Synaptonemal complex proteins

Synaptonemal complexes were isolated from rate spermatocytes for the purpose of biochemical and morphological analysis. Several monoclonal antibodies were elicited against purified synaptonemal complexes to study the composition and assembly of these structures. Four classes of antibodies could be discriminated according to the polypeptides that they recognize on Western blots of purified synaptonemal complexes, namely antibodies recognizing (i) a 190-kDa polypeptide; (ii) a 30- and a 33-kDa polypeptide; (iii) two polypeptides with molecular weights of about 120 kDa; and (iv) polypeptides with molecular weights of 66-55 kDa. The localization of these antigens within spermatocytes was analyzed light microscopically, by means of the immunoperoxidase technique and ultrastructurally, by immunogold labelling of surface-spread spermatocytes. The 66- to 55-kDa polypeptides are not confined to synaptonemal complexes; rather, these polypeptides appear to be chromosomal components. The 190-, 30-, and 33-kDa polypeptides make part of the lateral elements of paired as well as unpaired segments of synaptonemal complexes. The 120-kDa polypeptides were localized on the inner edge of the lateral elements, specifically in paired segments of synaptonemal complexes. The distribution of the 190-, 120-, 30-, and 33-kDa polypeptides within the testis was analyzed by immunofluorescence staining of cryostat sections. All these polypeptides turned out to be specific for nuclei of zygotene up to and including diplotene spermatocytes. Only in some early spermatids could the 190-, 30-, and 33-kDa polypeptides be detected, presumably in remnants of synaptonemal complexes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Shaping meiotic prophase chromosomes: cohesins and synaptonemal complex proteins

Recent progress in elucidating the function of synaptonemal complex (SC) proteins and of cohesins in meiocytes made possible, in particular, through the analysis of mice deficient in SC or cohesin proteins has significantly enriched our understanding of how meiotic chromosome architecture is determined. Cohesins and the SC proteins act together in generating the characteristic axis-loop structure of meiotic chromosomes, their pairing into bivalents, their ability to recombine, and to be properly segregated. This minireview attempts to summarize the current knowledge with a focus on higher eukaryotic systems and to ask questions that ought to be addressed in the future.The synaptonemal complex—50 years     

Immunochemical localization of contractile proteins in mammalian meiotic chromosomes

Summary Smooth muscle heavy myosin and actin have been detected in mouse and rat meiotic chromosomes, by indirect immunofluorescence performed on testis cryostat sections and isolated germ cells. Both contractile proteins are detectable in the nuclei of meiotic cells during the first prophase. The appearance and disappearance time of myosin and actin, however, is not synchronous. While actin is visible in small spots from resting to late diplotene spermatocytes, myosin appears as filaments in the primary spermatocytes from the zygotene to the early stage of diplotene. The number of myosin filaments in the pachytene spermatocytes corresponds to the number of bivalent chromosomes, whereas actin spots constantly outnumber the pairing chromosomes by two units. These immunochemical observations suggest that the two contractile proteins are associated with the synaptonemal complex (SC). Myosin seems to be associated with the central region of the SC, while actin is present in its basal knob which is in connection with the nuclear membrane. The difference in number between myosin filaments and actin spots appears to be related to the peculiar behaviour of the pairing sex chromosomes. The presence of contractile proteins in the nuclei of primary spermatocytes seems to suggest that they might play a role in the process of pairing of homologous chromosomes.     

The Synaptonemal complex component C(2)M regulates meiotic crossing over in Drosophila

BACKGROUND: The synaptonemal complex (SC) is a proteinaceous structure that forms between homologously paired meiotic chromosomes. Previous studies have suggested that the SC is required for meiotic crossing over in Drosophila. However, only one component of this structure, C(3)G, has been identified in Drosophila. RESULTS: Mutations in c(2)M cause a reduced frequency of meiotic crossing over due, in part, to how recombination events are resolved. Cytological evidence suggests that C(2)M is a component of the SC and is required for the assembly of C(3)G (a putative transverse filament of the SC) along the chromosomes. Additionally, C(2)M localizes along the chromosomes in the absence of C(3)G. Despite having a defect in C(3)G localization, c(2)M mutants unexpectedly affect crossing over less severely than a c(3)G mutant. There is virtually no crossing over in a c(3)G mutant, but c(2)M or c(2)M; c(3)G double mutants produce a substantial number of crossovers. The appearance of C(3)G-independent crossovers in c(2)M mutants suggests that C(2)M prevents recombination in the absence of complete SC formation. CONCLUSIONS: We have identified a new Drosophila SC component, C(2)M, that promotes the formation of crossovers. Furthermore, the appearance of C(3)G-independent crossovers in c(2)M mutants suggests a novel role in preventing recombination in the absence of complete SC.     

