Dependence on genic balance for synaptonemal complex formation in Drosophila melanogaster

Electron microscopic examination of gonads of Drosophila melanogaster with different genotypes, including a metafemale 3X;2A and an intersex XXY;3A have revealed that the formation of synaptonemal complexes is controlled by the genic balance, i.e., the ratio of X chromosomes to autosomes. The Y chromosome is not involved in the genetic control of the formation of precursors of the central element of synaptonemal complexes in males, nor does it disturb their formation in XXY females. Hyperploidy for sections 1-3A and 18A-20 of the X chromosome does not lead to the appearance of synaptonemal complexes in males and does not interfere with their formation in females. Females hyperploid for extensive regions of the X chromosome (sections 1-11A, 11A-20, and 8C-20) are fertile and show apparently normal formation of synaptonemal complexes. Hyperploidy for sections 8C-11A of the X results in a sharp decrease in the viability of females, in abnormal differentiation of ovary cells, and in the lack of synaptonemal complexes. These data suggest a possible important role for the sections 8C-11A in the genic balance controlling the formation of synaptonemal complexes in D. melanogaster. The lack of synaptonemal complexes in hypoploid females may be the result of abnormal cell differentiation in gonads.     

Chromosomal pairing in deer mice heterozygous for the presence of heterochromatic short arms

The pattern of chromosomal pairing was analyzed in male deer mice (Peromyscus maniculatus and Peromyscus sitkensis) heterozygous for the presence of heterochromatic short arms. G- and C-banding of somatic metaphases indicated that the presence of heterochromatic short arms increased the length of chromosome 4 by 15% in P. sitkensis and that of chromosome 8 by 9% in P. maniculatus. Analysis of silver-stained late zygotene and early pachytene nuclei revealed a low frequency of unequal axial lengths in the synaptonemal complexes corresponding to the heteromorphic bivalents. All mid- and late pachytene nuclei, however, exhibited fully paired synaptonemal complexes with equalized axial lengths. These observations suggest the existence of an adjustment mechanism which functions to equalize the lengths of the two axes of the heteromorphic synaptonemal complex.     

Stage-specific damage to synaptonemal complexes and metaphase chromosomes induced by X rays in male mouse germ cells

Synaptonemal complexes reveal mutagen-induced effects in germ cell meiotic chromosomes. This study was aimed at characterizing relationships between damage to synaptonemal complexes and metaphase I chromosomes following radiation exposure at various stages of spermatogenesis. Male mice were irradiated with doses of 0, 2, or 4 Gy, and spermatocytes were harvested at times consistent with earlier exposures as spermatogonial stem cells, preleptotene cells (premeiotic DNA synthesis), or meiotic prophase cells. After stem-cell exposure, twice as many rearrangements were observed in synaptonemal complexes as in metaphase I chromosomes. Irradiation during premeiotic DNA synthesis resulted in dose-related increases in synaptonemal complex breakage and rearrangements (including novel forms) and in metaphase chromosomal aberrations. Following prophase exposure, various types and levels of damage to synaptonemal complexes and metaphase chromosomes were observed. Irradiation of zygotene cells led to high frequencies of chromosome multivalents in metaphase I without a correspondingly high level of damage in preceding prophase synaptonemal complexes. Thus irradiation of premeiotic and meiotic cells results in variable relationships between damage to synaptonemal complexes and metaphase chromosomes. Interpretations of these relationships are based upon what is known about both radiation clastogenesis and the structural/temporal relationships between synaptonemal complexes at prophase and chromosomes at metaphase I of meiosis.     

SYNAPTONEMAL COMPLEXES IN RED ALGAE1,2

The presence of synaptonemal complexes in the nuclei of young tetraspore mother cells is described for the first time in the red algae. Synaptonemal complexes were found in Janczewskia gardneri Setchell, Levringiella gardneri Setchell, Gonimophyllum skottsbergii Setchell, and Polycoryne gardneri Setchell. The synaptonemal complexes consist of 2 lateral, dark-staining elements from which small fibrils extend to form a less densely staining central element. With minor variations, the dimensions and structure of these synaptonemal complexes correspond to those found in other organisms.     

Meiosis in Bombyx mori females.

Crossing over is absent in oocytes of the silkworm, Bombyx mori. Synaptonemal complexes are present during pachytene between the paired chromosomes. At leptotene, lateral components of the synaptonemal complex are attached in a bouquet to a limited region of the nuclear envelope. Before completion of lateral components, synaptonemal complex formation begins at the nuclear envelope. With synaptonemal complex formation proceeding from both ends bivalents occasionally become interlocked. After pairing is completed, the bouquet arrangement is dissolved possibly as a result of a flow of the inner membrane of the nuclear envelope thereby separating the telomeres. After the telomeres are released from the nuclear envelope, material is deposited onto the lateral components of the synaptonemal complex. The modified synaptonemal complexes are retained by the bivalents until metaphase I. It is suggested that these modified synaptonemal complexes substitute for chiasmata in order to ensure regular disjunction of homologous chromosomes in the absence of crossing over.     

