The nature of the 1;29 translocation in cattle as revealed by synaptonemal complex analysis using electron microscopy

Synaptonemal complex analyses were carried out by electron microscopy on surface-spread spermatocytes of one normal bull and two bulls that were heterozygous for the so-called 1;29 translocation. The autosomal bivalents of the normal karyotype, which could be arranged by size in a series, demonstrated kinetochores at the terminally located attachment plaques. One autosomal bivalent was clearly larger than the rest and apparently consisted of the long arm of the 1;29 translocation. The 1;29 translocation was the longest autosome in the set and had a kinetochore in a subtelocentric position. Some of the autosome pairs had nucleolus organizer regions in telomeric regions. The X and Y chromosomes, which were not paired at zygotene, demonstrated association in a very short segment at early pachytene; in no cells could a synaptonemal complex be seen between the X and Y. Very often the sex chromosomes were dissociated. At zygotene, a few, usually large, bivalents were unpaired proximally. This always also involved the proximal parts of the arms of the 1;29 translocation and their normal homologs. At early pachytene, the 1;29 trivalent, although to a less extensive degree, was also unpaired in the pericentric region. Configurations in which one chromosome, either 1 or 29, was completely paired with its corresponding arm in the 1;29 translocation chromosome also occurred. When unpaired proximally, the size of chromosome 1 agreed fairly well with the size of its corresponding arm, but the size of chromosome 29 was considerably larger than the corresponding arm of the 1;29 translocation chromosome. During late zygotene and early pachytene, the percent difference between chromosome 29 and its corresponding arm decreased, and at mid and late pachytene there had been a complete synaptic adjustment. The size difference and pairing behavior indicated that a deletion of the kinetochore and the most proximal segment of chromosome 29 had preceded the fusion with chromosome 1 into the 1;29 translocation. The unique structural appearance of the 1;29 translocation chromosome compared to that of other centric fusion translocations in cattle lends support to the theory of a monophyletic origin of the 1;29 translocation. The importance of the pairing behavior observed in governing recombination and chromosome disjunction is briefly discussed.     

Synaptonemal complex analysis in spermatocytes of tilapia, Oreochromis niloticus (Pisces, Cichlidae).

Some adaptations of the synaptonemal complex (SC) whole-mounting technique first used in plants permitted its application to meiotic studies in tilapia, Oreochromis niloticus. Direct observation of the chromosome pairing process and bivalent structure during the meiotic prophase of this fish species by light and electron microscopy permitted the analysis of SCs in autosomes and the possible identification of sex chromosomes. The analysis of SCs in spermatocytes of O. niloticus revealed that all 22 bivalent chromosomes completely paired, except for the occurrence of a size heteromorphism in the terminal region of the largest bivalent associated with the presence of an incompletely paired segment during the synapsis process, which may be the cytological visualization of an XX/XY sex chromosome system in this species.     

Synaptonemal complexes analysis in a bull carrying a 4;8 Robertsonian translocation

Synaptonemal complexes analysis was performed using electron microscopy on surface-spread spermatocytes of a bull heterozygous for the 4;8 Robertsonian translocation. In 19 cells examined, the longest autosomal complex showed kinetochores in a central position whereas the remaining autosomal complexes showed terminal kinetochores. Synapsis in the trivalent appeared complete in all cells, and the trivalents usually showed a CIS configuration. The arm ratio varied from 1.05 to 2.04 with an average of 1.32 +/- 0.43. Out of 47 cells showing X-Y bivalents, 34 showed a small synaptonemal complex at one extremity of the X chromosome, and an unstained gap in the Y chromosome. There was no association between the X-Y bivalent and the trivalent. The absence of association would explain the normal spermatogenesis noted in this bull, in contrast to human and mouse carriers of translocations which show impaired spermatogenesis due to the association between the rearranged chromosomes and the sex vesicle. Further studies involving bulls carrying one or more Robertsonian translocations are needed to determine whether this absence of association is a constant feature in cattle.     

