Holocentric chromosomes of arachnids: Presence of kinetochore plates during meiotic divisions

An ultrastructural study of holocentric chromosomes during meiotic division I and, for the first time, on meiotic division II of three arachnids (the scorpion Tityus bahiensis and the spiders Dysdera crocata and Segestria florentina) is presented. While the results obtained in spiders are similar to those obtained in species previously analyzed, T. bahiensis is an exception to the rule since it shows kinetochore plates during division I. Furthermore, such plates were observed in the three species during division II.     

Ultrastructure,meiotic behavior,and evolution of sex chromosomes of the genus Ellobius

The whole-mount SC preparations from males of three species of the genus Ellobius (Ellobius fuscocapillus, Ellobius lutescens), and Ellobius tancrei were studied by electron microscopy. In the males of Ellobius fuscocapillus, behavioral peculiarities of the sex bivalent (viz. the normal male heterozygosity) are characterized by early complete desynapsis of sex chromosomes (X, Y), occurring at late pachytene-early diplotene. The karyotype of species Ellobius lutescens is unique for mammals. In both sexes it is characterized by an odd number of chromosomes (2n=17). At prophase I the unpaired chromosome 9 is not involved in synapsis with other chromosomes and forms a sex body at the end of pachytene.The complete Robertsonian fan has been described for superspecies Ellobius tancrei. As shown on the basis of G-band patterns the male and female sex chromosomes are cytologically indistinguishable.Analysis of whole-mount SC preparations revealed the formation of a closed sex SC bivalent and showed some morphological differences in the axes of sex chromosomes at meiotic prophase I. A number of assumptions are made about the relationship between the behavior of sex chromosomes, their evolution and the sex determination system in the studied species of genus Ellobius.

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Pairing and segregation of the sex chromosomes in X1X2-males of Dysdercus intermedius with a note on the kinetic organization of Heteropteran chromosomes

The existence of an X₁X₂-mode of sex determination is confirmed by a study of all meiotic stages in the male cotton stainer (X₁X₂ and pertinent stages in the female (X₁X₁ X₂X₂). In the male, the X-chromosomes are heterochromatic and pair end-to-end in early meiotic prophase. At diakinesis, they disjoin and align side-by-side in the center of the spindle, forming a pseudotetrad. Anaphase I is equational for the sex chromosomes. At late anaphase or telophase, X₁ and X₂ join end-to-end but form spindle fiber connections to only one of the poles of the metaphase II spindle, leading to one daughter cell without X chromosomes and one with both X₁ and X₂. An attempt is made to explain sex chromosome pairing and orientation on the basis of a telocentric organization of meiotic chromosomes. The apparent differences in the kinetic organization of mitotic and meiotic chromosomes in Heteroptera are discussed.     

Distance segregation and compound sex chromosomes in mantispids (Neuroptera: Mantispidae)

The sex chromosomes segregate precociously in prometaphase I of male meiosis, without prior synapsis or any physical connection, in 4 species of American mantispids (Neuroptera: Mantispidae). Segregational movements are interpolar, and are implemented through chromosomal fibers. Univalent autosomes, present from diakinesis on in several species, are capable of a similar distance segregation in prometaphase. The sex chromosomes are XX —XY , as is characteristic of the Order, with the exception of Entanoneura phthisica in which both elements are compound —X¹X²X³Y¹Y²Y³in the male, and X¹X¹X²X²X³X³ in the female. In tetraploid sectors of gonial origin in testes of this species no sex bivalents are formed; a distance segregation of 6 sex univalents to each pole is effected, but — as observed in the one individually identifiable pair — segregation separates complete homologues, Y¹ from Y¹, X¹ from X¹, etc. In all species the male meiotic spindle is formed by the collocation of individual chromosomal spindle units within which bivalents become deformed; the timing and degree of deformation vary with the species. In karyotype the American species conform to a common pattern with the known Japanese and European species; diploid numbers range only from 18 to 22, and each complement carries the family insigné of one pair of disproportionately large autosomes in a set of small and rather uniformly sized chromosomes.     

