Precocious formation of synaptonemal-like polycomplexes and their subsequent fate in female Ascaris lumbricoides var. suum

During meiotic interphase, before leptotene, synaptonemal-like polycomplexes are seen in the cytoplasm of the Ascaris lumbricoides oocytes and in the communal anucleate rachis. In some females short intranuclear synaptonemal complexes are present briefly at that early stage. The number of extranuclear complexes increases just before leptotene, some are attached to the pores of the nuclear membrane. During zygotene most polycomplexes disappear. At late pachytene they reappear in some females but not in others. The morphology, when first seen, is that of disorganized filamentous bodies, later lateral elements appear among the filaments. The dimensions of the lateral elements of the polycomplexes are variable. In the male the distribution of polycomplexes among the rachis, the cell cytoplasm, and at the nuclear envelope is similar to that observed in the female.These observations confirm the precocious occurrence of synaptonemal-like polycomplexes reported by Bogdanov (1977). Ascaris lumbricoides thus, uniquely, appears to manufacture synaptonemal complex-like material in the communal cytoplasm of the germ cells prior to the time that the full complement of synaptonemal complexes appears in the nucleus.     

六种鱼的精母细胞联会复合体的电镜观察

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

Identification of two major components of the lateral elements of synaptonemal complexes of the rat

This paper describes the identification of two major components of the lateral elements of synaptonemal complexes of the rat by immunocytochemical techniques. We prepared monoclonal antibodies against synaptonemal complexes (SCs) by immunization of mice with purified SCs. One of these antibodies, II52F10, reacts with a 30 and a 33 kDa polypeptide, which are major components of purified SCs. Using this antibody, we studied the localization of its antigens light microscopically, by means of the indirect immunoperoxidase technique, as well as ultrastructurally, by means of the immunogold labeling technique. The immunolocalization was carried out on whole-mount preparations of lysed spermatocytes. The antibody reacts with paired as well as unpaired segments of zygotene, pachytene and diplotene SCs. In light microscopic preparations, the attachment plaques, particularly those of late pachytene and diplotene SCs, also appear to react strongly. In electron micrographs the lateral elements in paired as well as unpaired segments could be seen to react. No reaction was observed in the attachment plaques; however, in late pachytene and diplotene SCs the swollen terminal segments of the lateral elements did react with the antibody. Thus, we conclude that a 30 and a 33 kDa polypeptide make part of the lateral elements of synaptonemal complexes of the rat.     

Serial sectioning study of some meiotic stages in Scaptericus borrelli (Grylloidea)

An electron microscope study of the spermatocytes I of Scaptericus borrelli was performed using serial sections and tridimensional reconstruction. The stages examined were: mid-pachytene, early diplotene and the so-called diffuse stage. At mid-pachytene the nucleolus (associated to an autosome) shows a glomerular array formed of supernumerary synaptonemal complexes (extra-complexes, ECs). At early diplotene the autosomes still show remnants of synaptonemal complexes (SC) and the sex chromosome is formed by two substances: one is homogeneous (chromatin-like) the other is granular and dense. A short number of ECs occur in the latter. — At the diffuse stage (lobulated in this paper) the SCs remnants and the ECs have disappeared and the granular material is assembled in a nuclear lobule. This lobule was found associated with one centriole. — A tridimensional model of a diplotene nucleus and models of diplotene bivalents are shown. — The capacity to form repetitive components of the synaptonemal complex type by spermatocytes of the super-Family Grylloidea is discussed.     

Sequential analysis of synaptonemal complexes in the repopulating spermatocytes of Rattus norvegicus after restricting the germ cell population to spermatogonia by gossypol treatment

Synaptonemal complexes of the repopulating spermatocytes of male rats were analyzed day by day using silver-stained surface spread nuclei between 8 and 25 days after restricting the germ cell population to spermatogonia by treatment of gossypol acetic acid at 30 mg/kg body weight/day for 70 days. The method allowed sequential analysis of male meiotic prophase on successive days after the last day of treatment. The leptotene cells appeared on day 11 and were characterized by a network of lateral elements and large nucleolar bodies in a diffuse mass. On day 13 the unpaired lateral elements and short stretches of synaptonemal complexes characteristic for zygotene could be seen. Pachytene nuclei showing 20 autosomal synaptonemal complexes and XY axes appeared on day 15. The diplotene cells were defined on day 22 by the loss of a complete synaptonemal complex set and by the appearance of disjoined lateral elements and persistent segments of synaptonemal complexes.     

