes differ quantitatively as well as qualitatively from autosomal SCs. The SC portion of the X and Y is constant in length through most of pachytene; the unpaired axes shorten and lengthen, but not in proportion to autosomal SCs. X and Y relative lengths and arm ratios vary throughout pachytene and do not maintain proportionality with somatic values. The evidence suggests, but does not prove, that the long arm of the X is paired with the short arm of the Y. — Twists occur in autosomal SCs at increasing frequencies throughout pachytene but cannot account for length changes. The number of twists per SC is directly proportional to SC length. Intertwining of SCs is random and proportional to SC length. End-to-end associations of autosomal SCs appear to be random; however, the ends of the X and Y are less often involved in such connections. — The length of axial material in all chromosomes at pachytene, expressed as an equivalent length of DNA double helix, represents 0.013% of the diploid DNA complement.     

Silver-Stained accessory structures on human sex chromosomes

Summary Using a combination of silver-staining and light microscopic techniques on human male meiotic preparations, it is feasible to study the morphology and behavior of both autosomal synaptonemal complexes and sex chromosome axes. During leptotene and early zygotene, the X and Y chromosomes are separate; their axes appearing as thin, filamentous structures. During late zygotene/early pachytene, the sex chromosomes come close to each other and a distinct sex vesicle is formed. We confirm the existence of a short synaptonemal complex between the terminal ends of the X and Y chromosomes. In our preparations, a number of accessory structures can be seen along the axes of the sex chromosomes. These structures appear to be similar in morphology to those previously observed in several other mammalian species.     

Chromosomes and DNA of Mus

Using C-banded preparations of Mus dunni it is possible to study the behavior of constitutive heterochromatin in early stages of meiotic prophase. The X and the Y chromosomes, both of which contain a large amount of heterochromatin, lie apart in leptotene but move toward each other during zygotene. They then form the sex vesicle at late zygotene. In autosomes zygotene pairing appears to start from the telomeric ends. The centromere of the Y chromosome associates end-to-end with the terminal end of the long arm of the X chromosome. The autosomal heterochromatic short arms show forked morphology in certain bivalents at pachytene, suggesting probable incomplete synapsis.     

Heterochromatin, synaptonemal complex, and NOR activity in the somatic and germ cells of a male domestic dog, Canis familiaris (Mammalia, Canidae)

C-banding and silver staining of the somatic and germ cells of the male domestic dog. Canis familiaris, have shown that: (1) the amount of C-banding is small compared to most other mammalian species, (2) three pairs of autosomes have nucleolus organizer regions (NORs) at the terminal ends of their long arms, whereas the Y chromosome has an NOR on the terminal end of the short arm, (3) the organization of the synaptonemal complex (SC) is similar to that of other mammalian species, (4) a distinct SC is formed between the long arm of the Y chromosome and probably the short arm of the X chromosome, and (5) the differential axes of both sex chromosomes do not demonstrate fusiform thickenings nor do they stain darkly with silver as do the XY bivalents in many other mammalian species.     

Electron microscopic observations on the synaptonemal complex of spermatocytes of the Giant Panda (Ailuropoda melanoleuca)

Surface-spread and silver-stained preparations of spermatocytes from a giant panda were observed by electron microscopy for synaptonemal complex karyotyping. Ten pachytene spermatocyte nuclei were selected for length quantitation of SC. The mean relative lengths and centromeric indices of each SC agreed closely with those of the mitotic chromosomes. The pairing between lateral elements of autosomal chromosomes starts at early zygotene and leads progressively along their length to complete pairing at pachytene. The whole Y is paired with 1/3 length of X at mid-pachytene. The morphology of X and Y chromosome axes and the nonhomologous pairing of X and Y is discussed.     

