XY pair associates with the synaptonemal complex of autosomal male-sterile translocations in pachytene spermatocytes of the mouse (Mus musculus)

Analysis of the chromosome behaviour at pachytene has been performed by means of the silver staining technique visualizing the synaptonemal complexes (SCs) in male mice heterozygous for the male-sterile translocations T(5;12)31H, T(16;17)43H and T(7;19)145H, respectively. The T(9;17)138Ca male heterozygotes and T43H/T43H homozygous males were used as fertile controls. The sterile mice displayed a high frequency (about 60%) of pachytene spermatocytes with autosomal translocation configuration located in close vicinity of the XY pair. The dense round body (XAB), normally located near the X-chromosome axis in fertile males, exhibited abnormal affinity to translocation configuration in the sterile translocation heterozygotes. The incomplete synapsis of autosomes involved in translocation configuration was observed in more than 70% of the pachytene spermatocytes with the male-sterile translocations but in less than 20% of the cells from T138Ca fertile male.s. A hypothesis relating the spermatogenic arrest of carriers of male-sterile rearrangements to the presumed interference with X chromosome inactivation in male meiosis is discussed.     

Genetic analysis of fertility of male mice with various t-haplotypes

Fertility of 47 mouse males carrying various combinations of lethal, t-haplotypes (t6/tw18, t12/tw18, Tw73/tw12, tw5/tw18, t6/dt5, t12/tw12, tw5/twPa-1, tw18/twPa-1, tw5/tw12) was studied in crosses with females of different genotypes. The t-haplotypes studied belong to 7 main groups of complementation. The presence of at least two factors of fertility in the t-complex was revealed. The influence of female genotype on the degree of male fertility was also demonstrated. The data presented confirm that different combinations of lethal complete t-haplotypes exhibit sterility, with the exception of t8/tw18 compound.     

Genetic analysis of complementation of t-haplotypes in mice

Complementation for viability of the compounds tx/ty in complete lethal haplotypes t6, t12, tw5, tw12, tw18, tw73, twPa-1 belonging to the seven basic complementation groups was studied. In the overwhelming majority of crosses a lack of tx/ty offspring was revealed but there is also evidence of non-complete complementation of the tx/ty haplotypes, except for T/tw73XT/tw12, T/tw18XT/t12, T/tw73XT/tw18 crosses in which the level of complementation for viability exceeds 100%. The maternal effect for compound variability was significantly revealed in 9 out of the 19 matings. In addition, reciprocal matings of heterozygotes T/txXT/ty were tested for fertility. In two cases, the maternal effect was also revealed for this character. Comparison of data on viability of the compounds and the fertility of the original strains gives grounds to believe that the tx/ty haplotypes are not completely recessive and partially manifest themselves in the heterozygotes, decreasing fertility in a number of matings. In some cases, these t-haplotypes yield an effect, close to negative complementation, which may be associated with the interaction of the proteins of t-haplotypes.     

A pachytene analysis of two male-fertile paracentric inversions in chromosome 1 of the mouse and in the male-sterile double heterozygote

Meiotic studies have been made at pachytene on two paracentric inversions in chromosome 1 of the mouse. Surface-spread preparations of primary spermatocytes have been analysed at the light microscope level in males heterozygous for the inversions In(1)1Rk and In(1)12Rk and in the double heterozygote In(1)1RK/In(1)12Rk. In singly heterozygous form, neither inversion produces any serious effect on male fertility. In the double heterozygote, spermatogenesis is arrested in the majority of cells at the spermatocyte stage and males are rendered totally sterile by azoospermia. In the double heterozygote, a complex loop, indicating the inversion bivalent, is found in 90% of pachytene cells analysed. In the In(1)1Rk/+ heterozygote, a looped bivalent was seen in 47 per cent of pachytene cells but in In(1)12Rk/+ no cells containing loops could be found. -80% of pachytene spermatocytes from the In(1)1Rk/In (1)12Rk double heterozygote showed apposition of the inversion bivalent to the sex bivalent. Such an association was rarely seen in pachytene cells of either of the fertile single heterozygotes. Spermatogenic failure in the double heterozygote may be related to interference, by the inversion bivalent, with X chromosome inactivation at meiotic prophase.     

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.     

Effect of heterozygosity for insertions of homogeneously stained regions in chromosome 1 of the house mouse on synapsis in meiotic prophase

Electron microscope analysis of surface-spread synaptonemal complexes (SC) in oocytes and spermatocytes from double cis heterozygotes for Is(HSR; 1C5)1Icg and Is(HSR; 1E3)2Icg was carried out. Aberrant chromosomes were isolated from the feral population of Mus musculus musculus of Novosibirsk. They contain homogeneously stained regions of total length of about 30% of Chr 1 mitotic metaphase. Heteromorphic bivalents of Chr1 with different lengths of the lateral elements of SC and the loop in the intermedial position were revealed in 4.4% spermatocytes and 20% oocytes of heterozygous animals. The loop size depends on the stage of meiosis: it is maximal at late zygotene and decreases up to disappearance during pachytene.     

