Sex chromosome pairing during male meiosis in marsupials

The pairing of the sex chromosomes at pachytene has been examined in twenty-two species of Australian marsupials, including four with complex sex chromosome systems. The axial elements of the sex chromosomes associate in all but one species. However, no synaptonemal complex has been observed between the axes of the X and Y chromosome in any of the examined species. Both the type of association between the sex chromosome axes, and the structural modifications of these axes are conserved within taxonomic groupings. In three species with complex sex chromosome systems, the t(XA), Y, A trivalents do not have a favoured relative orientation of the axes of the Y and A chromosomes, whereas in a fourth species with a t(XA₁), t(A₂YA₂), A₂ system the t(XA₁) and A₂ axes are in a cis arrangement with each other.     

DNA methylation analysis of ade novo balanced X;13 translocation in a girl with abnormal phenotype: evidence for functional duplication of the whole short arm of the X chromosome

We report on a 13-month-old girl showing dysmorphic features and a delay in psychomotor development. She was diagnosed with a balancedde novo translocation 46, X, t(X;13)(p11. 2;p13) and non-random inactivation of the X chromosome. FISH analysis, employing the X chromosome centromere andXIST-region-specific probes, showed that theXIST locus was not involved in the translocation. Selective inactivation of paternal X, which was involved in translocation, was revealed by the HUMARA assay. The pattern of methylation of 5 genes located within Xp, which are normally silenced on an inactive X chromosome, corresponded to an active (unmethylated) X chromosome. These results revealed that in our proband the X chromosome involved in translocation (Xt) was preferentially inactivated. However, genes located on the translocated Xp did not includeXIST. This resulted in functional Xp disomy, which most probably accounts for the abnormal phenotype in our patient.     

Three dimensional reconstruction of human pachytene spermatocyte nuclei of a 17;21 reciprocal translocation carrier: study of XY-autosome relationships

Summary A study of XY-autosome relationships at the pachytene stage in an infertile 17–21 reciprocal translocation carrier was undertaken by means of three dimensional reconstruction. Synaptonemal complexes and the sex vesicle were analysed on electron microscopic serial sections and the reconstruction was performed on transparent sheets and on a Samba 2000 (Alcatel TITN) image analysis system. All asynapsed segments were entirely included in the sex vesicle, the chromatin fibre of the autosomes and sex chromosomes being tightly intermingled. In one nucleus, the four arms of the quadrivalent were paired, except around the breakpoints where an interstitial asynapsis was observed. In the other nuclei, a terminal asynapsis involving one or two arms of the quadrivalent was found. In the sex vesicle, autosomal asynapsed segments showed the same morphological characteristics as those of X and Y chromosomes. This observation agrees with the hypothesis of the extension of gene inactivation from sex chromosomes to autosomes.     

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.     

Analysis of male meiotic "sex body" proteins during XY female meiosis provides new insights into their functions

During male meiosis in mammals the X and Y chromosomes become condensed to form the sex body (XY body), which is the morphological manifestation of the process of meiotic sex chromosome inactivation (MSCI). An increasing number of sex body located proteins are being identified, but their functions in relation to MSCI are unclear. Here we demonstrate that assaying male sex body located proteins during XY female mouse meiosis, where MSCI does not take place, is one way in which to begin to discriminate between potential functions. We show that a newly identified protein, "Asynaptin" (ASY), detected in male meiosis exclusively in association with the X and Y chromatin of the sex body, is also expressed in pachytene oocytes of XY females where it coats the chromatin of the asynapsed X in the absence of MSCI. Furthermore, in pachytene oocytes of females carrying a reciprocal autosomal translocation, ASY associates with asynapsed autosomal chromatin. Thus the location of ASY to the sex body during male meiosis is likely to be a response to the asynapsis of the non-homologous regions [outside the pseudoautosomal region (PAR)] of the heteromorphic X-Y bivalent, rather than being related to MSCI. In contrast to ASY, the previously described sex body protein XY77 proved to be male sex body specific. Potential functions for MSCI and the sex body are discussed together with the possible roles of these two proteins.     

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.     