Meiotic behavior of gonosomically variant females of Akodon azarae (Rodentia, Cricetidae)

The meiotic behavior of sex chromosomes has been investigated in variant females of Akodon azarae, both in pachytene oocytes and metaphase I. In somatic cells, these females have a heteromorphic sex pair, in which the minor chromosome has been previously interpreted as a major deletion of the long arm of the X chromosome (dX). After microspreading for synaptonemal complex analysis, pachytene oocytes show two axes of very different lengths (100:17.1), which correspond to the sex chromosomes X and dX. True synapsis is abnormally restricted (43.3%) between these sex chromosomes; on the other hand, self-synapsis of both the X and dX chromosomes is frequent (60%). Single, nonsynapsed axes or axial segments are thickened. Strong chromatin condensation occurs around nonsynapsed axes or axial segments, giving many of these sex pairs an appearance similar to an XY body ("sex vesicle"). The minor gonosome axis differs from that of the Y chromosome of male meiosis, as the former is shorter (relative to the X) and has a different synaptic behavior. In 17 metaphases I from XdX variant females, only heteromorphic, end-to-end joined sex pairs were observed. These variant females differ from the variant females of the wood lemming Myopus schisticolor in several respects, but a similar mechanism seems to be prevalent in other species of the genus Akodon. Self-synapsis of unequal gonosomes in oocytes is assumed as an escape from functional deterioration, following the hypothesis put forward by others.     

The morphological sequence of XY pairing in the Norway rat Rattus norvegicus

The sequence of XY pairing at meiotic prophase in the Norway rat, Rattus norvegicus, has been studied in spread preparations of spermatocytes obtained from pubertal males. As in most mammals, sex chromosome pairing is delayed in relation to that of the autosomes. At one stage in pachytene, the Y is fully paired in synaptonemal complex association with about one-third of the X. Observation in spread preparations at pachytene and diplotene and in air-dried metaphase I preparations indicates that the long arm of the Y pairs with the short arm of the X. Pairing of the Y with both ends of the X is seen in about 4% of pachytene spermatocytes. The possibility that XY pairing in the rat may be nonhomologous (Ashley 1983) is considered, and the view is expressed that the XY synaptonemal complex may be incomplete in fine structural detail, thus not providing for the effective pairing required in true reciprocal recombination. The same mechanism that excludes crossing over from heterochromatic regions of autosomes may also operate to minimize or prevent crossing over in the sex pair of mammals.     

The meiotic structure and behavior of the strongly heteromorphic X/Y sex chromosomes of neotropical plethodontid salamanders of the genus Oedipina

Plethodontid salamanders in the genus Oedipina are characterized by a strongly heteromorphic sex-determining pair of X/Y chromosomes. The telocentric X chromosome and the subtelocentric Y chromosome are clearly distinguished from the autosomes and their behavior during meiosis can be sequentially followed in squash preparations of spermatocytes. In Oedipina the sex chromosomes are not obscured by an opaque sex vesicle during early meiotic stages, making it possible to observe details of sex bivalent structure and behavior not directly visible in other vertebrate groups. The sex chromosomes can first be distinguished from autosomal bivalents at the conclusion of zygotene, with X and Y synapsed only along a short segment at their non-centromeric ends, forming a bivalent that contrasts sharply with the completely synapsed autosomes. During pachytene, the XY bivalent becomes progressively shortened and more compact, disappearing as a visible structure when pachytene progresses into the diffuse stage of male meiosis. Diplotene bivalents gradually emerge from the diffuse nuclei, presumably by the return of the loops of chromatin into their respective chromomeres. During early diplotene, the X/Y bivalent is clearly visible with a single chiasma within the synapsed segment. This chiasma is terminalized by first meiotic metaphase with the X and Y appearing either in end-to-end synaptic contact or as univalents separated at opposite poles relative to the equatorially distributed autosomal bivalents. In C-banded preparations, the Y is entirely heterochromatic while the X contains a large centromeric C-band and another block of heterochromatin located at the telomeric end, in the region of synapsis with the Y. We find no cytological evidence of dosage compensation, such as differential staining of the X chromosomes or Barr bodies, in mitotic or interphase cells from female animals.     