Fine structure of chromosome pairing in ten Ascomycetes: meiotic and premeiotic (mitotic) synaptonemal complexes

The meiotic prophase of ten Ascomycetes was studied by both light and electron microscopy. All ten species show a Neurospora type of meiosis. Two Podospora have no distinct synaptonemal complexes but synaptic structures in their nucleolus. The synaptonemal complexes of the eight other species are well differenciated and have uniform dimensions. The banded pattern of the lateral components seems to be a characteristic of Ascomycetes. The most striking observation is the specificity of this banding which can be used as a marker in interspecific crosses. Ascophanus carneus shows two kinds of synaptonemal complexes: normally between the pachytene bivalents and unusually in the ascogenous hyphae mitoses before meiosis and even before caryogamy.     

Spermatogenesis in Bombyx mori. II. The ultrastructure of synapsed bivalents

In Bombyx mori the male is the homogametic sex, crossing over occurs only in males, and chiasmata are observed in spermatocytes, but not in oocyte nuclei. If the assembly of synaptonemal complexes is an essential prerequisite for genetic crossing over and chiasmata formation, then the nuclei of Bombyx spermatocytes should contain synaptonemal complexes. Synaptonemal complexes were found in spermatocytes from young four instar larvae. The structure of meiotic bivalents is described using micrographs taken with 100 and 1000 KV electron microscopes. These data together with that from the literature are used to construct a three-dimensional model of the synaptonemal complex and to suggest its method of origin and its function during crossing over.     

The Effect of Bovine Serum Albumin and Cytohelicase on Surface-Spread Synaptonemal Complexes of Rye (Secale Cereale)

Synaptonemal complexes of rye meiocytes were spread on plastic coated slides for electron microscopic observation. Two proteins generally used in synaptonemal complex spreading techniques, bovine serum albumin and cytobelicase, were applied separately or in combination in an isotonic protoplast medium at concentrations of 0.1-5%. At high concentrations these proteins proved to enhance notably the ultimate number of cells with synaptonemal complexes in the preparations. Also under this condition, centromere structures became stainable with silver nitrate in both the synaptonemal complexes of pollen mother cells and in interphase nuclei of other cell types. Since the true action of cytohelicase under appropriate spreading conditions was uncertain, the putative enzymatic digestion of cell walls was determined in a series of experiments using the fluorochrome calcofluor white as a stain of callose walls. Obvious breakdown of the cell walls was not observed before 8 min of treatment under standard conditions. This made it plausible that the prime effect of cytohelicase is that of a nonspecific protein interacting with the chromatin and improving the adhesion of synaptonemal complexes to the hydrophobic plastic film. The differential staining of the centromere structures in the presence of bovine serum albumin and cytohelicase probably reflects a reduced spreading of these structures due to preferential binding between these proteins and centromeric proteins.     

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.     

Distribution of MLH1 foci on the synaptonemal complexes of chicken oocytes

The frequency and distribution of the mismatch repair protein MLH1 was analyzed on synaptonemal complex spreads of chicken oocytes using indirect immunofluorescence. MLH1 foci appeared in late zygotene and their number remains constant throughout pachytene. The average number of foci on autosomal synaptonemal complexes (65.02 +/- 4.02) is in agreement with the number of chiasmata estimated from lampbrush chromosomes. The distribution of foci along the synaptonemal complexes is shown to be nonrandom and nonuniform in terms of the distances between them. It is concluded that MLH1 foci are good markers of crossing over in bird (chicken) meiocytes.     

The synaptonemal complex and the spindle plaque during meiosis in yeast

Meiosis in Saccharomyces cerevisiae proceeds principally in the same manner as in other Ascomycetes. Leptotene is characterized by unpaired lateral components and pachytene by the presence of extensive synaptonemal complexes. The synaptonemal complex has the same dimensions and is similar in structure to those described for other organisms. Chromosome counts can now be made by reconstructing the synaptonemal complexes. Diplotene nuclei consistently contain a single polycomplex. The behaviour, doubling and the fine structure of the spindle plaque provide additional markers for the different stages of meiosis.     

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.     

Coelomomyces psorophorae vartasmaniensis Couch+Laird (1988) (Coelomomycetaeceae: Blastocladiales), a fungal pathogen of the mosquitoAedes australis

The presence of synaptonemal complexes were checked in dividing chromosomes as evidence for meiotic division in germinating sporangia. Continuous urografin gradients were used to separate out the various phases of germinating sporangia, the nuclei were removed and embedded for ultrastructural studies. Meiotic inhibitors were applied to germinating sporangia to retard meiotic division to highlight the synaptonemal complexes. At an early phase of sporangial differentiation dividing nuclei developed with synaptonemal complexes. Meiotic inhibitors and stimulators may be used to control sporangial germination for an induction of a high meiospore count. This may be of crucial importance in the utilization of Coelomomyces spp. as a biological control agent of mosquito species.     