Evolution of karyotype,sex chromosomes,and meiosis in mygalomorph spiders (Araneae: Mygalomorphae)

Spider diversity is partitioned into three primary clades, namely Mesothelae, Mygalomorphae, and Araneomorphae. Mygalomorph cytogenetics is largely unknown. Our study revealed a remarkable karyotype diversity of mygalomorphs. Unlike araneomorphs, they show no general trend towards a decrease of 2n, as the chromosome number was reduced in some lineages and increased in others. A biarmed karyotype is a symplesiomorphy of mygalomorphs and araneomorphs. Male meiosis of some mygalomorphs is achiasmatic, or includes the diffuse stage. The sex chromosome system X₁X₂0, which is supposedly ancestral in spiders, is uncommon in mygalomorphs. Many mygalomorphs exhibit more than two (and up to 13) X chromosomes in males. The evolution of X chromosomes proceeded via the duplication of chromosomes, fissions, X–X, and X‐autosome fusions. Spiders also exhibit a homomorphic sex chromosome pair. In the germline of mygalomorph males these chromosomes are often deactivated; their deactivation and pairing is initiated already at spermatogonia. Remarkably, pairing of sex chromosomes in mygalomorph females is also initiated at gonial cells. Some mygalomorphs have two sex chromosome pairs. The second pair presumably arose in early‐diverging mygalomorphs, probably via genome duplication. The unique behaviour of spider sex chromosomes in the germline may promote meiotic pairing of homologous sex chromosomes and structural differentiation of their duplicates, as well as the establishment of polyploid genomes. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 377–408.     

Chromatin organization in the male germ line of Drosophila hydei

The chromatin organization in developing germ cells of Drosophila hydei males was studied with the highly sensitive DNA stain DAPI (4, 6-diamidino-2-phenylindole dichloride). The prophase of meiosis I is characterized by decondensed chromosomes and only late during this stage do they condense rapidly. The sex chromosomes show allocycly. During postmeiotic development the final condensation of chromatin is preceded by a cycle of condensation and subsequent decondensation. Meiotic chromosomes were studied in more detail after orcein staining. Pairing sites of the sex chromosomes could be localized in the distal end of the heterochromatic arm of the X chromosome and distally in both arms of the Y chromosome. The various heterochromatic parts of the genome condense differentially in meiosis. Chromatin reorganization was studied cytochemically with antibodies raised against histones H1 and H2A of D. melanogaster. The core histone H2A is present in spermatid nuclei until the late elongation stage. However, histone H1 is not found in the chromatin later than the early primary spermatocyte stage. Thus, chromatin reorganization during spermatogenesis in D. hydei is complex. The process is discussed with regard to possible functions.     

A Method for Preparation of Synaptonemal Complexes of Meiotic Chromosomes from Basidial Protoplasts of the White Button Mushroom Agaricus bisporus (Lange) Imbach.

For the first time, preparations of synaptonemal complexes (SCs) were made from meiotic chromosomes of white button mushroom (Agaricus bisporus) basidia. It is the first experience of obtaining SC preparations of filamentous fungi from isolated meiosporangium protoplasts. Previously, only yeast SC preparations were obtained following this approach. The method includes four major stages: isolation of basidium protoplasts by treatment of basidia with lytic enzymes, spreading of protoplast nuclei on a filmy support by osmotic shock, staining the preparations with silver nitrate, and examination under light and electron microscopes. The structures of spread premeiotic nuclei, axial elements of chromosomes, SCs, chromatin, and nucleoli were studied at the leptotene–diplotene stage of meiotic prophase I.     

A comparative study of male meiosis in Drosophila melanogaster and D. virilis

Male meiosis in D. melanogaster cytologically follows the usual pattern, whereas in D. melanogaster and in D. virilis oocytes the chromosomes clump into a karyosphere at early meiotic prophase and remain so up to metaphase I.Male meiosis in D. virilis spermatocytes has an intermediate character: a part of the chromatin clumps together in a karyosphere at early prophase, whereas the other part of the chromatin remains diffuse all through prophase. At the end of prophase, the diffuse chromatin becomes integrated into the karyosphere before metaphase I. During the meiotic divisions the chromosomes have the same clumped aspect as those in Drosophila oocytes and thus differ strikingly from the dividing chromosomes in D. melanogaster spermatocytes.In D. virilis spermatocytes the nucleolus exhibits changes during the meiotic prophase that may be related to synthetical activities. The DNA specific staining with the fluorochrome DAPI reveals the existence of extrachromosomal DNA in the later prophase. Other striking differences in meiotic events between the two Drosophila species concern the centrioles and spermiogenesis.     