Ultrastructure and behavior of the achiasmatic,telosynaptic XY pair of the sand rat (Psammomys obesus)

The behavior of the X and Y chromosomes in somatic and testicular cells of the sand rat (P. obesus) has been investigated with light and electron-microscope procedures. The Y chromosome has been identified as the fourth longest of the complement, both by C-banding and by its meiotic behavior. The X chromosome is the longest of the complement and carries two major C-heterochromatic blocks, one in the distal part of the long arm and the other forming most of the short arm. During presynaptic stages in spermatocytes, separate C-heterochromatic blocks, representing the sex chromosomes, are observed in the nuclei. An XY body is regularly formed at pachytene. During first meiotic metaphase the X and Y chromosomes show variable associations, none of them chiasmatic. Second meiotic metaphases contain, as in other mammals, a single sex chromosome, suggesting normal segregation between the X and the Y. — Electron microscopic observations of the autosomal synaptonemal complexes (SCs) and the single axes of the X and Y chromosomes during pachytene permit accurate, statistically significant identification of each of the largest chromosomes of the complement and determination of the mean arm ratios of the X and Y axes. The X and Y axes always lie close to each other but do not form a SC. The ends of the X and Y axes are attached to the nuclear envelope and associate with each other in variable ways, both autologously (X with X or Y with Y) and heterologously (X with Y), with a tendency to form a maximum number (four) of associated ends. Analysis of 36 XY pairs showed no significant preference for any single specific attachment between arm ends. The eighth longest autosomal bivalent is frequently partially asynaptic during early pachytene, and only at that time is often near or touching one end of the X axis. — It is concluded that while axis formation and migration of the axes along the plane of the nuclear envelope proceed normally in the X and Y chromosomes, true synapsis (with SC formation) does not occur because the pairing region of the X chromosome has probably been relocated far from the chromosome termini by the insertion of distal C-heterochromatic blocks.     

Deletion of specific sequences or modification of centromeric chromatin are responsible for Y chromosome centromere inactivation

Summary Stable dicentric chromosomes behave as monocentrics because one of the centromeres is inactive. The cause of centromere inactivation is unknown; changes in centromere chromatin conformation and loss of centromeric DNA elements have been proposed as possible mechanisms. We studied the phenomenon of inactivation in two Y centromeres, having as a control genetically identical active Y centromeres. The two cases have the following karyotypes: 45,X/46,X,i(Y)(q12) and 46,XY/ 47,XY,+t(X;Y)(p22.3;p11.3). The analysis of the behaviour of the active and inactive Y chromosome centromeres after Da-Dapi staining, CREST immunofluorescence, and in situ hybridization with centromeric probes leads us to conclude that, in the case of the isochromosome, a true deletion of centromeric chromatin is responsible for its stability, whereas in the second case, stability of the dicentric (X;Y) is the result of centromere chromatin modification.     

Meiotic studies in mice carrying the sex reversal (Sxr) factor

A sex reversal factor (Sxr) that causes mice having apparently normal X chromosomes to become phenotypically male is transmitted in an autosomal pattern. The origin of the Sxr factor is still unknown. It seems most likely that it has originated from an autosomal gene mutation or is the result of a translocation of part of the Y chromosome to one of the autosomes. Chromosomes from four XY and six XO mice carrying this sex reversal factor were examined in the diakinesis stage of meiosis. The following unusual observations were noted: (1) in XY males carrying the Sxr factor, the X and Y chromosomes were separated more often than in controls. (2) The Y chromosome tends to be closer to an autosome when the X and Y are separate than when the X and Y are attached. (3) A chromosome fragment was present in 4/226 cells from two XO males and a single cell from an XY, Sxr carrier. Although there is no direct evidence, these observations seem to favor the possibility that the Sxr factor involves a chromosomal rearrangement rather than a single gene mutation.     

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

Mitotic analyses using RBA- and C-banding were performed on Stenodermatine bats with X-autosome (XY1Y2) and X- and Y- autosome (neo-XY) translocations. RBA-banded metaphases of females revealed differential replication of the inactive X chromosome. An early replicating band comprises the short arm of the X, and an intermediate replicating band is located interstitially on the long arm. The early replicating short arm has a homologous counterpart either in the form of a free autosome (the Y2) or as part of the Y. Both the "autosomal" short arm of the X and its homologue fused to the Y are C-band negative and behave autonomously from the remainder of the sex chromosomes. They are separated from X and Y chromatin by centromeric heterochromatin which presumably acts as a barrier. The intermediate replicating region of the long arm of the X is also present in the subfamily Phyllostominae. In both subfamilies this region lacks a homologous counterpart. However, it may also represent a translocated autosome which, unlike the short arm of the X, is not separated from the inactive X by centromeric heterochromatin. Its intermediate replication time may represent a retarded replication due to its juxtaposition to late replicating X chromatin. These data are discussed in light of the theory of the evolution of sex chromosome heteromorphism, specifically as it applies to mammals.     