The effect of bovine serum albumin and cytohelicase on surface-spread synaptonemal complexes of rye (Secale cereale)

Synaptonemal complexes of rye meiocytes were spread on plastic coated slides for electron microscopic observation. Two proteins generally used in synaptonemal complex spreading techniques, bovine serum albumin and cytohelicase, were applied separately or in combination in an isotonic protoplast medium at concentrations of 0.1-5%. At high concentrations these proteins proved to enhance notably the ultimate number of cells with synaptonemal complexes in the preparations. Also under this condition, centromere structures became stainable with silver nitrate in both the synaptonemal complexes of pollen mother cells and in interphase nuclei of other cell types. Since the true action of cytohelicase under appropriate spreading conditions was uncertain, the putative enzymatic digestion of cell walls was determined in a series of experiments using the fluorochrome calcofluor white as a stain of callose walls. Obvious breakdown of the cell walls was not observed before 8 min of treatment under standard conditions. This made it plausible that the prime effect of cytohelicase is that of a nonspecific protein interacting with the chromatin and improving the adhesion of synaptonemal complexes to the hydrophobic plastic film. The differential staining of the centromere structures in the presence of bovine serum albumin and cytohelicase probably reflects a reduced spreading of these structures due to preferential binding between these proteins and centromeric proteins.     

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)     

Synaptonemal Complexes from Dnase-Treated Rat Pachytene Chromosomes Contain (Gt)(n) and Line/Sine Sequences

Purified chromosome cores (synaptonemal complexes) of rat pachytene chromosomes, from which the chromatin is removed by extensive DNase II digestion, retain a residual class of DNA, presumably the bases of chromatin loops. This synaptonemal complex-associated DNA, isolated by proteinase digestion and phenol extraction of purified DNase-treated synaptonemal complexes, and cloned in plasmid vector pEMBL18, has a length distribution of 50-500 bp. From a library of these fragments, 21 fragments were sequenced. Present in this sample are short 40-200-bp segments with >80% identity to ``long'''' and ``short'''' interspersed repeated elements (LINE/SINEs), an excess of GT/CA tandem repeats and a number of unidentified sequences. The LINE/SINE segments may play a role in homology vs. nonhomology recognition during meiosis and the alternating purine-pyrimidine sequences have been implicated in genetic recombination. Their enrichment in synaptonemal complexes may be related to the synapsis and recombination functions of meiosis.     

Alteration of meiotic chromosomal pairing and synaptonemal complexes by cycloheximide

When cells are exposed to cycloheximide during the synaptic period of meiotic prophase, the structure of the synaptonemal complex is markedly altered. The bulk of the lateral component is removed. When lily zygotene microsporocytes are subsequently transferred into a culture medium free from cycloheximide, normal synaptonemal complexes are again seen. Modification of the structure of the synaptonemal complex by treatment with cycloheximide for 4 days has little permanent effect on meiosis except at late zygonema or early pachynema. Treatment at this time produces meiocytes in which no synaptonemal complexes reform. When these cells proceed into diplotene and diakinesis they are devoid of chiasmatic chromosomes. The data suggest that the synaptonemal complex is essential if chiasmata are to be formed, and that a unique period exists when the formation can be interrupted.This work was supported by grants from the National Science Foundation (GB 5173X and GB 6476) and the National Institutes of Health (GM 16882).     

Synaptonemal complexes in vaccinated mice

Synaptonemal complexes of spermatocytes I obtained from C57BL/6j male mice treated with inactivated bacterial vaccines were spread over the hypotonic phase and then were investigated using light microscope. The slides of synaptonemal complexes of mice treated with cyclophosphamide were used as positive control. It is shown possible in principle to reveal synaptonemal complex abnormalities by means of light microscopy. These abnormalities were not more frequent in vaccinated animals than in intact ones. Cyclophosphamide at doses of 100-200 mg/kg induced synaptonemal complex damage practically in 100% of cells 96 hours after the injection.     

A Simple Method of Staining the Synaptonemal Complex with Coomassie Brilliant Blue for Light Microscopy

A simple technique for staining synaptonemal complexes with Coomassie brilliant blue for light microscopy has been described. The testis cells were exposed to prolonged hypotonic treatment and dropped on Formvar-coated slides. Following fixation with paraformaldehyde the slides were stained with Coomassie brilliant blue for 15 min to 1 h at room temperature and rinsed in distilled water. For its simplicity and rapidity this technique may serve as an effective alternative to silver staining for light microscopic observation of synaptonemal complexes.     

SC-karyotyping on electronic microscopy (EM) analysis of synaptonemal complexes (SC) spermocytes of silver fox (Vulpes fulvus)

We present an electronic microscopy (EM) analysis of synaptonemal complexes (SC) spermatocytes of male silver fox Vulpes fulvus at the pachytene stage. The SC-karyotypes of pachetene cells were made and described. Knowledge of normal SC-karyotype is necessary to reveal synaptic abnormalities of autosome and sex bivalents during the pachytene. It was indicated that EM analysis of SC-spermatocytes—the study of synaptonemal complexes in agricultural animals—is a very good instrument for comparative analysis of normal SC-karyotype of foxes that carry chromosomal abnormalities and in fur-bearing animals as well.