Mitotic and meiotic chromosomes of Mabuya (Scincidae: Reptilia)

A study was carried out on conventionally stained mitotic and meiotic chromosomes of two species of lizards of the Scincidae: Mabuya caissara and M. macrorhyncha.On the basis of morphological, numerical and metric data, the chromosomes were divided into three groups: two groups of macrochromosomes (A and B) and one of microchromosomes (C). The karyotypes of males and females of both species showed 2n=32 and FN=50 and no differences were detected between the species.     

Mitotic and meiotic chromosomes of the American lobster Homarus americanus (Nephropidae, Decapoda)

The chromosomes of H. americanus have been characterised by C-banding, fluorochrome banding and restriction endonuclease banding. Thanks to these techniques, it has been possible to identify mitotic and meiotic figures clearly and to study the distribution and structure of heterochromatic regions. Moreover, we have identified small supernumerary chromosomes, variable in number and often asynaptic in first meiotic metaphase.     

Mitotic behavior of centromeres in meiosis as the fertility restoration mechanism in wheat-rye amphihaploids

The regulation of chromosomal behavior in meiosis in partly fertile wheat-rye amphihaploids was studied using the centromere specific probes pAWRC1 and Ae. tauschii pAet6-09. Comparative analysis of the probe localization patterns in mitosis, normal meiosis in wheat Triticum aestivum L. and rye Secale cereale L., and meiosis in amphihaploids was performed. The differences in the structure of centromeres in monopolar- and bipolaroriented chromosomes were revealed. Single dense hybridization signals were observed in the diplotene and the metaphase of the first meiotic division, while hybridization signals appeared as stretched bands with diffuse structure located across the centromere region in mitosis and the second round of meiotic division. Based upon the obtained data, we used the corresponding centromere-specific probes as a tool for the analysis of chromosomal behavior in meiosis in amphihaploids. In meiocytes with three types of chromosome behavior (reductional, equational plus reductional, and equational), dense point-like hybridization signals for the pAet6-09 probe were observed for univalents with the reductional division type and stretched bands with diffuse structure for those with the equational division type. Thus, pAet6-09 probe localization patterns suggest some structural and functional specificities of centromeres in the meiosis in wheat-rye amphihaploids that reflect special regulation of chromosomal behavior during equational division. Meiocytes with true mitotic division were also observed in anthers predominantly containing meiocytes with chromosomes undergoing equational division.     

The absence of the somatic association of centromeres of homologous chromosomes in grass mitotic metaphases

Root-tip metaphases from Hordeum vulgare (19 cells), H. marinum (11 cells), Aegilops umbellulata (10 cells) and Zea mays (10 cells) were completely reconstructed from electron micrographs of serially sectioned nuclei. The identity of each chromosome was found by measuring the volumes of its two arms and the presence or absence of a secondary constriction at the nucleolar organising region. With the position of the centromere in three dimensions, these data were used to analyse the relative positions of homologous and heterologous centromeres. In 31 out of the 50 cells analysed, homologues were on average further apart than heterologues. Except for two nucleolar organising chromosomes, there was no evidence of any tendency for the distances between different homologue types to be differently distributed from distances between heterologues. Average distances between homologues of the single nucleolar organising chromosome (linkage group 6) of Zea (2n = 20) were lower than the average for heterologues and the interhomologue distances were distributed significantly differently from the separation distances of chromosome 6 to other chromosomes. Presumably this association occurred because of nucleolar fusion in the previous interphase. Homologues of one of the two nucleolar organising chromosomes of A. umbellulata were also distributed significantly differently from heterologues, with a tendency for homologues to lie farther apart than the average heterologous pair. These results do not support previous work using squashed and spread metaphase preparations (some including abnormal, marked chromosomes) for these species.     

The chromosomes of Micromys minutus (Rodentia, Murinae). II. Pairing pattern of X and Y chromosomes in meiotic prophase

Both light and electron microscopy were used to study the pairing behavior of the sex chromosomes of the harvest mouse, Micromys minutus, in surface-spread pachytene spermatocytes. The XY pairing pattern is very exceptional in that the site of synaptic initiation is located interstitially in the short arms of the X and the Y, next to their centromeric regions. From this tiny euchromatic site, synapsis proceeds unidirectionally along the homologous heterochromatic short arms of the X and the Y toward the ends of the chromosomes. After pairing of the short arm is concluded, synapsis begins between the nonhomologous long arms of the X and the Y in the immediate vicinity of the centromeres and progresses unidirectionally toward the end of the long arm of the Y. A synaptic complex develops between the constitutive heterochromatin of the long arm of the Y and the euchromatin of the long arm of the X. Analysis of C-banded and distamycin A/DAPI-stained diakineses revealed a trefoil-like XY bivalent, which was interpreted to be the result of an interstitial chiasma occurring in the paired short arms of the X and the Y. A conspicuous, electron-dense body, about 1 micron in diameter, was found closely associated with the centromeres of the X and the Y in numerous pachytene spermatocytes. A review of the literature showed that comparable XY-associated bodies have been found in only eight other mammals to date.     

