Occurrence of synaptonemal complexes and recombination nodules in a meiotic race of Meloidogyne hapla and their absence in a mitotic race

The early prophase of the first maturation division of oocytes is compared in two races of Meloidogyne hapla Chitwood. Light microscopic analysis suggested that chromatin of both Race A (meiotic parthenogenetic; n=17) and Race B (mitotic parthenogenetic; 3n=45) behaves similarly during early stages of maturation of oocytes (Triantaphyllou, 1966). Electron microscopic analysis shows that synaptonemal complexes and recombination nodules are observed at pachytene in Race A, but are absent in corresponding oocytes of Race B. Cylindrical granular complexes present in the cytoplasm at prepachytene stages are intimately associated with the nucleus at pachytene in Race A, but are absent in Race B.Paper number 5599 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, North Carolina 27650     

Karyotype analysis of Meloidogyne hapla by 3-D reconstruction of synaptonemal complexes from electron microscopy of serial sections

Meloidogyne hapla Race A (meiotic, n=17) females have 17 synaptonemal complexes (SC). The karyotype length is constant throughout pachytene, although nuclear volume increases as pachytene progresses. Each SC has at least one region in which two pairs of lateral elements run parallel to each other for a distance of 1–2 m, thus forming a double SC (dSC). Decondensed chromatin regions (DCR) occur along some SCs and represent 5% of the length of the karyotype. The DCRs may be the location of the sex-determining chromatin. Spermatocytes from males of the same meiotic parthenogenetic race (A) have SCs and cylindrical granular complexes (CGC) while males and females from a mitotic (3n=45) parthenogenetic race (B) lack such structures. The CGCs may contribute precursors necessary for the formation of SCs.     

Spreading the synaptonemal complex of Neurospora crassa

A protocol was developed to spread the synaptonemal complex (SC) of the fungus Neurospora crassa. It involves direct mechanical breakage of meiotic cells before spreading. This technique makes it possible to examine the SC of the same nucleus with both light and electron microscopy. This protocol is potentially applicable for other Pyrenomycetes. The SCs were examined at zygotene, pachytene and diplotene. The central elements and the recombination nodules (RN) were well revealed by silver staining. Ten to 13 RNs were counted at pachytene. The total genomic SC length varied with the stage. This whole mount electron microscopy of the SC is particularly useful for studying chromosomal rearrangements.     

Three-Dimensional Ultrastructural Karyotype Analysis from the Meiotic Parthenogenetic Nematode Heterodera betulae

Meiotic chromosome structure and function are described in the plant-parasitic nematode Heterodera betulae. Twelve synaptonemal complexes (SCs) were reconstructed from pachytene nuclei; therefore, n=12 is predicted for this species. Morphologically distinct sex chromosomes were not observed. Only one end of the SC is attached to the nuclear envelope, and there is no bouquet arrangement at pachytene. The structure of the SC in this meiotic parthenogenetic nematode was different than in other nematodes that reproduce via amphimixis; a striated central element with transverse filaments was not observed. Multiple SCs, or polycomplexes, were present in the nucleus. Recombination nodules were not observed. The centrioles were comprised of nine doublet microtubules connected by a ring, which is a distinct modification from the typical nine triplet microtubules without any interconnecting structure.     

Enzymatic Relationships and Evolution in the Genus Meloidogyne (Nematoda: Tylenchida)

Thirty populations of Meloidogyne of diverse geographic origin representing 10 nominal species and various reproductive, cytological, and physiological forms known to exist in the genus were examined to determine their enzymatic relationships. The 184 bands resolved in the study of 27 enzymes were considered as independent characters. Pair-wise comparisons of populations were performed in all possible combinations to estimate the enzymatic distances (ED) and coefficients of similarity (S). A phylogenetic tree was constructed. The apomictic species M. arenaria, M. microcephala, M. javanica, and M. incognita shared a common lineage. M. arenaria was highly polytypic, whereas conspecific populations of M. javanica and M. incognita were largely monomorphic. The mitotic and meiotic forms of M. hapla were very similar (S = 0.93), suggesting that the apomictic race B evolved only recently from the meiotic race A. The five remaining meiotic species (M. chitwoodi, M. graminicola, M. graminis, M. microtyla, and M. naasi - each represented by a single population) were not closely related to each other or to the mitotic species.     