Formation of cytoplasmic synaptonemal-like polycomplexes at leptotene and normal synaptonemal complexes at zygotene in Ascaris suum male meiosis

A thread-like (more than 70 cm long) testis of Ascaris suum, when examined under the light and electron microscope, reveals the linear succession of meiotic stages. Beginning from, at least, late leptotene, the spermatocytes are synchronous in their development. Thus within each transverse section of the testis all the spermatocytes are in the same stage. The spermatocytes at each stage of prophase I occupies several (4 to 10) cm of the whole testis length. — At leptotene, synaptonemal-like polycomplexes of lateral and central stacked elements are formed in the cytoplasm of spermatocytes. At late leptotene, the polycomplexes are attached to the external nuclear membrane. The polycomplexes disappear at zygotene. Slightly discernable axial cores are observed in the late leptotene chromosomes. The synaptonemal complexes (SCs) are formed at the zygotene stage, their structure being characteristically tripartite. The SCs disappear from the nuclei at the diffuse stage of prophase I. In other organisms completely developed polycomplexes of stacked lateral and central elements were never found during the presynaptic period of meiosis, although single or two parallel layers of aggregated central regions of SC were found in Neottiella meiocytes at the stage prior to chromosome pairing (Westergaard and von Wettstein, 1970, 1972). — First appearance of the polycomplexes in the cytoplasm insetead of the nucleus is also a novel fact. It is concluded that the polycomplexes at leptotene are formed by a self-assembly of the SC molecular material precociously synthesized in the cytoplasm. Two hypotheses regarding possible function and the further fate for leptotene polycomplexes are discussed.     

Ultrastructural studies of late meiotic prophase nuclei of spermatocytes in Ascaris suum

Post pachytene stages of meiotic prophase in males of Ascaris suum have been analyzed with the electron microscope. No synaptonemal-like polycomplexes (PCs) have been observed in the nucleoplasm or cytoplasm during the period from pachytene to diakinesis. From Serially sectioned diplotene nuclei it was found that the bivalents are located near the periphery of the nuclei, the central part of the nuclei being vacant. Each nucleus contains one nucleolus. Up to 1 m long stretches of unpaired lateral elements (LEs) are found in some of the diplotene bivalents. These LEs are morphologically similar to unpaired LEs in early zygotene nuclei. Partial 3-dimensional reconstruction of two nuclei shows that the bivalents contain some small stretches of synaptonemal complex (SC) up to 1.9 m long. Some bivalents at diakinesis show remnants of SCs. At this stage chromosomes are fibrous, condensed, attached to the nuclear envelope and mostly with a rounded profile in cross section. The synchronous development of the spermatocytes and small bivalents at diplotene in A. suum make this system a good object for the study of localization of SC remnants.     

Evolution of the synaptonemal complex in Helix aspersa spermatocytes

In spermatocytes of Helix aspersa, the structure of the synaptonemal complexes undergoes changes in the course of the pachytene, the lateral elements being transformed into wide bands of lesser density than the chromatin. By using the uranyl-EDTA-lead sequence, which preferentially stains RNA, the lateral elements can be made to appear positive in the early pachytene while the corresponding areas, which become wider and more diffuse, are positive during late pachytene. — Apparently, the lateral elements do not persist in the diplotene and remnants of the central element can occasionally be observed. Using the uranyl-EDTA-lead method reveals some positively stained material surrounding the chromatin, mostly granular in appearance, which is observed in late pachytene and attains its maximum amount during diplotene. Several aspects of these observations are here discussed.     

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)     

The behaviour of chromosomal axes during diplotene in mouse spermatocytes

The fate of the synaptonemal complex and its elements after pachytene has been studied by serial sectioning of diplotene nuclei in mouse spermatocytes. The lateral elements of the synaptonemal complex separate from each other during diplotene, and they form single axes, 300 Å wide, surrounded by chromatin fibrils. The single axes are continuous and end on the nuclear membrane by two different ends: the basal knob and the simple end. The single axes do not cross-over each other, but they remain approached at the convergence regions. In these regions a modified piece of synaptonemal complex is found. This piece changes into a chromatin bridge during diplotene. It has been inferred that the convergence regions represent chiasmata and that the single axes do not represent axial structures of chromatids.     