Intraguild predation, cannibalism, and microhabitat use in Calopteryx virgo and Somatochlora metallica larvae: a laboratory experiment

Intraguild predation (IGP) and cannibalism among co-occurring lotic odonate species was studied in Central Finland. A laboratory experiment was performed to assess the microhabitat use and cannibalism between intermediate and late instars of Calopteryx virgo larvae and predation by larger Somatochlora metallica larvae on the intermediate C. virgo instars. The experiment was run in small running-water aquaria where the larvae were able to divide their mutual habitat vertically by clinging onto artificial perches or crawling on the bottom. Life span of the small C. virgo larvae and attack rate on them were compared between the cannibalism and IGP treatments. The effect of predation on the activity, habitat use and spatial distribution of the small C. virgo larvae was examined. The IGP rate was 36%. The prey larvae spent the most of their time on the perches, whereas the S. metallica preferred the substrate. The large C.␣virgo larvae did not cannibalise smaller conspecifics. The presence of a predator (S. metallica) had no effect on the habitat use or activity of the prey (C. virgo) larvae. Habitat use differed more between those species than between conspecifics of different size classes of C. virgo. The spatial distribution between S.␣metallica and C. virgo showed a completely random pattern, whereas the two size classes of C. virgo aggregated in the vegetation. Absence of cannibalism and behavioural observations indicate that C. virgo may have a low tendency for intraspecific aggressions.     

Mitotic and meiotic behavior of the chromosomes of the octet strain of Chlamydomonas reinhardtii

The normal vegetative culture chromosome number of the octet strain of C. reinhardtii was found to be 8. The difficulty of reconciliation of this number with the current genetic map of 16 linkage groups and the conceivability of revision of the map are discussed.Under some cultural conditions unstable vegetative apparent diploids are produced which tend to revert to haploids. It is suggested that in these cases cryptic connections of unknown nature may be retained between endoreduplicated sister elements and that these function for pairing in preparation for reversion to haploidy. Apparently intimate homologous chromosome pairing was found in normal zygotes no older than two or three hours, with light microscope examination. Electron microscope examination revealed paired chromatin elements separated by about 1000 Å at this stage. These may contain rudimentary synaptonemal complexes; none of the usual central region components or lateral elements were evident. Homologous chromosome pairing at this stage is of special interest because the major chromosomal meiotic DNA replication has been reported to occur at a much later stage of development; it has been previously postulated that crossing over precedes this replication, with the four elements necessary for crossing over at the 4-strand stage already present, due to the existence of some form of regular chromatid binemy (or its conceptual equivalent). Observations reported here are consistent with these interpretations.Conceivable relationships of these findings to problems of chromosome behavior in other eukaryotes are discussed.This work was supported by Grant GM 19582 and Research Career Development Award 5-KO3 GM 25988 from the U. S. National Institutes of Health.     

Spindle assembly and cytokinesis in the absence of chromosomes during Drosophila male meiosis

A large body of work indicates that chromosomes play a key role in the assembly of both a centrosomal and centrosome-containing spindles. In animal systems, the absence of chromosomes either prevents spindle formation or allows the assembly of a metaphase-like spindle that fails to evolve into an ana-telophase spindle. Here, we show that Drosophila secondary spermatocytes can assemble morphologically normal spindles in the absence of chromosomes. The Drosophila mutants fusolo and solofuso are severely defective in chromosome segregation and produce secondary spermatocytes that are devoid of chromosomes. The centrosomes of these anucleated cells form robust asters that give rise to bipolar spindles that undergo the same ana-telophase morphological transformations that characterize normal spindles. The cells containing chromosome-free spindles are also able to assemble regular cytokinetic structures and cleave normally. In addition, chromosome-free spindles normally accumulate the Aurora B kinase at their midzones. This suggests that the association of Aurora B with chromosomes is not a prerequisite for its accumulation at the central spindle, or for its function during cytokinesis.     