Unusual and Strongly Structured Sequence Variation in a Complex Satellite DNA Family from the Nematode Meloidogyne chitwoodi

An AluI satellite DNA family has been isolated in the genome of the root-knot nematode Meloidogyne chitwoodi. This repeated sequence was shown to be present at approximately 11,400 copies per haploid genome, and represents about 3.5% of the total genomic DNA. Nineteen monomers were cloned and sequenced. Their length ranged from 142 to 180 bp, and their A + T content was high (from 65.7 to 79.1%), with frequent runs of As and Ts. An unexpected heterogeneity in primary structure was observed between monomers, and multiple alignment analysis showed that the 19 repeats could be unambiguously clustered in six subfamilies. A consensus sequence has been deduced for each subfamily, within which the number of positions conserved is very high, ranging from 86.7% to 98.6%. Even though blocks of conserved regions could be observed, multiple alignment of the six consensus sequences did not enable the establishment of a general unambiguous consensus sequence. Screening of the six consensus sequences for evidence of internal repeated subunits revealed a 6-bp motif (AAATTT), present in both direct and inverted orientation. This motif was found up to nine times in the consensus sequences, also with the occurrence of degenerated subrepeats. Along with the meiotic parthenogenetic mode of reproduction of this nematode, such structural features may argue for the evolution of this satellite DNA family either (1) from a common ancestral sequence by amplification followed by mechanisms of sequence divergence, or (2) through independent mutations of the ancestral sequence in isolated amphimictic nematode populations and subsequent hybridization events. Overall, our results suggest the ancient origin of this satellite DNA family, and may reflect for M. chitwoodi a phylogenetic position close to the ancestral amphimictic forms of root-knot nematodes. Received: 23 April 1997 / Accepted: 9 July 1997     

Separation of roles of Zip1 in meiosis revealed in heterozygous mutants of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Synapsis of homologs during meiotic prophase I is associated with a protein complex built along the bivalents—the synaptonemal complex (SC). Mutations in the SC-component gene ZIP1 diminish SC formation, leading to reduced recombination levels and low spore viability. Here we show that in SK1 strains heterozygous for a deletion of ZIP1 in certain regions meiotic interference are impaired with no decrease in recombination levels. The extent of synapsis is over all reduced and NDJ levels of a large endogenous chromosome and of artificial chromosomes (YACs) rise to twice the level of wild type strains. A substantial proportion of mis-segregating YACs had undergone crossing over. This demonstrates that different functions of Zip1 display differential sensitivities to changes in expression levels.     

The synaptonemal complex of Meloidogyne nataliei and its relationship to that of other Meloidogyne species

The four synaptonemal complexes (SC) in Meloidogyne nataliei (n=4) have normal, tripartite morphology, although the total width of the SC is only 50 nm. Each SC is attached at both ends to the nuclear envelope and there is no bouquet formation at pachytene. Pairing of homologues is regular but not complete, as there is one region on each bivalent where the SC is not formed. This region may be the fusion point of telomeres of nonhomologous chromosomes, since it is assumed that M. nataliei has been derived from an ancestral strain (n=8) via chromosomal fusions. In each pachytene nucleus there is a nonmembranous, vacuole-like structure of unknown function located in the nucleoplasm adjacent to the nucleolus, and of approximately the same volume as the nucleolus. The fact the SC structure of M. nataliei is strikingly different from that of most Meloidogyne species suggest that M. nataliei may not belong to the same phyletic group as the genus Meloidogyne.     

Meiosis in gray voles of the subgenus <Emphasis Type="Italic">Microtus</Emphasis> (Rodentia,Arvicolinae) and in their hybrids

The results of light and electron microscopic (EM) studies of meiosis in Microtus arvalis males of the karyoform “arvalis” (2n = 46, NFa = 80), in hybrids between the chromosomal forms arvalis and obscurus (2n = 46, NFa = 68), in M. rossiaemeridionalis voles (2n = 54, NFa = 54), and in a hybrid between the species M. rossiaemeridionalis and kermanensis (2n = 54, NFa = 54) are presented. SC (synaptonemal complex) karyotypes of the parental forms and the hybrids were constructed on the basis of measurements of the length of autosomal SCs revealed by the EM analysis in spermatocytes at the stage of middle pachytene. The SC karyotypes of M. arvalis and the hybrids ♀ obscurus × ♂ arvalis consist of 22 synaptonemal complexes of autosomal bivalents and the axial elements of the synaptonemal complexes of the sex chromosomes X and Y. The SC karyotypes of M. rossiaemeridionalis and the hybrid M. rossiaemeridionalis × M. kermanensis consist of 26 synaptonemal complexes of autosomal bivalents and a sex bivalent; they differ only in the length of the Y chromosome axis (Y chromosome in the hybrid was inherited from M. kermanensis). Asynaptic configurations of the autosomal SCs were not observed in the hybrids. The SC axial elements of the X and Y chromosomes in the parental forms and in the hybrids were located close to each other throughout pachytene, but they did not form a synaptic region. The normal synapsis in sterile hybrids (M. rossiaemeridionalis × M. kermanensis) and the behavior of the sex chromosomes in meiosis in fertile and sterile hybrids are discussed in the context of specific features of meiosis and reproductive isolation.     