Whole mount electron microscopy of meiotic chromosomes and the synaptonemal complex

The mechanism by which homologous chromosomes pair and crossover has been a major unsolved problem in genetics. Thin section electron microscopy of the synaptonemal complex has not provided enough details to allow any significant insight into this problem. Whole mount preparations of the testis of mice, quail, crayfish, and frogs provided a striking improvement in visualization of the morphological features of meiotic chromosomes. These studies, when combined with the use of deoxyribonuclease and trypsin allowed the following conclusions. 1. The synaptonemal complex (lateral and central elements with connecting L-C fibers) is composed of protein. 2. Contrary to common speculation the central element is not the pairing surface of homologous chromosomes. 3. The L-C fibers, averaging 75–100 Å in width, extend from the lateral elements and meet to form the central element which is usually composed of four fibers. 4. During leptotene, homologous axial elements, although unpaired for most of their length, attach next to each other at the nuclear membrane. 5. Short segments of the chromatin fibers attach to the lateral elements. These points of attachment are clustered, producing the chromomeres seen by light microscopy. 6. The chromatin fibers extend out from the lateral element as loops. Lampbrush chromosomes are thus not restricted to oogenesis but are common to all meiotic chromosomes.Since the morphological features of the central element of the synaptonemal complex persist despite extensive deoxyribonuclease digestion, pairing is perhaps best visualized as a two-step process consisting of a) chromosomal pairing during which the proteinaceous synaptonemal complex pulls homologous chromosomes into approximate association with each other, and b) molecular pairing, which probably takes place in the area around the synaptonemal complex.Supported by NIH Grants GM-15886 and C-2568, and The Charles and Henrietta Detoy Research Fellowship.     

Synaptonemal complex damage in relation to meiotic chromosome aberrations after exposure of male mice to cyclophosphamide

The genetic implications of induced synaptonemal complex (SC) damage are not known. However, on theoretical grounds, such aberrations could be involved in mechanisms leading to potentially heritable defects. Cyclophosphamide (CP), a chemical reported to cause structural and numerical chromosomal aberrations in the mouse, was used to determine if SC damage observed in meiotic prophase is related to subsequent metaphase chromosomal aberrations. Male mice were injected i.p. with CP. In some instances, mice were also injected simultaneously with tritiated thymidine to label DNA so that cells could be tracked autoradiographically through spermatogenesis. Prophase, primary metaphase (M1), and secondary metaphase (M2) samples were sequentially harvested at appropriate times from the same individual, and nuclei were examined for aberrations. Correlation coefficients between SC and metaphase chromosome aberrations were calculated. The inclusion of tritium labeling increased the number and significance of positive correlations. Positive correlations were found between (1) dose-dependent total SC damage and damage to M1, and to a lesser extent, M2 chromosomes; (2) SC breaks/fragments and M1 chains/rings as well as isochromatid breaks/fragments; (3) SC asynapsis and M1 chromatid breaks/fragments; (4) SC multi-axial configurations and M1 chains/rings as well as isochromatid and chromatid breaks/fragments; and (5) SC multi-axial configurations and M2 hyperploidy. These correlations do not define mechanistic or causal relationships between SC and chromosomal damage. However, taken together with the observation that induced SC damage is many times greater than ensuing metaphase chromosome damage, they substantiate SC analysis as a highly sensitive indicator of potentially heritable effects of this (and presumably other) genotoxic agents.     

Synaptonemal complex karyotype of zebrafish

Meiotic cells of zebrafish have been prepared to show synaptonemal complexes (SCs) by light and electron microscopy. Completely paired SCs from both spermatocytes and oocytes were measured to produce an SC karyotype. The SC karyotype resembles the somatic karyotype of zebrafish and has no recognisable sex bivalent. Measurements of total SC length indicate that SCs grow longer and develop centromeres during pachytene. Oocytes consistently have longer SCs than spermatocytes, presumably correlated with the reported higher recombination frequency in females than in males.     