The meiotic structure and behavior of the strongly heteromorphic X/Y sex chromosomes of neotropical plethodontid salamanders of the genus Oedipina

Plethodontid salamanders in the genus Oedipina are characterized by a strongly heteromorphic sex-determining pair of X/Y chromosomes. The telocentric X chromosome and the subtelocentric Y chromosome are clearly distinguished from the autosomes and their behavior during meiosis can be sequentially followed in squash preparations of spermatocytes. In Oedipina the sex chromosomes are not obscured by an opaque sex vesicle during early meiotic stages, making it possible to observe details of sex bivalent structure and behavior not directly visible in other vertebrate groups. The sex chromosomes can first be distinguished from autosomal bivalents at the conclusion of zygotene, with X and Y synapsed only along a short segment at their non-centromeric ends, forming a bivalent that contrasts sharply with the completely synapsed autosomes. During pachytene, the XY bivalent becomes progressively shortened and more compact, disappearing as a visible structure when pachytene progresses into the diffuse stage of male meiosis. Diplotene bivalents gradually emerge from the diffuse nuclei, presumably by the return of the loops of chromatin into their respective chromomeres. During early diplotene, the X/Y bivalent is clearly visible with a single chiasma within the synapsed segment. This chiasma is terminalized by first meiotic metaphase with the X and Y appearing either in end-to-end synaptic contact or as univalents separated at opposite poles relative to the equatorially distributed autosomal bivalents. In C-banded preparations, the Y is entirely heterochromatic while the X contains a large centromeric C-band and another block of heterochromatin located at the telomeric end, in the region of synapsis with the Y. We find no cytological evidence of dosage compensation, such as differential staining of the X chromosomes or Barr bodies, in mitotic or interphase cells from female animals.     

Telomere and centromere DNA are associated with the cores of meiotic prophase chromosomes

Mouse (Mus musculus) whole-mount, surface-spread, meiotic prophase chromosomes have an axial which extend chromatin loops. This arrangement permits a novel approach to the analysis of chromosome structure. Using in situ hybridization, the types of DNA sequences preferentially associated with the SC and the types located primarily in the chromatin loops can be determined. With biotinylated probes, detected by avidin conjugated to FITC, we present evidence for differential chromatin-SC interaction. The telomere sequence (TTAGGG)_n is associated exclusively with the two ends of each autosomal SC rather than with the chromatin loops. The minor satellite DNA sequences are predominantly localized to the centromeric region of the SC, as defined by CREST serum anti-centromere antibodies. In contrast, the major satellite DNA probe hybridizes to the chromatin loops of the centromeric heterochromatin, and a probe containing a LINE sequence hybridizes to chromatin loops in general with no obvious preference for the SC. These observations demonstrate that, depending on the type of DNA sequence, the chromatin has different properties in regard to its association with the SC.D.P. Bazett-Jones     

A method for preparation of synaptonemal complexes of meiotic chromosomes from basidial protoplasts of the common mushroom Agaricus bisporus (Lange) Imbach

For the first time, preparations of synaptonemal complexes (SCs) were made from meiotic chromosomes of white button mushroom (Agaricus bisporus) basidia. It is the first experience of obtaining SC preparations of filamentous fungi from isolated meiosporangium protoplasts. Previously, only yeast SC preparations were obtained following this approach. The method includes four major stages: isolation of basidium protoplasts by treatment of basidia with lytic enzymes, spreading of protoplast nuclei on a filmy support by osmotic shock, staining the preparations with silver nitrate, and examination under light and electron microscopes. The structures of spread premeiotic nuclei, axial elements of chromosomes, SCs, chromatin, and nucleoli were studied at the leptotene-diplotene stage of meiotic prophase I.     

Meiotic differences between three triatomine species (Hemiptera,Reduviidae)

We have found the following differences in the male meiosis among three triatomine species: (1) The three largest autosomal bivalents ofTriatoma infestans are heterochromatic.Rhodnius prolixus has two autosomal bivalents with heterochromatic blocks.Triatoma rubrovaria does not show any heteropycnotic autosomes. (2) Sex chromosomes inT. infestans form a chromocenter. At early prophase terminal associations are seen between sex chromosomes inT. rubrovaria, and they maintain a close association until diakinesis. An intimate association between the X and Y chromosomes is observed during early prophase inR. prolixus, but a distant association is maintained by the sex chromosomes at diffuse and diplotene stages in this species. (3) Polyploid nuclei of the nutritive cells are quite distinct. Numerous chromocenters of different shapes and sized are seen in those ofT. infestans. InT. rubrovaria one chromocenter having two positively heteropycnotic elements is observed surrounded by homogeneous chromatin. Only one compact chromocenter is found amongst unevenly distributed chromatin, inR. prolixus.     