Meiosis in Mesostoma Ehrenbergii Ehrenbergii. IV. Recombination Nodules in Spermatocytes and a Test of the Correspondence of Late Recombination Nodules and Chiasmata

Male meiosis in Mesostoma ehrenbergii ehrenbergii (2x = 10) is characterized by extreme restriction of chiasma formation; 3 pairs of chromosomes form bivalents at metaphase I which are associated by single very distally localized chiasma, while two pairs of chromosomes remain as unpaired univalents. Electron microscopical three-dimensional reconstruction analysis of serial sections has been applied to 20 pachytene spermatocyte nuclei. In each nucleus three short stretches of synaptonemal complex (SC) were found, confined to a localized branched lobe of the nucleus, confirming the findings of an earlier study. The majority of reconstructed nuclei show that each of the three SC segments has a single prominent recombination nodule ("late" RN) associated with it. Late RNs in this system therefore show an excellent correspondence with metaphase I chiasmata, in contrast to a previous report. M.e. ehrenbergii is therefore not an exception to the hypothesis that meiotic exchange requires a functional late RN. A few nuclei had two, one or no RNs; these presumably represent nuclei that are not at the stage of maximum RN presence. Although M. e. ehrenbergii shows pronounced chiasma localization at the light microscope level, at the ultrastructural level RNs are widely distributed along the 5-10 microns of SC formed in each bivalent, indicating that genetic exchange are not restricted to particular localized sites but occur at a large number of DNA sequence.     

The synaptonemal complexes of Caenorhabditis elegans

Normal synaptonemal complexes (SCs), consisting of two lateral elements and a central element, are present in wild-type, him-4 and him-8 mutant strains in both hermaphrodites and males of Caenorhabditis elegans. Thus, the increase in rate of nondisjunction in the him mutants is not related to aberrant SC morphology. The wild-type hermaphrodite has six SCs, as determined from 3-D reconstruction analysis of serial sections from electron microscopy. Thus, n = 6 and this confirms early reports based on cytological studies with the light microscope. Only one end of the SC is attached to the nuclear envelope while the other end is free in the nucleoplasm and there is no apparent bouquet formation. Either end of the SC can attach to the nuclear envelope. The pairing behavior of the XX bivalent is normal and occurs synchronously with the autosomes. Electron dense bodies, or knobs, are associated with the SC via the central element and displace the chromatin for a distance of 200 nm. Each pachytene nucleus of the wild-type hermaphrodite has six such structures that are randomly dispersed along the bivalents such that some SCs have one or two knobs while others have none. Their function is unknown.     

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

In most organisms the synaptonemal complex (SC) connects paired homologs along their entire length during much of meiotic prophase. To better understand the structure of the SC, we aim to identify its components and to determine how each of these components contributes to SC function. Here, we report the identification of a novel SC component in Drosophila melanogaster female oocytes, which we have named Corolla. Using structured illumination microscopy, we demonstrate that Corolla is a component of the central region of the SC. Consistent with its localization, we show by yeast two-hybrid analysis that Corolla strongly interacts with Cona, a central element protein, demonstrating the first direct interaction between two inner-synaptonemal complex proteins in Drosophila. These observations help provide a more complete model of SC structure and function in Drosophila females.     

A comparative cytological investigation of the reproductive cycle of an amphimictic diploid and a parthenogenetic triploid form of Lumbricillus lineatus (O.F.M.) (Oligochaeta,Enchytraeidae)

Summary The cytology of a diploid amphimictic (2x=26) and a triploid parthenogenetic (3x=39) cytotype of Lumbricillus lineatus has been described. The triploid type never produces mature sperm, but it must be fertilized by sperm from the diploid cytotype in order to produce viable eggs. However, the sperm does not enter the egg, but eggs which have not been activated by sperm die at an early stage. Oogenesis in the triploid cytotype, which results ultimately in the restoration of the somatic chromosome number, follows a pattern which has not been described in other parthenogenetic organisms. The first meiotic division is asynaptic and no typical metaphase plate is formed; the univalents are distributed to opposite poles without undergoing an equational division. The first division is therefore numerically approximately reductional. The nuclei are at anaphase when the eggs leave the animal, whereas the nuclei of the diploid amphimictic cytotype are in MI when the eggs are laid. Following activation by spermatozoa, the first anaphase spindle is much elongated, and becomes V-shaped. The first anaphase spindle persists and no second anaphase spindle is formed. At the second division the chromosomes divide mitotically. The resulting four chromosome groups fuse two by two in such a way that two nuclei are formed each with the full somatic (triploid) chromosome set (cf. Figs. 14–16).The different steps involved in the evolution of this mechanism are discussed in relation to the evolution of the triploid cytotype.     