Mitotic and meiotic analysis in Arctocephalus australis (Otariidae)

The karyotype with C-, G- and NOR-banding of Arctocephalus australis is reported for the first time. The chromosomal number is 2n = 36. The X chromosome, identified in G-banded metaphases from males, is metacentric and the Y chromosome is a minute chromosome, also metacentric. Pachytene spermatocytes were used for synaptonemal complexes analysis with a surface spreading technique. A total of 17 autosomal synaptonemal complexes are observed plus the XY pair. During early pachytene, the X and Y axes are thickened and remain unpaired. As pachytene advances, a short SC is formed between the gonosomes, as it is common among eutherian mammals. The particular asymmetrical appearance of the synaptonemal complex in the sex pair is described and compared to other cases among mammals.     

Mammalian protein SCP1 forms synaptonemal complex-like structures in the absence of meiotic chromosomes

Synaptonemal complexes (SCs) are evolutionary conserved, meiosis-specific structures that play a central role in synapsis of homologous chromosomes, chiasmata distribution, and chromosome segregation. However, it is still for the most part unclear how SCs do assemble during meiotic prophase. Major components of mammalian SCs are the meiosis-specific proteins SCP1, 2, and 3. To investigate the role of SCP1 in SC assembly, we expressed SCP1 in a heterologous system, i.e., in COS-7 cells that normally do not express SC proteins. Notably, under these experimental conditions SCP1 is able to form structures that closely resemble SCs (i.e., polycomplexes). Moreover, we show that mutations that modify the length of the central alpha-helical domain of SCP1 influence the width of polycomplexes. Finally, we demonstrate that deletions of the nonhelical N- or C-termini both affect polycomplex assembly, although in a different manner. We conclude that SCP1 is a primary determinant of SC assembly that plays a key role in synapsis of homologous chromosomes.     

The mitotic, polytene, and meiotic chromosomes of Drosophila ananassae.

The mitotic chromosome complement of D. ananassae consists of four structurally distinguishable submetacentric pairs and all four have been identified with their linkage groups. For the polytene chromosome complement of six arms representing the X, second and third chromosomes, an improved reference map has been constructed and used to describe selected cytogenetically useful rearrangements. In meiotic prophase of spermatocytes, chromosomes 2 and 3 form pachytene-diplotene bivalents whose arms may be associated by chiasmata in postdiplotene stages, but the X, Y and fourth chromosomes participate in a complex multivalent. No correlation was detected between meiotic chromosome behavior and specific genes that regulate crossing over in males. In male inversion heterozygotes having high levels of genetically monitored crossing over, no unequivocal evidence was found for formation of either pachytene inversion loops or anaphase bridges and fragments.     

Aberrations of holokinetic chromosomes and associated lethality after x-irradiation of meiotic stages in Tetranychus urticae Koch (acari, tetranychidae).

Chromosomes of the holokinetic organization type were irradiated with X-rays in various stages of meiosis in unfertilized eggs of Tetranychus urticae Koch. Visible cytological aberrations, lethality and sterility were investigated in subsequent generations. Chromosome fragments are the most frequently occurring light-microscopically visible chromosome aberrations; bridges are not formed. Contrary to expectations, the presence of fragments appears to be positively correlated with the occurrence of lethality; loss of fragments, mis-segregation and the measure of damage to the broken chromosome parts are involved. In contrast with monokinetic chromosomes the earliest lethality occurs only after about 10 divisions. The ratios between different embryonic lethality types (early vs. late) differ depending on the stage irradiated: in more compact chromatin, more serious damage (i.e. more early lethality syndromes) is induced than in less compact chromatin. In the progeny of the surviving males, neither translocations nor independent fragments are found; indirect evidence indicated the occasional presence of inversions. The presumptive inversions are induced more frequently in a chromatin-compact stage (metaphase I) than in a less compact one (telophase I).     

The analysis of exchanges in tritium-labelled meiotic chromosomes

The autoradiographic analysis of exchanges in tritium-labelled meiotic chromosomes is potentially a useful approach to the study of meiotic exchange events since this method differentially labels meiotic chromatids along their entire length. The main problem encountered in earlier autoradiographic studies is that of distinguishing label exchanges generated at chiasmata from label exchanges generated by sister chromatid exchange. This problem was overcome in the present study by the choice of a meiotic system (male meiosis of Stethophyma grossum) where chiasmata are limited to just one proximally localised chiasma in each bivalent. This system allows the positive identification of chiasma-generated label exchanges and demonstrates convincingly the origin of chiasmata through breakage and rejoining of homologous non-sister chromatids. Sister chromatid exchanges are also readily detected in labelled meiotic chromosomes of this species, where they occur with a mean frequency of 0.35 per chromosome. This frequency is similar to that found in mitotic spermatogonial cells and the exchanges are randomly distributed both within and between chromosomes. These features of meiotic sister chromatid exchanges suggest that they are unrelated to non-sister chiasmatic exchanges and they probably have no special meiotic significance.     