联会复合体:减数分裂的结构基础

减数分裂是有性生殖生物产生单倍体配子的特殊分裂方式,其第一次分裂(减数分裂I)过程中同源染色体的行为是最突出的特征。在减数分裂I,同源染色体间形成的联会复合体通过促进和调控程序性DNA双链断裂的形成和修复,确保同源染色体正确的识别、配对、重组和分离,从而为减数分裂I的顺利完成提供保障。本综述对联会复合体的组成和功能研究进展进行了回顾,探讨了联会复合体的组装如何影响程序性DNA双链断裂的修复和交叉互换的形成,并总结了与人类生殖障碍相关的联会复合体成分突变,还对该领域未来研究方向进行了展望。     

Giemsa banding of meiotic chromosomes

Applying a Giemsa staining technique to the meiotic chromosomes of Anemone blanda demonstrates that Giemsa bands similar to those seen in the mitotic chromosomes are discernible at all the principal stages of meiosis. The bands are not a product of the Giemsa procedure since they can be seen in unstained preparations using phase-contrast optics as chromocentres in interphase nuclei and as condensed regions in prophase chromosomes. That the bands seem to be permanent features of the nucleus, whether it is dividing or otherwise is an important consideration for understanding their nature and function. Bands and chiasmata do not coincide indicating on the one hand that chiasmata are not responsible for differences in banding patterns and on the other hand that the conservation of bands is an indication that they are either inert regions or specialised regions with considerable adaptive significance. These alternatives can only be resolved by genetical studies of the banding phenomena.     

The meiotic chromosomes of man

Summary Information was obtained on the chromosome number, and the behavior of autosomes as well as of the sex chromosomes in meiosis in human male germ cells derived from 25 Japanese patients, 4 to 79 years in age, who were hospitalized mostly due to epididymitis, prostate cancer, undescended testes or infertility.In 16 out of the 25 specimens, the chromosome numbers, 46 in 2n and 23 in n, were consistently established together with an XY sex-determining mechanism based on spermatogonial and spermatocyte divisions. No reliable counts were obtained from the remaining 9 cases, because of that they provided no cells for precise investigation.The X and Y chromosomes during the leptotene stage were observed as two separate heteropycnotic bodies lying along the inner wall of the nucleus, while at pachytene they formed a sex-vesicle after homologous pairing. At the diplotene, diakinesis and first metaphase the X and the Y appeared as an isopycnotic bivalent showing an end-to-end association, though there were some cells in which they remained as two separate entities free from contact. Evidence was presented that the X and the Y seemed to associate with each other at the distal end of the short arm of each element.One or sometimes two smallest autosomal bivalents tended to show rather precociously a chiasma-terminalization at the first metaphase.The metaphase chromosomes of the second spermatocytes were evident by the haploid number as well as by their widely diverged chromatids with a characteristic spiral configuration.The testicular materials under study contained in most cases polyploid cells with a considerable frequency in spermatogonia as well as in first and second spermatocytes. Giant sperm heads were observed not infrequently, mostly being abnormal in shape. No significant correlation was obtained between the frequency of polyploid cells and the age of patients so far studied.Contribution No. 679 from the Zoological Institute, Faculty of Science, Hokkaido University, Sapporo. — It is our pleasure to dedicate this paper to Professor Dr. Hans Bauer, Max-Planck-Institut für Meeresbiologie, Tübingen, in honor of his sixtieth birthday.     

Synaptonemal complex spreading in Allium

A surface-spreading technique for synaptonemal complexes was applied to triploid Allium sphaerocephalon L. (Liliaceae). In early pachytene two of the three axial elements of each set of three homologues are synapsed, the third is intimately aligned with and accompanies them throughout their whole length. The unsynapsed axis is attached to the synaptonemal complex of the other 2 at up to 50 association sites per trivalent. The distribution of these sites within the trivalents is not even; they are under-represented in the proximal regions. From nought to eight switches (pairing partner exchanges), where the accompanying axis joins in synapsis in exchange for one of the two other strands, occur per trivalent. Very often the telomeres of the aligned axes are attached to their synapsed counterparts by dense spherules, which makes this type of association different from the interstitial ones. Frequently the unsynapsed axes show a double structure along short distances. In late pachytene the intercalary associations are abolished, allowing the unsynapsed axes to engage in various types of non-homologous pairing. Since the association sites involve homologous chromosomes and are less abundant in the pericentric regions (which are usually the last to synapse), it is conceivable that similar structures are responsible for the pre-synaptic alignment of homologues and provide the initiation sites for synaptonemal complex formation in diploids.     