One class of mutants with disturbed centromere cleavage and chromosome pairing in Sordaria macrospora

Summary Two recessive mutants spo76 and spo77, altered in U.V. sensitivity, protoplast regeneration and meiotic recombination were isolated in Sordaria macrospora. The suppression of the spo76 phenotype by spo77 suggests that they are involved in the same pathway. The asynaptic spo77 exhibits a rare synaptonemal complex (SC) with abnormally thick and double lateral elements (LE). In spo76, an early centromere cleavage leads to a meiotic arrest after metaphase I; SC are formed, but their discontinuous LE appear to be either unique or split into two thin LE. It is suggested that the corresponding wild-type functions are required for the sister chromatid cohesiveness.     

Synaptonemal complex assembly and H3K4Me3 demethylation determine DIDO3 localization in meiosis

Synapsis of homologous chromosomes is a key meiotic event, mediated by a large proteinaceous structure termed the synaptonemal complex. Here, we describe a role in meiosis for the murine death-inducer obliterator (Dido) gene. The Dido gene codes for three proteins that recognize trimethylated histone H3 lysine 4 through their amino-terminal plant homeodomain domain. DIDO3, the largest of the three isoforms, localizes to the central region of the synaptonemal complex in germ cells. DIDO3 follows the distribution of the central region protein SYCP1 in Sycp3−/− spermatocytes, which lack the axial elements of the synaptonemal complex. This indicates that synapsis is a requirement for DIDO3 incorporation. Interestingly, DIDO3 is missing from the synaptonemal complex in Atm mutant spermatocytes, which form synapses but show persistent trimethylation of histone H3 lysine 4. In order to further address a role of epigenetic modifications in DIDO3 localization, we made a mutant of the Dido gene that produces a truncated DIDO3 protein. This truncated protein, which lacks the histone-binding domain, is incorporated in the synaptonemal complex irrespective of histone trimethylation status. DIDO3 protein truncation in Dido mutant mice causes mild meiotic defects, visible as gaps in the synaptonemal complex, but allows for normal meiotic progression. Our results indicate that histone H3 lysine 4 demethylation modulates DIDO3 localization in meiosis and suggest epigenetic regulation of the synaptonemal complex.     

A change in the phosphorylation pattern of the 30000–33000 M r synaptonemal complex proteins of the rat between early and mid-pachytene

The lateral elements (LEs) of synaptonemal complexes (SCs) of the rat contain major components with relative electrophoretic mobilities (M _r , s) of 30000–33000, which are the products of a single gene. After one-dimensional separation of SC proteins on polyacrylamide-SDS gels, these components show up as two major bands, whereas upon two-dimensional electrophoresis they are resolved in at least 24 spots, which focus at pH 6.5 to 9.5. In this paper we show that these spots represent phosphorylation variants. For the analysis of the phosphorylation of the 30000-to 33000-M _r SC components during progression through meiotic prophase, we developed a procedure for isolation of fractions of testicular cells of the rat that are enriched in separate stages of meiotic prophase. Analysis of the 30000-to 33000-M _r SC components in these fractions by two-dimensional electrophoresis and immunoblotting showed that phosphorylated variants of the 30000-to 33000-M _r SC proteins occur throughout meiotic prophase. However, the extent of phosphorylation changes between early and mid-pachytene, when one phosphate group is probably added to each of the variants.     

Meiosis in gray voles of the subgenus microtus (rodentia, arvicolinae) and in their hybrids

The results of light and electron microscopic (EM) studies of meiosis in Microtus arvalis males of the karyoform "arvalis" (2n = 46, NFa = 80), in hybrids between the chromosomal forms arvalis and obscurus (2n = 46, NFa = 68), in M. rossiaemeridionalis voles (2n = 54, NFa = 54), and in a hybrid between the species M. rossiaemeridionalis and M. kermanensis (2n = 54, NFa = 54) are presented. SC (synaptonemal complex) karyotypes of the parental forms and the hybrids were constructed on the basis of measurements of the length ofautosomal SCs revealed by the EM analysis in spermatocytes at the stage of middle pachytene. The SC karyotypes of M. arvalis and the hybrids female obscurus x male arvalis consist of 22 synaptonemal complexes of autosomal bivalents and the axial elements of the synaptonemal complexes of the sex chromosomes X and Y. The SC karyotypes of M. rossiaemeridionalis and the hybrid M. rossiaemeridionalis x M. kermanensis consist of 26 synaptonemal complexes of autosomal bivalents and a sex bivalent; they differ only in the length of the Y chromosome axis (Y chromosome in the hybrid was inherited from M. kermanensis). Asynaptic configurations of the autosomal SCs were not observed in the hybrids. The SC axial elements of the X and Y chromosomes in the parental forms and in the hybrids were located close to each other throughout pachytene, but they did not form a synaptic region. The normal synapsis in sterile hybrids (M. rossiaemeridionalis x M. kermanensis) and the behavior of the sex chromosomes in meiosis in fertile and sterile hybrids are discussed in the context of specific features of meiosis and reproductive isolation.     