Synaptonemal complex analysis in spermatocytes of diploid and triploid Chinese shrimp Fenneropenaeus chinensis

A modified surface spreading technique for synaptonemal complex (SC) analysis was tested to assess the process of chromosome synapsis in spermatocytes of diploid and induced triploid Fenneropenaeus chinensis. Spermatocytes of diploid shrimp showed typical morphological characteristics of eukaryote SC, with complete synapsis of bivalents. No recognizable bivalent associated with sex chromosomes was observed in spermatocytes of diploid shrimp. However, differences in morphology of SC, including unsynapsed univalents, bivalents, totally paired trivalents with non-homologous synapsis, partner switches and triple synapsis were identified at early pachytene stage of triploid spermatocytes. Triple synapsis was especially common at late pachytene stage in spermatocytes of triploid shrimp. The observed abnormal synapsis behavior of chromosomes in spermatocytes indicated that triploid male shrimp may find it difficult to develop normal haploid sperm.     

XYY spermatogenesis in XO/XY/XYY mosaic mice

The relative frequencies of XYY and XY cells in XO/XY/XYY mosaic mice were compared between somatic cells (bone marrow) and spermatogonia, and between spermatogonia and pachytene or MI spermatocytes. The results indicated there was no selection either for or against XYY spermatogonia. There was, however, a strong selection against XYY spermatocytes during pachytene, with their almost total elimination by the first meiotic metaphase. At pachytene, most XYY cells had trivalent or X univalent/YY bivalent configurations. These findings are contrasted with previous studies of XYY spermatogenesis in mice and are discussed with respect to a model that invokes sex-chromosome univalence as the cause of XYY spermatogenic failure.     

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.     

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.     

Distribution of Atr protein in primary spermatocytes of a mouse chromosomal mutant: a comparison of preparation techniques

In this study, we examined the suitability of a three dimensional preparation technique for studying chromosome behaviour in the first meiotic prophase in the mouse chromosomal mutant T(1;13)H/T(1;13)Wa. To preserve cellular shape, primary spermatocytes were encapsulated in a fibrin clot. Conventionally sedimented prophase nuclei served as controls. Axial elements and lateral synaptonemal complex components were subsequently stained by immunofluorescence and the presence of axial elements at the pachytene stage was highlighted with indirect immunofluorescence against the Atr protein. We compared the distribution of Atr signal in the fibrin-embedded spermatocytes with surface-spread preparations and immunohistochemically stained histological sections of seminiferous tubules. Furthermore, fluorescence in situ hybridisation of the mouse minor satellite DNA was done on fibrin-embedded spermatocytes. The Atr signal is most conspicuous in fibrin-embedded nuclei on unpaired axial elements during pachytene, both for sex chromosomal and for autosomal segments, and expanding from these elements into the surrounding chromatin. Both spread and encapsulated zygotene nuclei with extended axial element formation proved to be positive for Atr. Mid- to late zygotene nuclei were devoid of 3,3′-diaminodibenzene deposition in the histological sections. Highlighting the unpaired axial elements in the small heteromorphic 1¹³H;1¹³Wa bivalent with an Atr signal enabled meiotic analysis of this bivalent to be carried out in a three-dimensional context. Thus, proximity of this bivalent with the sex chromosomes is found more often in three-dimensional preparations than in spread preparations. Furthermore, the development of the Atr signal over the sex chromosomes as pachytene proceeds helps in substaging of this long and heterogeneous meiotic phase, in sedimented but especially in fibrin-encapsulated nuclei. Received: 22 September 1999; in revised form: 20 December 1999 / Accepted: 21 December 1999     

Meiotic configurations in female trisomy 21 foetuses

Summary Analysis of the meiotic configurations formed by the three No 21 chromosomes in oocytes from two trisomy 21 foetuses was undertaken using a spreading technique. Light microscope analysis of the first gave limited resolving power, such that over half the oocytes could not be classified as to presence or absence of trivalent or bivalent plusunivalent. In the second, investigated at the electron microscope level, all 65 cells analysed were informative and precise detail of meiotic pairing in trivalents could be obtained. Two principal forms of trivalent occurred, one in which pairing was initiated at opposite ends of the three No 21's, each initiation point involving only two of the three homologous lateral elements; the other in which pairing was initiated by all three elements at the same end, a triple synaptonemal complex being formed. Only in one oocyte out of the 65 analysed at EM level, however, did triple pairing occur along the entire length of the No 21 trivalent. All others showed splitting into bivalent and univalent at some point along the structure. Unpaired regions within trivalents and all univalents were consistently seen to be thickened and dark staining with silver over the whole period from pachytene to diplotene. This contrasted with the desynapsing lateral elements of previously paired synaptonemal complexes which appeared thin by comparison at diplotene. The significance of the thickening remains, as yet, obscure.     