Mitotic and meiotic analysis in Arctocephalus australis (Otariidae)

The karyotype with C-, G- and NOR-banding of Arctocephalus australis is reported for the first time. The chromosomal number is 2n = 36. The X chromosome, identified in G-banded metaphases from males, is metacentric and the Y chromosome is a minute chromosome, also metacentric. Pachytene spermatocytes were used for synaptonemal complexes analysis with a surface spreading technique. A total of 17 autosomal synaptonemal complexes are observed plus the XY pair. During early pachytene, the X and Y axes are thickened and remain unpaired. As pachytene advances, a short SC is formed between the gonosomes, as it is common among eutherian mammals. The particular asymmetrical appearance of the synaptonemal complex in the sex pair is described and compared to other cases among mammals.     

The relationship between chromosomes and axes in the chiasmatic XY pair of the Armenian hamster (Cricetulus migratorius)

The XY pair of the Armenian hamster has been studied in spreads and in three-dimensional reconstructions during the main stages of first meiotic prophase and metaphase I. The general pattern of the axes is similar to that of other mammals. There is a differential and a common region. In the latter a synaptonemal complex (SC) is formed by the pairing of the axes. This SC is longer than in other mammals. Heteropycnosis in the differential region is mirrored by differential chromatin packing at the ultrastructural level. The differential regions of the X and Y chromosomes can be identified both at the light and at the electron microscope level. The location of the axes at the interchromatid space in the differential region has been established. The visualization of the axes with the light microscope is facilitated by their bulgings at the beginning of mid-pachytene. These intermittent deformities change into a coiled and thinner axis during mid-pachytene. A chiasma originates in the common region of the XY body and it is seen near the ends of the sex chromosomes at diakinesis and metaphase I. The ultrastructure of this chiasmatic region is similar to that of autosomal chiasmata in the mouse. The axes separate from each other and leave a remaining piece of SC in which the central space is replaced by dense fibrillar material. During metaphase I the ultrastructure of this chiasmatic region cannot be identified because of the partial loss of the marker axes.     

六种鱼的精母细胞联会复合体的电镜观察

我们以界面铺张——硝酸银染色技术,对鲈形目三种鱼(尼罗罗非鱼、莫桑比克罗非鱼、刺鳅)和鲤形目(鱼句)亚科三种鱼(花(鱼骨)、黑鳍鳈、麦穗鱼)的精母细胞联会复合体进行了电镜观察研究。系统考察了鱼类常染色体SC的亚显微结构、形成过程和配对行为,比较分析了刺鳅的性染色体SC的异配形态和行为,并绘制了鲈形目三种鱼的SC组型模式图。     

Autosomal synaptonemal complexes and sex chromosomes without axes in Triatoma infestans (Reduviidae; Hemiptera)

Meiotic and somatic cells at interphase in Triatoma infestans are characterized by the formation of a large chromocenter, which was assumed to contain the whole of the three large pairs of autosomes and the sex chromosomes. Observations with C-banding techniques show that the chromocenter is formed only by the terminal and subterminal heterochromatic blocks of the three large pairs of autosomes and the sex chromosomes. During pachytene the two largest autosomal pairs loop on themselves and their condensed ends form the chromocenter, together with the single heterochromatic end of the third autosomal pair. The X and Y chromosomes seem to associate with these condensed ends by their affinity for C-heterochromatin. During a very short pachytene stage, bivalents and synaptonemal complexes (SCs) are observed. Pachytene is followed by a very long diffuse stage, during which SCs are disassembled, multiple complexes aggregate on the inner face of the chromocenter and finally all complexes disappear and a dense material is extruded to the cytoplasm through the annuli. The 3-dimensional reconstruction of early pachytene chromocenters show 3 SCs entering and tunnelling the chromocenter, while during mid-pachytene 4 SCs enter this mass and a 5th SC is in a separate small mass. The looping of a whole SC which has both ends in the chromocenter was shown by the reconstructions. These data are interpreted as the progressive looping of the two largest bivalents during pachytene, forming finally the association of 5 bivalent ends corresponding to the 5 C-banding blocks of the large autosomal pairs. No single axis or SC that could be ascribed to the sex chromosomes was found. This agrees with the pachytene microspreads, which show only 10 SCs corresponding to the autosomal bivalents. The X and Y chromosomes are enclosed in the chromocenter, as shown by the unravelling chromocenters at diplotene-diakinesis. Thus the sex chromosomes do not form axial condensations, and this fact may be related to the ability of the X and Y chromosomes to divide equationally at metaphase I. SCsThis paper is dedicated to the memory of the late Professor Francisco A. Saez     