Comparative studies on sex chromosomes in different species

InLocusta migratoria (XO),Mus musculus, Rattus norvegicus, Mesocricetus auratus, Cricetulus griseus andHomo sapiens typical sex vesicle structures are visible in early meiotic prophase stages up to pachynema. The structures include whole sex chromosomes or parts thereof. The heterologous parts and the solitary X chromosome ofLocusta pass diplonema, diakinesis and first metaphase nearly in mitotic shape. Entirely heterologous sex chromosomes are kept together by a unilateral and achiasmatic end connection. The sex vesicle is interpreted as a special structure of allocyclic sex chromosomes or parts of them, corresponding in early meiotic stages to the chromocenters of mitotic interphase nuclei. The formation of the sex vesicle is independent of the orthoploidy of nuclei and of the DNA ratio between autosomes and sex chromosomes. Heteropycnotic behaviour of sex chromosomes in spermatids is interpreted as a condition capable of blocking genetic activity, like in the Barr bodies of female somatic nuclei, giving equal chances of fertilization to both types of gametes.Based on a paper read at the XI International Congress of Genetics, of which an abstract has appeared in the congress proceedings, Genetics Today, Vol. 1, p. 299 (1963).     

Reorganization and condensation of chromatin in mitotic prophase nuclei of Allium cepa

This paper studies the process and features of chromosome construction in mitotic prophase cells of Allium cepa. The results showed that a prominent reorganization of chromatin occurred during G₂-early prophase. The 250–400 nm thick compact chromatin threads in G₂ nuclei began to disorganize into about 30, 100 and 220 nm chromatin fibres which constituted the loosely organized chromosome outlines in early prophase before chromosome condensation. In middle prophase, chromosome condensation was characterized by the formation of many condensed regions (aggregates of chromatin), which increased in size (1–1.5 m) when prophase proceeded. Meanwhile, the chromatin threads that constituted and connected the condensed regions became increasingly thicker (120–250 nm). In late prophase adjacent condensed regions fused to form cylinder-shaped chromosomes. Based on these observations, we come to the conclusion that the construction of prophase chromosomes is a two-step process, that is, the reorganization and condensation of chromatin. In addition, we report the study of silver-stained, DNA- and histone-depleted prophase chromosomes, describe morphological features of the non-histone protein (NHP) residue in early, middle and late prophase chromosomes, and discuss the roles of NHPs in chromosome construction.     

Behavior of sex chromosomes at early meiosis stages in three wood mice species of the genus <Emphasis Type="Italic">Apodemus</Emphasis> (Rodentia,Muridae)

The prophase of the first meiotic division was studied in field mice of the species Apodemus (Sylvaemus) flavicollis, A. (S.) ponticus, and A. (S.) uralensis by light and electron microscopy. The karyotypes of three species were described on the base of electron microscopy of synaptonemal complexes in spermatocytes I. The axial elements of the sex chromosomes at early-middle pachytene synapse along the major portion of the Y axis; at late pachytene-early diplotene, the synapsis region shrinks; and at diakinesis-metaphase I, X and Y chromosomes associate end-to-end in all species studied. The behavior of sex chromosomes in the synapsis in the species studied was quite uniform. The results are discussed in the context of earlier data on the behavior of sex chromosomes in various rodent species in meiosis prophase I and their banding.     

The meiotic behaviour of the XY pair in Lutreolina crassicaudata (Marsupialia: Didelphoidea)

The meiotic behaviour of the XY pair of the didelphid Lutreolina crassicaudata is analyzed by microspreading of spermatocytes for visualization of chromosomal axes and by three-dimensional reconstruction of spermatocyte nuclei from EM thin sections. The delay in pairing of sex chromosomes compared to autosomes and the absence of a synaptonemal complex between the axes of the X and Y chromosomes, already described for South American marsupials by three-dimensional reconstruction and for Australian species with synaptonemal complex microspreadings, is confirmed for this species. Sections demonstrate that at the diffuse stage and diplotene the dense plate occupies the region of the inner face of the nuclear envelope in contact with the XY body. Spreads show an structure similar in staining to the axes that becomes apparent simultaneously with the dense plate, called a balloon. The mechanism of XY pairing during meiotic prophase appears to be common to American and Australian marsupials as the same morphological pattern is found in all the species described. This mechanism is different from the way of pairing and segregation known for eutherian sex chromosomes.     