Abnormalities in synaptonemal complexes in pollen mother cells of a lily hybrid

Two abnormalities, one in lateral elements of synaptonemal complexes the other involving whole complexes, have been studied with the electron microscope in pollen mother cells of the lily hybrid, Lilium aureliensis × L. henryi, which with the light microscope showed almost complete bivalent formation at metaphase. Brief water treatment of pollen mother cells prior to fixation, revealed that the aberrant configurations in lateral elements arose by breakage and subsequent folding of severed fragments up to about 0.8 m long. The abnormality ocurred at recognisable heterologous regions, apparently immediately after pairing. The folded fragments were eliminated from the chromosomes at some time during pachytene. Pseudo pairing was observed after synapsis between either more than one pair of homologues or one pair bent back on themselves, so as to produce polycomplexes. Seemingly, central elements could develop between lateral elements on their outer face under these conditions.     

The development,structure and function of modified synaptonemal complexes in mosquito oocytes

The nucleus of the maturing oocytes expands to a large thin body of 400×140×3 m but the chromosomes remain together in a small sphere, 15 m in diameter. In Aedes aegypti this sphere becomes surrounded by one to several layers of polycomplexes, annulated polycomplexes, and related annulated pseudomembranes. Just prior to egg laying the expanded nucleus disintegrates while the sphere of chromosomes is surrounded by several layers of membranes. In Culex pipiens the elements which normally connect the lateral elements of the synaptonemal complexes become extended so that all bivalents become interconnected by a framework of pseudomembranes. The continuity between the modified synaptonemal complexes and various membranes associated with the karyosphere suggest that a relationship exists, by origin or by specialization, between the synaptic structures and nuclear envelope.     

Spiral cores of synaptonemal complex lateral elements at the diplotene stage in rye include the ASY1 protein

After completing their functioning, synaptonemal complexes (SCs) degrade during the diplotene stage. In the pollen mother cells of rye Secale cereale L., this occurs through the formation of gaps in lateral elements of the SCs and the shortening of fragments of SCs until their complete disappearance. However, when contrasting SCs with silver nitrate solution at a pH 3.5–4.5, these gaps appear to be filled with threads associated with SC lateral elements. As the diplotene stage proceeds and gradual degradation of SC fragments continues, these threads turn into submicroscopic spirals. In this study, we found that the threads and spirals associated with degrading synaptonemal complexes are stained by antibodies to the ASY1 protein of Arabidopsis thaliana lateral elements and thus are degradation products of the lateral elements of SCs.     

Development of the synaptonemal complex and polycomplex formation in three species of grasshoppers

This paper describes the development of the synaptonemal complex in three species of grasshopper: Chorthippus bicolor, Oedipoda coerulescens and Paracinema bicolor. In all three cases the development seems similar. A typical synaptonemal complex is observed during pachytene. Diplotene bivalents show a low density material associated with the chromatin and during first metaphase the beginnings of polycomplex formations are seen. Well organized polycomplexes can be recognized from first anaphase to early spermatids. The elements of the polycomplexes, as well as elements of the synaptonemal complex, show themselves to be positive after preferential staining for ribonucleoproteins. Polycomplexes observed after spreading and positive staining present similar characteristics to those observed after sectioning.This paper is dedicated to the memory of W. Bernhard for his contribution to the knowledge of the cell nucleus     

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.     

Synaptonemal complex and a new type of nuclear polycomplex in three higher plants: Paeonia tenuifolia,Paeonia delavayi,and Tradescantia paludosa

Synaptonemal complex (SC) formation was followed in three species of higher plants: Paeonia tenuifolia, P. delavayi, and Tradescantia paludosa. A desynaptic mutant of the latter species was compared to the wild type. Thickenings of lateral elements and “recombination nodules” were a regular feature of all pachytene SCs. Two types of polycomplexes can be formed in the same species. In diplotene cells of wild-type Tradescantia, polycomplexes of a paracrystalline nature were found in the cytoplasm whereas dense cores were formed in the nuclei. In the desynaptic mutant and in the two Paeonia species, a new type of nuclear polycomplex was observed consisting of the same dense core as seen in the wild type but with a piece of unmodified SC wrapped around it. The number, size, and structure of these “helicoidal polycomplexes” were similar in all nuclei. Their number was equal to the haploid chromosome number of the species: 5 in Paeonia and 6 in Tradescantia.     