The Centromere Histone Is Conserved and Associated with Tandem Repeats Sharing a Conserved 19-bp Box in the Holocentromere of Meloidogyne Nematodes

Although centromeres have conserved function, centromere-specific histone H3 (CenH3) and centromeric DNA evolve rapidly. The centromere drive model explains this phenomenon as a consequence of the conflict between fast-evolving DNA and CenH3, suggesting asymmetry in female meiosis as a crucial factor. We characterized evolution of the CenH3 protein in three closely related, polyploid mitotic parthenogenetic species of the Meloidogyne incognita group, and in the distantly related meiotic parthenogen Meloidogyne hapla. We identified duplication of the CenH3 gene in a putative sexual ancestral Meloidogyne. We found that one CenH3 (αCenH3) remained conserved in all extant species, including in distant Meloidogyne hapla, whereas the other evolved rapidly and under positive selection into four different CenH3 variants. This pattern of CenH3 evolution in Meloidogyne species suggests the subspecialization of CenH3s in ancestral sexual species. Immunofluorescence performed on mitotic Meloidogyne incognita revealed a dominant role of αCenH3 on its centromere, whereas the other CenH3s have lost their function in mitosis. The observed αCenH3 chromosome distribution disclosed cluster-like centromeric organization. The ChIP-Seq analysis revealed that in M. incognita αCenH3-associated DNA dominantly comprises tandem repeats, composed of divergent monomers which share a completely conserved 19-bp long box. Conserved αCenH3-associated DNA is also confirmed in the related mitotic Meloidogyne incognita group species suggesting preservation of both centromere protein and DNA constituents. We hypothesize that the absence of centromere drive in mitosis might allow for CenH3 and its associated DNA to achieve an equilibrium in which they can persist for long periods of time.     

Gametogenesis and the Chromosomes of Meloidogyne nataliei: Not Typical of Other Root-knot Nematodes

Studies of oogenesis and spermatogenesis revealed that Meloidogyne nataliei is a diploid, amphimictic species with four (n), relatively large chromosomes, and possibly with an XX ♀-XY ♂ mechanism of sex determination. It differs considerably from all other amphimictic, or meiotically parthenogenetic, species of Meloidogyne which have 13-18 smaller chromosomes and from Meloidogyne (Hypsoperine) spartinae which has seven. Consequently, the taxonomic position of M. nataliei needs to be re-evaluated. The chromosomes of M. nataliei and their behavior during gametogenesis resemble more closely chromosomes of the genus Heterodera than those of the genus Meloidogyne. This resemblance, however, may not imply a closer phyletic relationship of M. nataliei to heteroderid nematodes.     

The many facets of SC function during C. elegans meiosis

Sexually reproducing organisms rely on meiosis for the formation of haploid gametes. This is achieved through two consecutive rounds of cell division (meiosis I and II) after one round of DNA replication. During the meiotic divisions, chromosomes face several challenges to ultimately ensure proper chromosome segregation. Unique events unfold during meiosis I to overcome these challenges. Homologous chromosomes pair, synapse, and recombine. A remarkable feature throughout this process is the formation of an evolutionarily conserved tripartite proteinaceous structure known as the synaptonemal complex (SC). It is comprised of two lateral elements, assembled along each axis of a pair of homologous chromosomes, and a central region consisting of transverse filaments bridging the gap between lateral elements. While the presence of the SC during meiosis has been appreciated now for 50 years (Moses, Biophys Biochem Cytol 2:215–218, 1956; Fawcett, J Biophys Biochem Cytol 2:403–406, 1956), its role(s) remain a matter of intense investigation. This review concentrates on studies performed in Caenorhabditis elegans, a powerful system for investigating meiosis. Studies in this organism are contributing to the unraveling of the various processes leading to the formation of the SC and the various facets of the functions it exerts throughout meiosis.The synaptonemal complex-50 years