Sex chromosome-autosome translocations in the leaf-nosed bats, family Phyllostomidae. II. Meiotic analyses of the subfamilies Stenodermatinae and Phyllostominae

Analyses of meiotic pairing and synaptonemal complexes of the composite sex chromosomes of male phyllostomid bats with X-autosome or X- and Y-autosome translocations were performed using Giemsa and silver staining procedures. Typical mammalian sex vesicles were absent in all species analyzed. Stenodermatine species with X-autosome translocations possessed an open ring and tail configuration of the XY1Y2 trivalent. Species with both X- and Y-autosome translocations possessed a closed ring and tail configuration of the neo-XY bivalent. In both cases, the tail represented the autosomal short arm of the X paired with its homologue, either the Y2 in XY1Y2 species or the autosomal arm of the composite Y in neo-XY species. Autosomal pairing of the composite sex bivalent in neo-XY species replaced an association between the original X and Y in late prophase I. The absence of a sex vesicle, the unusual pairing configurations of the composite sex chromosomes, and the presumed absence of meiotic nondisjunction in these species is discussed in light of current hypotheses of sex chromosome behavior in male gametogenesis in mammals.     

Morphology and behavior of the sex chromosomes in meiosis in four vole species of the genus Microtus]

Morphology and behaviour of the X and Y chromosomes of four species of genus Microtus were studied at pachytene, metaphase I and meiotic metaphase. The X chromosomes of the species varied with respect to their size and location of heterochromatic blocks. The axes of X and Y chromosomes of these species as well as Microtus agrestis never formed true synaptonemal complexes at any sub-stage of the pachytene. They approached each other at the start of the pachytene throughout to metaphase I, getting situated closely. At the end of the pachytene, they formed sex vesicle. The X and Y chromosomes kept their proximity during metaphase I, but never formed true bivalents. It is suggested that lack of synapsis of the X and Y chromosomes in the genus Microtus is the final step of evolutionary trend to reduction of the size of the pseudo-autosomal region. The abolition of restrictions on homology between the X and Y chromosomes is supposed to be a cause for the fast divergence in morphology of sex chromosomes in the genus.     

The effect of heterochromatin on synapsis of the sex chromosomes of Peromyscus (Rodentia, Cricetidae)

The pairing behavior of the sex chromosomes in male and female individuals representing seven species of Peromyscus was analyzed by electron microscopy of silver-stained zygotene and pachytene configurations. Six species possess submetacentric or metacentric X chromosomes with heterochromatic short arms. Sex-chromosome pairing in these species is initiated during early pachynema at an interstitial position on the X and Y axes. Homologous synapsis then progresses in a unidirectional fashion towards the telomeres of the X short arm and the corresponding arm of the heterochromatic Y chromosome. The distinctive pattern of synaptic initiation allowed a late-synapsing bivalent in fetal oocytes to be tentatively identified as that of the X chromosomes. In contrast to the other species, Peromyscus megalops possesses an acrocentric X chromosome and a very small Y chromosome. Sex-chromosome pairing in this species is initiated at the proximal telomeric region during late zygonema, and then proceeds interstitially towards the distal end of the Y chromosome. These observations suggest that the presence of X short-arm heterochromatin and corresponding Y heterochromatin interferes with late-zygotene alignment of the pairing initiation sites, thereby delaying XY synaptic initiation until early pachynema. The pairing initiation sites are conserved in the vicinity of the X and Y centromeres in Peromyscus, and consequently the addition of heterochromatin during sex-chromosome evolution essentially displaces these sites to an interstitial position.     