Non-relationship between nucleolus and sex chromosome system Xyp in Chelymorpha variabilis boheman (Coleoptera: Chrysomelidae)

Nucleolar-organizer region, nucleolus and mode of association of the sex bivalent were analyzed in spermatecytes of Chelymorpha variabilis Boheman. This species (2n=10II+Xy_p) shows the typical sex chromosome system of the group Polyphaga. The results of silver staining techniques showed the nucleolar organizer region localized in a subterminal position of an autosomal bivalent. During meiotic prophase the nucleolus was distinguished with the silver staining and acridine orange fluorescence technique up to diakinesis. The independence of nucleolus and sex bivalent Xy_p during meiosis is demonstrated. The positively silver staining but negatively orange-red material found within the parachute could be involved in the regular co-orientation of both sex chromosomes. After a longer hypotonic treatment, sex bivalents were observed elongated and paired only at one end during the pachytene stage. Along these sex chromosomes, C-bands showed positive blocks located in the pericentromeric and telomeric regions. Heterochromatic association of both sex chromosomes was suggested.     

Human male recombination maps for individual chromosomes

Meiotic recombination is essential for the segregation of chromosomes and the formation of normal haploid gametes, yet we know very little about the meiotic process in humans. We present the first (to our knowledge) recombination maps for every autosome in the human male obtained by new immunofluorescence techniques followed by centromere-specific multicolor fluorescence in situ hybridization in human spermatocytes. The mean frequency of autosomal recombination foci was 49.8+/-4.3, corresponding to a genetic length of 2,490 cM. All autosomal bivalents had at least one recombination focus. In contrast, the XY bivalent had a recombination focus in 73% of nuclei, suggesting that a relatively large proportion of spermatocytes may be at risk for nondisjunction of the XY bivalent or elimination by meiotic arrest. There was a very strong correlation between mean length of the synaptonemal complex (SC) and the number of recombination foci per SC. Each bivalent presented a distinct distribution of recombination foci, but in general, foci were near the distal parts of the chromosome, with repression of foci near the centromere. The position of recombination foci demonstrated positive interference, but, in rare instances, foci were very close to one another.     

Pachytene analysis of a man with a 13q;14q translocation and infertility. Behavior of the trivalent and nonrandom association with the sex vesicle

Pachytene analysis was undertaken in a sterile 13q;14q heterozygous translocation carrier in an attempt to follow the segregational behavior of the trivalent and to evaluate the relationship of Robertsonian translocations in man to the impairment of spermatogenesis. Well-spread bivalents from pachytene nuclei were identified by their chromomere patterns. The trivalent was found always in cis configuration. Silver staining demonstrated the loss of nucleolar organizer regions from the translocated chromosomes. A nonrandom association was found between the trivalent configuration and the sex vesicle in 61% of the pachytene nuclei examined. Such an association has been described before in mice heterozygous for Robertsonian or reciprocal translocations, and may thus represent a general phenomenon. As in mice, this contact was restricted to the centromeric region of the trivalent. A hypothesis relating the association of the trivalent with the sex vesicle to impairment of normal X-chromosome inactivation and subsequent spermatogenic breakdown is discussed. Other chromosomal abnormalities in which sex-vesicle anomalies are associated with male sterility (such as X-or Y-autosomal translocations) are also considered. It is proposed that any process interfering with normal X-chromosome inactivation in pachytene spermatocytes could disturb subsequent meiotic or postmeiotic germ cells development.     

Sex chromosome configurations in pachytene spermatocytes of an XYY mouse

Karyotypic investigation of a phenotypically normal but sterile male mouse showed the presence of an XYY sex chromosome constitution. The synaptic behaviour of the three sex chromosomes was examined in 65 pachytene cells. The sex chromosomes formed a variety of synaptic configurations: an XYY trivalent (40%); an XY bivalent and Y univalent (38.5%); an X univalent and YY bivalent (13.8%); or X, Y, Y univalence (7.7%). There was considerable variation in the extent of synapsis and some of the associations clearly involved nonhomologous pairing. These observations have been compared with previously published information on chromosome configurations at metaphase I from other XYY males.     