Transition from somatic to meiotic pairing and progressional changes of the synaptonemal complex in spermatocytes of Aedes aegypti

Aedes aegypti spermatocytes were reconstructed from electron micrographs. The species has tight somatic pairing of the chromosomes, and there are therefore no classical leptotene and zygotene stages, but rather a gradual transition from somatic pairing to meiotic pairing (= pachytene). The term prepachytene has been used for the transitory stage. The first visible sign of impending meiosis was a reorganization of the chromatin, which resulted in the formation of spaces (synaptic spaces) in the chromatin, about the width of the synaptonemal complexes (SCs). Diffuse material, possibly precursor material for the SC, was present in the spaces. Later short pieces of complex were formed throughout the nucleus. Late prepachytene, pachytene, and diplotene complexes were reconstructed. Each chromosome occupied a separate region of the nucleus. The complexes became progressively shorter from prepachytene (maximum complement length 289 m) to diplotene (175 m). The thickness of the SCs increased from prepachytene to pachytene and probably decreased again during diplotene. At the beginning of diplotene the lateral elements (LEs) separated, and the single LEs became two to three times thicker than the LEs of the SC. The centromeres were at all stages attached to the nuclear membrane, whereas the telomeres were free in the nucleoplasm during pachytene and diplotene. A heterochromatic marker was present on chromosome 1 near the sex determining locus, and a diffuse marker on chromosome 3 near the nucleolus organizer region. After breakdown of the complexes, polycomplexes were present in the nucleus.     

Sequential study of the synaptonemal complex in Syrian hamster (Mesocricetus auratus) and mouse (Mus musculus) oocytes by light and electron microscopy

We describe the behaviour of synaptonemal complexes (SCs) in Syrian hamster and mouse oocytes. InMesocricetus auratus, synaptonemal complexes can be observed from birth up to 7 days of life. In foetuses fromMus musculus, synaptonemal complexes can be observed from the 14th day of gestation up to the first day post-partum, when the cells enter the dictyotene stage. In both species, synaptonemal complexes show, in general, the same morphology described in male cells by light and electron microscopy, with the exception that the axes of the sex bivalent are not identifiable. The leptotene stage can be identified although it is probably of short duration. Only one type of zygotene (zygote ne II of Dietrich and Mulder(Chromosoma 88: 377), 1983) has been observed. In the hamster we also describe a desynaptic diplotene stage previous to the desintegration of the SCs. In oocytes from both species late pairing (or precocious separation) of a single bivalent can be seen in a few cells. Interlocking of some bivalents with delayed pairing of the affected region is rather frequent. Furthermore, hamster oocytes may show heterosynapsis of the telomeres of autosomal bivalents by pachytene.     

Nucleolar structures in chromosome and SC preparations from human oocytes at first meiotic prophase

Summary We describe a comparative study of the behavior of nucleolar structures and their relationship with nucleolar chromosomes and synaptonemal complexes at first meiotic prophase of human oocytes in an attempt to elucidate the nature of this cellular organization and to learn more about maternal nondisjunction. The number of main nucleoli varies along the different stages of prophase I and is usually low. It shows an increase from leptotene to pachytene and a decrease from pachytene to diplotene related to a decrease and an increase of main nucleoli volume, respectively. The methodology employed has enabled us to analyze in detail dark bodies, round bodies, dense bodies, and main nucleoli in chromosome or synaptonemal complex spreads. The relationship between nucleolar chromosomes or synaptonemal complexes and the nucleoli implies the existence, in a very reduced space, of chromosomal regions that contain homologous sequences and that are often unpaired. This situation may facilitate the production of heterologous pairing and chromosomal exchanges between nonhomologous chromosomes and finally result in aneuploidy. THus, the situation explained above together with the differences between the oocyte and spermatocyte NOR cycles could be one of the reasons for the higher incidence of aneuploidies of maternal origin at meiosis I.