Synaptonemal complex analysis in spermatocytes and oocytes of turbot, Scophthalmus maximus (Pisces, Scophthalmidae).

A surface-spreading synaptonemal complex (SC) technique was used to analyze spermatocytes and oocytes of turbot (Scophthalmus maximus) to visualize the process of chromosome synapsis. The total SC length was 205 +/- 12 microm in males and 172 +/- 29 microm in the only female analyzed. A representative SC karyotype of turbot was obtained. Each SC showed lateral elements of equal length. No bivalent exhibiting atypical synaptic behaviour that could be associated with heteromorphic sex chromosomes was observed, either in males or in the female. The DNA content of turbot was evaluated in eight individuals of both sexes by flow cytometry analysis. The 2C mean DNA content of turbot (1.308 +/- 0.009 pg/cell) was among the lowest observed within fishes. No statistical differences in DNA content were revealed between the sexes [Wilcoxon/Mann-Whitney test; P(W(x) = 0.243)]. The SC/DNA content ratio observed in turbot was the highest reported to date in bony fishes (Osteichthyes).     

Synaptonemal Complex und Chromosomenstruktur in der achiasmatischen Spermatogenese vonPanorpa communis (Mecoptera)

Meiotic prophase in the spermatocytes ofPanorpa communis was studied. There is a proper sequence of meiotic stages in the testes. Therefore the temporal development of chromosome structure and the synaptonemal complex (SC) could be studied exactly. The structure and function of the SC are interpreted in a new model.—The chromosomes have a lambrush form from leptotene to diakinesis. At leptotene each chromatid produces an additional axis of basic protein and RNA. The axis becomes one of the lateral elements of the SC. At pachytene the DNA of the bivalents is separated into three regions: 1. Most of the DNA forms long loops outside the SC. 2. Smaller portions of the DNA filaments are twisted around the lateral elements of the SC. 3. Short DNA loops (called pairing loops) extend into the pairing space. InPanorpa the SC is composed of two lateral elements (chromosome axes), which are connected by equally spaced transverse filaments, a ladder-like central element in the middle of the pairing space and, on each side of the pairing space parallel to the lateral elements, two RNA containing strands. These are regarded as connected RNA copies of the pairing loops and are responsible for the exact pairing of homologous chromosome segments. At diplotene the axes of the sister chromatids separate to form “double complexes” with four lateral elements. The double complexes of the oocytes contain only transverse filaments between the axes of the homologous chromatids. After a short time they disappear again and the homologues separate to form the chiasmatic bivalents. In the spermatocytes all four chromatid axes are connected by transverse filaments. The pairing complex persists until diakinesis, thereby causing the suppression of the diplotene stage in the light microscope. This may be the only reason for the achiasmatic meiosis in the spermatocytes ofPanorpa.     

Synaptonemal complex analysis of mouse chromosomal rearrangements

Electron microscopy of surface-spread spermatocytes from mice heterozygous for a tandem duplication shows the heteromorphic synaptonemal complex (SC) to comprise two lateral elements of unequal length, the longer of which is buckled out in a characteristic loop, representing the unsynapsed portion of the duplication. The loop is a regular feature of late zygotene-early pachytene nuclei; it is longest at these early stages, but, through equalization of the two axes as a consequence of synaptic adjustment, it is replaced by a normal appearing SC at late pachytene. Because equalization, as indicated by a decrease in the percent difference between axes, may begin shortly after completion of synapsis, estimates of duplication segment length are restricted to a sample selected for least adjustment. — Although the mean position of the loop is constant at various pachytene substages, individual positions vary widely from cell to cell, consistent with the behavior expected of a duplication, but not of a deletion or an inversion. The length of the segment that is duplicated is estimated to be 22% of the normal chromosome, the midpoint of the segment is mapped at 0.61 of the chromosome distal to the kinetochore, and the ends of the segment are mapped at 0.50 to 0.72. Measurements of G-banded mitotic chromosomes give comparable values: duplication length, 24%; midpoint, 0.60, and segment ends, 0.48 and 0.71. This agreement constitutes further validation of the SC/spreading method for detecting and analyzing chromosomal rearrangements at pachytene and substantiates the fidelity with which the axes and SCs represent the behavior of chromosomes in synapsis.     

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.     