Meiosis in four trisomic and one double trisomic males of the newt Pleurodeles waltlii

In the newt Pleurodeles waltlii, meiosis was studied in four trisomic and one double trisomic males. Study of first prophase shows that trivalent frequencies and trivalent configurations are correlated with chromosome length; moreover, trivalent configurations indicate that long chromosomes have multiple points of initiation of synapsis whereas two points might be adequate to secure synapsis of short chromosomes. From the study of metaphase II it appears that the extra chromosomes segregate in half of the spermatocytes II. Some abnormal spermatocytes, resulting from nondisjunction of chromosomes at mitosis or at first division of meiosis, or from precocious division of chromosomes at first division of meiosis were observed. In the male double trisomic meiosis fails at anaphase of second division; this accounts for the sterility of the individual.     

Localised chiasmata due to partial pairing: A 3D reconstruction of synaptonemal complexes in male Stethophyma grossum

The possible role of localised pairing as a mechanism producing localised chiasmata in Stethophyma grossum spermatocytes has been examined ultrastructurally. Nuclei at four successive stages of meiosis from leptotene to pachytene were reconstructed from a series of ultrathin sections and the extent of synapsis as demonstrated by synaptonemal complex (SC) formation was calculated. On the basis of the relative lengths of SCs and condensed chromosomes it was reasoned that only the centromeric ends of the long and medium length bivalents paired, and only one end of these SCs was found attached to the nuclear envelope. Only the three shortest bivalents paired completely, and both their SC ends were attached to the nuclear envelope. Thus pairing was directly related to the distribution of chiasmata. The extent of pairing at different stages suggests that the shortest bivalent paired very quickly, the longer ones progressively slower, and that pairing proceeded zip-like from a point at or very close to the end attached to the nuclear envelope, since all stretches of SC were attached to the envelope, and there were never more than 11 pieces, one for each bivalent. Chromosome decondensation and axial core formation did not occur far in advance of SC formation, and synapsis appeared to be much slower in S. grossum than in other species with non-localised chiasmata, as a larger proportion of the meiotic cysts were in zygotene, compared to Stauroderus scalaris and Locusta migratoria, although this was not quantified.     

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.     

Complete meiotic pairing of crested newt chromosomes.

The longest chromosome (number 1) of Trituturus cristatus carries a heteromorphic segment, a heterozygosity perpetuated by a balanced lethal system. The heteromorphic segment is regarded as achiasmate and has been claimed to be asynaptic. Direct observations of chromosome pairing in spermatocytes and oocytes yield some cases where all homologous chromosomes appear to be completely paired, but the individual bivalents could not be identified as pachytene is not particularly clear in this species. The long arms of bivalent 1 usually remain attached by a terminal chiasma in spermatocytes of T. c. cristatus but the corresponding chiasma is only rarely present in T. c. carnifex spermatocytes. Synaptonemal complexes have been measured in both spermatocytes and oocytes of T. c. cristatus. A karyotype constructed from these measurements matches the main features of somatic and lampbrush chromosome karyotypes, indicating that all chromosomes must be completely paired and proportionately represented as synaptonemal complex. The total length of synaptonemal complex is much the same in spermatocytes and oocytes and is similar to the length in spermatocytes of Xenopus laevis. These two amphibian examples supplement a recent survey of other vertebrate classes to reinforce its conclusion that synaptonemal complex length is not related to genome size in vertebrates.     

随机扩增多态DNA(RAPD)在野蘑菇杂交育种中的应用

野蘑菇 (AgaricusarvensisSchaeffex .Fr.)是蘑菇属 (Agaricus)又一种近年来广泛栽培的食用菌。由于它的菌丝细胞具有多核 ,无锁状联合 ,以及人们对它的繁殖模式和生活史认识的不足 ,给杂交育种工作造成了较多困难。运用随机扩增多态DNA遗传标记技术 ,结合拮抗试验和核相分析 ,对自育的单孢菌株之间的杂交试验进行分析研究。结果表明 :两个相互亲合的同核体菌株被配对培养时 ,交配反应出现 ,并形成异核体的后代。可能野蘑菇具有双重的交配繁殖系统———