A complex sex-chromosome system in the hare-wallaby Lagorchestes conspicillatus Gould

Among specimens of the spectacled hare-wallaby Lagorchestes conspicillatus Gould (Marsupialia, family Macropodidae) 4 males had 15 chromosomes and 2 females 16 chromosomes. The sex chromosomes are X₁X₁X₂X₂ in the female and X₁X₂Y in the male, the Y being metacentric and both X chromosomes are acrocentric. In about 96% of sperm mother cells at meiosis the sex chromosomes form a chain trivalent and in more than 99% of these this orients convergently so that the X₁ and X₂ move to the same pole. Evidence is presented that L. conspicillatus has evolved from a form with 22 chromosomes including a small X and a minute Y. Autoradiographic studies show that the proximal fifth of the X₁ chromosome replicates late. This is probably the ancestral X chromosome which has been translocated to an autosome. The fate of the original Y is obscure but an hypothesis is proposed that it forms the centromeric region of the Y. A single male had 14 chromosomes and was heterozygous for a translocation involving the centric fusion of two acrocentric autosomes. In about 30% of sperm mother cells the autosomal trivalent did not disjoin regularly but, despite this, all secondary spermatocytes observed at metaphase 2 had balanced complements of chromosomes. It is assumed that unbalanced secondary spermatocytes died before reaching metaphase.     

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.     

Y-autosome translocation and infertility: usefulness of molecular, cytogenetic and meiotic studies

An apparently balanced reciprocal translocation 46,X,t(Y;6) (q11.23 ∼ q12;p11.1) was observed in an infertile man with severe oligozooteratozoospermia. Different mitotic chromosome banding patterns were performed and fluorescence in situ hybridization indicated a breakpoint in the fluorescent Yq heterochromatin. Molecular genetic deletion experiments for the azoospermia factor region in distal Yq11 showed the retention of the DAZ gene and meiotic pairing configurations suggested that the man’s infertility could be due to the pairing behaviour of the Y;6 translocation chromosome with the X chromosome visualised by synaptonemal complex analysis at the electron microscopy level. The morphological appearance of the normal chromosome 6 and the Y;6 translocated chromosome included in the compartment of the sex vesicle may allow an explanation of the degeneration of most spermatocytes after the pachytene stage. Received: 1 August 1997 / Accepted: 25 September 1997     

Identification and patterns of synapsis of the autosomally translocated Y-chromosome of the Indian mongoose,Herpestes auropunctatus (Hodgson)

The multiple sex chromosome system, X₁X₂Y /X₁X₁X₂X₂, in the small Indian mongoose, Herpestes auropunctatus, results from a translocation of a part of Y chromosome to an autosome. It is not possible to distinguish the autosome which harbours the Y chromosome element in the somatic complement. By employing the surface-spreading technique to prophase I meiocytes we have identified the region to which the Y chromosome has been translocated as the short arm of chromosome 9 which is a subtelocentric chromosome. This Y chromosome component lacks heterochromatin and no sex vesicle is organised during meiotic prophase. This suggests to us that Y heterochromatin in mammals may be required for the production of a sex vesicle.We take great pleasure in dedicating this paper to our revered teacher Prof. S.P. Ray-Chaudhuri, who initiated us to the field of Cytogenetics, on the occasion of his 75th birth day     

A unique dicentric X;Y translocation with Xq and Yp breakpoints: cytogenetic and molecular studies.

A 32-year-old woman presented with secondary amenorrhea and infertility. She was of normal height and her breasts were well developed, but she had streak gonads; there were no signs of virilization, and she showed no somatic stigmata of Turner syndrome. Chromosome analysis revealed a dicentric X;Y translocation with Xq and Yp breakpoints. Centromeric banding demonstrated a Y centromere and a "suppressed" X centromere. The karyotype of the patient was interpreted as 46,X,t(X;Y)(q22;p11). The Yp breakpoint was confirmed by DNA-hybridization studies with six probes detecting Y-specific sequences. These DNA-hybridization studies were consistent with the presence of the long arm, centromere, and much of the proximal short arm of the Y. The Y-DNA studies of this female also revealed the absence of the distal short arm of the Y chromosome, to which the testis-determining factor has previously been localized.     