Analysis of chromosome aberrations on the basis of synaptonemal complexes in the offspring of mice irradiated with gamma-rays

Electron-microscopic analysis of synaptonemal complexes (SC) spread on the surface of hypophase was carried out to study chromosome rearrangements in sterile and semisterile F1 offsprings of mice exposed to gamma-radiation at a dose of 5 Gy. Chromosome rearrangements were microscopically scored at diakinesis - metaphase I in the same animals. SC analysis at pachytene revealed chromosome rearrangements in 63% spermatocytes. Analysis of chromosomes at diakinesis - metaphase I in the same animals only revealed chromosome rearrangements in 32% cells. SC analysis permits detecting chromosome rearrangements undetectable at diakinesis - metaphase I.     

Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries

During mammalian meiotic prophase, homologous chromosomes connect through the formation of the synaptonemal complex (SC). SYCP3 is a component of the lateral elements of the SC. We have generated transgenic mice expressing N- or C-terminal fluorescent-tagged SYCP3 (mCherry-SYCP3 (CSYCP) and SYCP3-mCherry (SYCPC)) to study SC dynamics and chromosome movements in vivo. Neither transgene rescued meiotic aberrations in Sycp3 knockouts, but CSYCP could form short axial element-like structures in the absence of endogenous SYCP3. On the wild-type background, both fusion proteins localized to the axes of the SC together with endogenous SYCP3, albeit with delayed initiation (from pachytene) in spermatocytes. Around 40% of CSYCP and SYCPC that accumulated on the SC was rapidly exchanging with other tagged proteins, as analyzed by fluorescent recovery after photobleaching (FRAP) assay. We used the CSYCP transgenic mice for further live cell analyses and observed synchronized bouquet configurations in living cysts of two or three zygotene oocyte nuclei expressing CSYCP, which presented cycles of telomere clustering and dissolution. Rapid chromosome movements were observed in both zygotene oocytes and pachytene spermatocytes, but rotational movements of the nucleus were more clear in oocytes. In diplotene spermatocytes, desynapsis was found to proceed in a discontinuous manner, whereby even brief chromosome re-association events were observed. Thus, this live imaging approach can be used to follow changes in the dynamic behavior of the nucleus and chromatin, in normal mice and different infertile mouse models.     

Immunocytogenetics. III. Analysis of trivalent and multivalent configurations in mouse pachytene spermatocytes by human autoantibodies to synaptonemal complexes and kinetochores

Mice heterozygous for one or more Robertsonian (Rb) translocation chromosomes have been used to analyze synaptonemal complex (SC) configurations and kinetochore arrangements in trivalents and multivalents. Rb heterozygosity without arm homologies leads to the formation of heteromorphic trivalents in meiosis I; alternating homology of the chromosome arms produces ringlike or chainlike multivalents. Immunofluorescence double-labeling with human antibodies to SCs and kinetochores was performed on surface-spread pachytene spermatocytes. Both Rb bivalents and Rb trivalents clearly showed that metacentrics possess only one centromere. In heteromorphic trivalent SCs, the nonhomologous kinetochores of the two acrocentrics were closely paired in a cis-configuration and juxtaposed opposite the kinetochore of the metacentric; the latter appeared to be an integral part of the longitudinal SC axis. Meiotic multivalents of interpopulation hybrids included up to 36 chromosome arms. In multivalent SCs, the kinetochores always lay together, with the SC arms arranged away from the central centromere cluster. The paracentromeric regions of the Rb chromosomes appeared to remain unsynapsed on both sides of the centromeres. The SC arms were often linked by end-to-end associations. Following desynapsis of the multivalent SC, the kinetochores of the Rb metacentrics showed a highly nonrandom topologic distribution within the nucleus, reminiscent of their arrangement during synapsis.     

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.     

Robertsonian chromosomes and the nuclear architecture of mouse meiotic prophase spermatocytes

Background

The nuclear architecture of meiotic prophase spermatocytes is based on higher-order patterns of spatial associations among chromosomal domains from different bivalents. The meiotic nuclear architecture depends on the chromosome characteristics and consequently is prone to modification by chromosomal rearrangements. In this work, we consider Mus domesticus spermatocytes with diploid chromosome number 2n = 40, all telocentric, and investigate a possible modification of the ancestral nuclear architecture due to the emergence of derived Rb chromosomes, which may be present in the homozygous or heterozygous condition.