Immunocytochemical labelling of the kinetochore of human synaptonemal complexes,and the extent of pairing of the X and Y chromosomes

An immunocytochemical method was used to label the kinetochores on human synaptonemal complexes. Synaptonemal complex spreads were labelled with autoimmune CREST serum, followed by a second antibody labelled with colloidal gold, and examined by electron microscopy. Clusters of gold particles were found at discrete sites which were identified as kinetochores on the autosomal synaptonemal complexes, as well as on the XY pair. This method was used to investigate the extent of pairing of the human X and Y chromosomes at pachytene. Our observations confirm earlier work, based purely on measurements, that the pairing of the sex chromosomes sometimes extends beyond the centromere of the Y chromosome into the long arm. At the same time we showed that the centromeric indices of the X and Y at pachytene are highly variable, so that measurements alone are not sufficient to estimate the degree of pairing of the sex chromosomes.     

Deficiency of X and Y chromosomal pairing at meiotic prophase in spermatocytes of sterile interspecific hybrids between laboratory mice (Mus domesticus) and Mus spretus

The normal association between the X and Y chromosomes at metaphase I of meiosis, as seen in air-dried light microscope preparations of mouse spermatocytes, is frequently lacking in the spermatocytes of the sterile interspecific hybrid between the laboratory mouse strains C57BL/6 and Mus spretus. The purpose of this work is to determine whether the separate X and Y chromosomes in the hybrid are asynaptic, caused by failure to pair, or desynaptic, caused by precocious dissociation. Unpaired X-Y chromosomes were observed in air-dried preparations at diakinesis, just prior to metaphase I. Furthermore, immunocytology and electron microscopy studies of surface-spread pachytene spermatocytes indicate that the X and Y chromosomes frequently fail to initiate synapsis as judged by the failure to form a synaptonemal complex between the pairing regions of the X and Y Chromosomes. Several additional chromosomal abnormalities were observed in the hybrid. These include fold-backs of the unpaired X or Y cores, associations between the autosome and sex chromosome cores, and autosomal univalents. The occurrence of abnormal autosomal and XY-autosomal associations was also correlated with cell degeneration during meiotic prophase. The primary breakdown in hybrid spermatogenesis occurs at metaphase I (MI), with the appearance of degenerated cells at late MI. In those cells, the X and Y are decondensed rather than condensed as they are in normal mouse MI spermatocytes. These results, in combination with the previous genetic analysis of spermatogenesis in hybrids and backcrosses with fertile female hybrids, suggest that the spermatogenic breakdown in the interspecific hybrid is primarily correlated with the failure of XY pairing at meiotic prophase, asynapsis, followed by the degeneration of spermatocytes at metaphase I. Secondarily, the failure of XY pairing can be accompanied by failure of autosomal pairing, which appears to involve an abnormal sex vesicle and degeneration at pachytene or diplotene.by C. Heyting     

On the nature and extent of XY pairing at meiotic prophase in man

Evidence is presented that pairing between the human X and Y chromosomes could be more extensive at early pachytene than has previously been supposed and could involve even the entire euchromatic portion of the Y chromosome. Following desynapsis over the major part of the X and Y axes, a small paired segment of Xp and Yp remains into late pachytene. Association between the distal tips of Xq and Yq can also be observed in about one half of the spermatocytes examined. A hypothesis linking meiotic pairing to early replicating sites along the chromosomes is proposed.     

Electron microscopic analysis of synaptonemal complexes of male laboratory mice exposed during the period of embryogenesis in the vicinity of the Chernobyl Nuclear Power Station]

Four male mice were shown to have decreased fertility among 74 laboratory male mice exposed in the Chernobyl APP area during embryogenesis. Electron microscopic analysis of synaptonemal complexes (SC) of at zygotene-diplotene and light-optic analysis of chromosome aberrations at diakinesis-metaphase 1 demonstrated the presence of interchromosome translocations in autosomal chromosomes, and also intrachromosome translocations in two of four animals. The frequency of chromosome translocations in SC preparations was on the average 1.8 times higher than their frequency at the diakinesis-metaphase 1 stage. High percentage of cells in which associations of sex bivalent axes (XY) with the axes of autosomal bivalents were observed in the SC preparations and reduced fertility in the animals studied confirmed the Forejt's hypothesis that such associations caused the arrest of cells at pachytene and, as a consequence, disturbed the process of embryogenesis.