Synaptonemal complex analysis of X-7 translocations in male mice: R2 and R6 quadrivalents

Synaptonemal complexes of surface-spread spermatocytes of mice heterozygous for reciprocal translocations R2 or R6 between the X-chromosome and chromosome 7 were examined by light and electron microscopy (EM). Measurements of the lengths of all chromosome axes involved in the translocation configurations and of the extent of synapsis were used to calculate the position of the break points of the two translocations. The breaks for R2 were determined to be at 62% of the 7 as measured from the centromere, and at 27% of the X. Quadrivalents were formed almost exclusively. The break points for R6 were calculated to be at 30% of the 7 as measured from the centromere, and at 75% of the X. Although in R6 the break in the X lies within the potential pairing region of the sex chromosomes, univalent Ys were rarely observed (6%). The EM sample of 76 nuclei contained: 42% quadrivalents, 52% heteromorphic bivalents, 4% trivalent plus Y univalent, and 2% X7-7 bivalent plus two univalents (7X and Y). Nonhomologous synapsis occurred in the quadrivalents of both R2 and R6. In R6 nonhomologous synapsis of the X portion of the 7X with the 7 involved up to 14% of the length of the 7. Methods are discussed for determining the position of the break points in the presence of nonhomologous synapsis. It is proposed that the high percentage of bivalents is due to premature desynapsis of the 7X from the 7 and that the X portion of the 7X axis confers its property of premature desynapsis on that portion of the 7 to which it is attached.     

Karyotype,heterochromatin content and meiotic features ofPoecilocera picta (Orthoptera,Acrididae)

Cytological study of three distinctly separated populations ofPoecilocera picta revealed a chromosome number of 2N = 18 + XO/ XX. Except for the hemizygosity of a procentric heterochromatic block in the M₆ pair of the Bangalore population, the basic karyotype of the three populations is markedly similar. The autosomal karyotype formula is 2Lt + 4Mt + 1 Mst + 2S st and the telocentric X chromosome is the longest of the complement. All bivalents at pachytene carried procentric heterochromatic blocks. The M₄ is the nucleolus organiser with the NOR region situated interstitially but proximal to the centromere. About 11 μm (4%) of the total (290 μm) autosomal pachytene complement is heterochromatic; a major portion of it is contributed by the S₉ pair which is mostly heterochromatic. Chiasmata are localized proximally and distally and in the S₉ pair their formation is confined to the short procentric euchromatic segment of the long arm. Female meiosis did not reveal any chromomere pattern at pachytene and, unlike in the male, the sex bivalent in the female is indistinguishable from the autosomal bivalents. G- and C-banding patterns in males showed procentric bands in all the chromosomes. In addition there are eight telomeric and two interstitial bands which are C negative. The S₉ pair showed only two bands. The G-banding pattern of the sex chromosome in meiosis showed only a centric band while the heterochromatic body of the facultatively heterochromatic X remained G negative.     

Cytogenetics of ticks (Acari: Ixodoidea)

Summary Prior to this paper there have been no reports of a multiple sex chromosome mechanism operative in any tick. The present paper deals with two species of Ixodidae, Amblyomma moreliae and Amblyomma limbatum that exhibit an X₁X₁X₂X₂:X₁X₂Y type of sex chromosome mechanism. Cells from males of both species show nine bivalents plus one sex trivalent. Eleven bivalents were observed in one female A. moreliae. The sex trivalent probably evolved through reciprocal translocation from a system that included ten autosomal bivalents and one sex univalent (the system found in most ixodid species). As a result of the translocation, there are now two X chromosomes (X₁ and X₂) segregating from an unaltered autosome, the neo-Y. A large X chromosome is characteristic of many ticks; in this instance the reciprocal translocation did not change appreciably its relative size.The opinions and assertions contained herein are the private ones of the author and are not to be construed as official or reflecting the views of the Navy Department or the Naval service at large.This study was begun during the tenure of a North Alantic Treaty Organization (National Science Foundation) Postdoctoral Fellowship.