Results

In the 2n = 40 spermatocyte nuclei random associations mediated by pericentromeric heterochromatin among the 19 telocentric bivalents ocurr at the nuclear periphery. The observed frequency of associations among them, made distinguishable by specific probes and FISH, seems to be the same for pairs that may or may not form Rb chromosomes. In the homozygote Rb 2n = 24 spermatocytes, associations also mediated by pericentromeric heterochromatin occur mainly between the three telocentric or the eight metacentric bivalents themselves. In heterozygote Rb 2n = 32 spermatocytes all heterochromatin is localized at the nuclear periphery, yet associations are mainly observed among the three telocentric bivalents and between the asynaptic axes of the trivalents.

Conclusions

The Rb chromosomes pose sharp restrictions for interactions in the 2n = 24 and 2n = 32 spermatocytes, as compared to the ample possibilities for interactions between bivalents in the 2n = 40 spermatocytes. Undoubtedly the emergence of Rb chromosomes changes the ancestral nuclear architecture of 2n = 40 spermatocytes since they establish new types of interactions among chromosomal domains, particularly through centromeric and heterochromatic regions at the nuclear periphery among telocentric and at the nuclear center among Rb metacentric ones.     

Chromosome banding and genome compartmentalization in fishes

The diploid chromosome number of the Chinese raccoon dog varies from 54 (no B chromosomes) to 58 (4 B chromosomes). The B chromosomes are totally heterochromatic. An electron microscopic study was made of the synaptonemal complexes (SC) in spermatocytes of these animals. The SC karyotype consists of 27 regular chromosome pairs (autosomes and the sex chromosomes) plus the B chromosomes. The Bs pair effectively with one another at pachytene, but the SC axes of the B chromosomes are much denser than those of the A chromosomes. Depending on the number of Bs, both bivalents and multivalents have been observed. When three B chromosomes are present in a cell, parallel alignment of all three SCs can be seen. Formation of multivalents indicates high homology among these supernumerary heterochromatic chromosomes. Fusiform bulges are found along unpaired regions of all chromosomes which are particularly pronounced in diplotene.     

Prophase of meiosis in human spermatocytes analysed by EM microspreading in infertile men and their controls and comparisons with human oocytes

Summary Observations at the electron microscope (EM) level have been made on 1883 primary spermatocytes from 40 chromosomally normal subfertile men and 566 spermatocytes from 10 fertile controls, using the technique of microspreading. Spermatocytes of infertile men in general showed greater indications of degeneration including higher levels of background silver deposition, nucleolar organising region — XY associations, fragmentation of synaptonemal complexes and overproduction of XY excrescences. A few oligospermic men also showed an immature morphology of the XY pair and/or a reduced extent of XY synapsis. Dissociation of the sex chromosome axes at prophase was found to occur with a much lower frequency than that recorded for separated X and Y chromosomes at metaphase I. In a single spermatocyte, synaptonemal complex formation was observed between Xqter and Yqter, a situation that could enable rare XqYq interchange. A proteinaceous stalked body exists on the Y axis towards its non-pairing end; this structure might have a functional relationship with the gene for spermatogenesis, (AZF), located at the euchromatin/heterochromatin interface. Compared with human oocytes, spermatocytes show fewer anomalies of synapsis, i.e. asynapsed segments or whole axes, non-homologous associations, interchanges, interlocks. These latter data agree well with findings from the mouse.     

家鸡联会复合体的亚显微结构分析

本文以表面铺展——硝酸银染色技术,对家鸡的联会复合体(Syneptonemal Complex,SC)作亚显微结构分析。根据对10个精母细胞和10个卵母细胞SC的测量结果,绘制组型图。发现雌雄家鸡的常染色体的SC组型相同。在精母细胞中,性染色体(ZZ)的行为与常染色体相似。在卵母细胞中,性染色体ZW的长度不同,长轴为Z,短轴为W,两者之间只有部分配对,形成SC。从早粗线期到晚粗线期,由同源配对调整为非同源配对。另外,在一只雌鸡中,第一次观察到,有些细胞的常染色体能正常配对,而性染色体完全不配对的现象。     

中华鳖精母细胞联会复合体的研究

本文以微铺展技术制备中华鳖精母细胞联会复合体标本,经硝酸银染色后电镜观察,分析了SC组型。并与有丝分裂染色体组型相比较,发现二者有着良好的一致性,而且微小染色体的SC结构和着丝粒清晰,未发现形态上有分化的性染色体。中华鳖SC的研究为其细胞遗传学及性别决定机制提供了重要的依据。 Abstract　Synaptonemal Complexes (SC) in Trionyx sinensis spermatocytes prepared with micro-spreading technique and silver staining was analyzed by electron microscopy. The meiotic SC karyotype was constructed from 10 cells and compared with mitotic chromosome karyotype. There is a good agreement between them. The structure and kinetochores of micro-chromosomes are very distinctive on each SC. There does not exist differential sex chromosome.