Synapsis,desynapsis, and formation of polycomplex-like aggregates in male meiosis of Pales ferruginea (diptera,tipulidae)

Meiotic prophase in Pales ferruginea spermatocytes was studied by means of 3D electron microscopical reconstruction. Chromosomes in early prophase nuclei from freshly hatched IVth instar larvae were found to be partially synapsed at several sites along the genome. The synaptic regions are distributed more or less homogeneously throughout the nucleus, i.e., they are not preferentially located. The average lengths of the synaptic regions (length of synaptonemal complex fragments, SC) were 0.62, 0.73, 0.86, and 1.0 m in four different nuclei. Unpaired axial cores were not observed, neither in nuclei with partially synapsed chromosomes nor in nuclei devoid of SC fragments. — Chromosomes in diplotene nuclei from 7–8 days old IVth instar larvae were also found to be partially synapsed, revealing SC fragments with average lengths of 1.6 and 1.95 m in two nuclei analysed. The longest SC fragments observed in diplotene were 3–6.5 m. Diplotene SCs show signs of disintegration. Unpaired axial cores do not occur. The number and the average length of SC fragments decreases towards early diakinesis. During this stage the formation of polycomplex-like aggregates (PC) begins. In later diakinesis each nucleus contains one (occasionally two) PC, while SC fragments are absent. — The observations were interpreted as follows: 1. Due to the absence of unpaired cores early prophase in Pales is difficult to relate to the typical stages of lepto- and zygotene as observed in other organisms. Synapsis seems to begin at many sites along the chromosomes. Since zipper-like alignment of cores does not occur, the entire SC structure evidently becomes assembled de novo during synapsis. 2. During desynapsis in diplotene the SCs seem to become gradually distintegrated into molecular subunits up to diakinesis. 3. The integration of SC material into PCs in diakinesis may be understood as a crystallization process from a pool of molecular subunits.     

Metaphase and meiotic chromosomes,synaptonemal complexes (SC) of the lizard <Emphasis Type="Italic">Zootoca vivipara</Emphasis>

Somatic mitotic and meiotic chromosomes at the pachytene and at the metaphase I of the males of the viviparous lizard, Zootoca vivipara (Lichtenstein, 1823), from northwestern Russia, belonging to the Russian form of Z. v. vivipara, are examined. The spreading of synaptonemal complexes (SC) of their chromosomes are obtained and analyzed for the first time. Eighteen SC are observed, including SC of the Z₁Z₁ (pairs 5 or 6) and the Z₂Z₂ (pair 13) sex chromosomes. Characteristics of SC are compared with the number and the shape of bivalents and with those of the karyotype structure. In the studied Russian form of Z. v. vivipara, the length ratios of bivalents correlate with that of mitotic chromosomes (2n = 36); however, some specificity in the morphology of SC of the Z₁Z₁ sex chromosomes is reported in this article.     

Identification and patterns of synapsis of the autosomally translocated Y-chromosome of the Indian mongoose,Herpestes auropunctatus (Hodgson)

The multiple sex chromosome system, X₁X₂Y /X₁X₁X₂X₂, in the small Indian mongoose, Herpestes auropunctatus, results from a translocation of a part of Y chromosome to an autosome. It is not possible to distinguish the autosome which harbours the Y chromosome element in the somatic complement. By employing the surface-spreading technique to prophase I meiocytes we have identified the region to which the Y chromosome has been translocated as the short arm of chromosome 9 which is a subtelocentric chromosome. This Y chromosome component lacks heterochromatin and no sex vesicle is organised during meiotic prophase. This suggests to us that Y heterochromatin in mammals may be required for the production of a sex vesicle.We take great pleasure in dedicating this paper to our revered teacher Prof. S.P. Ray-Chaudhuri, who initiated us to the field of Cytogenetics, on the occasion of his 75th birth day