Synaptonemal complex karyotype of Eimeria tenella

In most organisms, biological variability rests on the behaviour of the chromosomes in the meiotic context. Despite the importance of meiosis, very little is known about the meiotic behaviour of the Eimeria chromosomes. The aim of the present study is to describe the standard synaptonemal complex karyotype from Eimeria tenella oocyst spreads by electron microscopy. For that purpose, complete sets of pachytene synaptonemal complexes were obtained and the morphological pachytene karyotype was determined. The authors used a previously reported method that overcomes the difficulty of the extreme resistance of protozoan oocysts to disruption and permits the release of intact meiotic chromosomes. The chromosomes were selected under a light microscope and those selected were stained with phosphotungtic acid and studied by transmission electron microscopy. The authors confirmed 14 chromosomes, which were observed as synaptonemal complexes, and the karyotype was constructed by arranging synaptonemal complexes according to their relative lengths and kinetochore position. Components of the synaptonemal complex, lateral elements, central element, recombination nodules and kinetochore were observed. Measures of the kynetochore, width of the synaptonemal complex, diameter of the recombination nodule and length of the telomeres are given. Minimal and no significant differences were found between measures of chromosomes isolated from different Eimeria tenella strains. To the best of our knowledge, the present investigation for the first time identifies and describes the morphological characteristics of the synaptonemal complex of Eimeria tenella during the meiosis that occurs within the oocysts. In addition, the authors provide evidence of the presence of recombination nodules, suggesting that the recombination process may play an important role in the molecular evolution of this parasite.     

Folded chromosomes in meiotic yeast cells: analysis of early meiotic events.

Analysis of folded chromosomes in cells under standard sporulation conditions shows that the g₀ form of the folded genome is used as the entry into meiosis. Premeiotic DNA replication is initiated from the g₀ structure. In contrast, mitotic DNA replication is preceded by a characteristic pre-replicative form, g₁. Nonetheless, the mitotic and meiotic replication structures are indistinguishable by sedimentation. Preliminary evidence also suggests that the meiotic equivalent of the mitotic post-replicative structure, g₂, is absent. In strains homozygous for the mating type locus, aa and αα, meiotic replicating structures are not detected, and the folded chromosomes remain in a non-cycling form. However, this non-cycling form is distinguishable from the g₀ form of aα. cells.     

DNA-binding proteins related to the dosage of specific yeast chromosomes

A comparison has been made of the $\text{in vitro}$ DNA-binding proteins of specific aneuploid and isogenic euploid cells of $\text{Saccharomyces cerevisiae}$ by DNA-cellulose chromatography. We have been able to detect changes in the level of a small fraction of the yeast DNA-binding proteins which can be related to the dosage of specific yeast chromosomes. At least four proteins show a dosage related to the cellular level of chromosome I and at least one protein shows a dosage related to the level of chromosome VI.     

Synaptonemal complex antigen location and conservation

The axial cores of chromosomes in the meiotic prophase nuclei of most sexually reproducing organisms play a pivotal role in the arrangement of chromatin, in the synapsis of homologous chromosomes, in the process of genetic recombination, and in the disjunction of chromosomes. We report an immunogold analysis of the axial cores and the synaptonemal complexes (SC) using two mouse monoclonal antibodies raised against isolated rat SCs. In Western blots of purified SCs, antibody II52F10 recognizes a 30- and a 33-kD peptide (Heyting, C., P. B. Moens, W. van Raamsdonk, A. J. J. Dietrich, A. C. G. Vink, and E. J. W. Redeker, 1987, Eur. J. Cell Biol., 43: 148-154). In spreads of rat spermatocyte nuclei it produces gold grains over the cores of autosomal and sex chromosomes. The cores label lightly during the chromosome pairing stage (zygotene) of early meiotic prophase and they become more intensely labeled when they are parallel aligned as the lateral elements of the SC during pachytene (55 grains/micron SC). Statistical analysis of electronically recorded gold grain positions shows that the two means of the bimodal gold grain distribution coincide with the centers of the lateral elements. At diplotene, when the cores separate, the antigen is still detected along the length of the core and the enlarged ends are heavily labeled. Shadow-cast SC preparations show that recombination nodules are not labeled. The continued presence suggests that the antigens serve a continuing function in the cores, such as chromatin binding, and/or structural integrity. Antibody III15B8, which does not recognize the 30- and 33-kD peptides, produces gold grains predominantly between the lateral elements. The grain distribution is bimodal with the mean of each peak just inside the pairing face of the lateral element. The antigen is present where and while the cores of the homologous chromosomes are paired. From the location and the timing, it is assumed that the antigen recognized by III15B8 functions in chromosome pairing at meiotic prophase. The two anti-rat SC antibodies label rat and mouse SCs but not rabbit or dog SCs. A positive control using human CREST (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia) anti-centromere serum gives equivalent labeling of SC centromeres in the rat, mouse, rabbit, and dog. It is concluded that the SC antigens recognized by II52F10 and III15B8 are not widely conserved. The two antibodies do not bind to cellular or nuclear components of somatic cells.(ABSTRACT TRUNCATED AT 400 WORDS)