Observations of meiotic chromosomes in Gaura (Onagraceae)

Observations of meiotic chromosomes are reported for all 21 species and 3 additional sub species ofGaura (Onagraceae), based upon a study of 647 individuals from 509 naturally occurring populations throughout the range of the genus. The basic chromosome number for the genus isx = 7, and 18 species are diploid withn = 7. Among these, the self-incompatible ones are often highly chromosomally heterozygous, with no homozygous individuals having been found in nature in the perenrennialsGaura lindheimeri andG. villosa, and two-thirds or more of the individuals apparently heterozygous in the following well-sampled species:G. calcicola, G. longiflora, andG. suffulta subsp.suffulta. In contrast, the autogamous species are entirely chromosomally homozygous or nearly so. Two species ofGaura are reported as chromosomal structural heterozygotes, with about 50% pollen abortion:G. biennis andG. triangulata; the translocation systems originated independently of one another. Two of the three polyploid species,G. sinuata andG. drummondii (G. odorata of many authors), are consistently tetraploid (n = 14) and, despite their cytological autotetraploidy, are thought to have originated following interspecific hybridization. They are the only rhizomatous species in the genus and may have had one ancestor in common. The remaining polyploid,G. coccinea, includes populations withn = 7, 14, 21, and 28, as well as evident interploid hybrids and, frequently, supernumerary chromosomes. The relationship among these populations is close and is maintained by frequent hybridization and exchange of genetic material. No other species seems to have participated in their origin, and the association of their chromosomes is consistently that characteristic of autopolyploidy in plants with tetraploid and higher chromosome numbers.     

Misregulation of the kinesin-like protein Subito induces meiotic spindle formation in the absence of chromosomes and centrosomes

Bipolar spindles assemble in the absence of centrosomes in the oocytes of many species. In Drosophila melanogaster oocytes, the chromosomes have been proposed to initiate spindle assembly by nucleating or capturing microtubules, although the mechanism is not understood. An important contributor to this process is Subito, which is a kinesin-6 protein that is required for bundling interpolar microtubules located within the central spindle at metaphase I. We have characterized the domains of Subito that regulate its activity and its specificity for antiparallel microtubules. This analysis has revealed that the C-terminal domain may interact independently with microtubules while the motor domain is required for maintaining the interaction with the antiparallel microtubules. Surprisingly, deletion of the N-terminal domain resulted in a Subito protein capable of promoting the assembly of bipolar spindles that do not include centrosomes or chromosomes. Bipolar acentrosomal spindle formation during meiosis in oocytes may be driven by the bundling of antiparallel microtubules. Furthermore, these experiments have revealed evidence of a nuclear- or chromosome-based signal that acts at a distance to activate Subito. Instead of the chromosomes directly capturing microtubules, signals released upon nuclear envelope breakdown may activate proteins like Subito, which in turn bundles together microtubules.     

The effect of supernumerary chromosomes on meiosis in Puschkinia libanotica (Liliaceae)

The presence of the partly heterochromatic supernumerary chromosomes in pollen mother cells of Puschkinia libanotica raises the chiasma frequency in the A chromosome bivalents. The pattern of chiasma distribution along each of the five A bivalents was related to the DNA labelling pattern of mitotic chromosomes. Regions that showed heavy labelling at the end of the DNA synthetic phase had fewer chiasmata than lightly labelled regions. As this relation is the opposite to that found by Rees and Evans in another species we regard any correlation between labelling pattern and chiasma distribution as fortuitous.     

Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8 by separin

It has been proposed but never proven that cohesion between sister chromatids distal to chiasmata is responsible for holding homologous chromosomes together while spindles attempt to pull them toward opposite poles during metaphase of meiosis I. Meanwhile, the mechanism by which disjunction of homologs is triggered at the onset of anaphase I has remained a complete mystery. In yeast, cohesion between sister chromatid arms during meiosis depends on a meiosis-specific cohesin subunit called Rec8, whose mitotic equivalent, Sccl, is cleaved at the metaphase to anaphase transition by an endopeptidase called separin. We show here that cleavage of Rec8 by separin at one of two different sites is necessary for the resolution of chiasmata and the disjunction of homologous chromosomes during meiosis.     

The structure,meiotic behaviour and effects of B chromosomes in Briza humilis Bieb. (Gramineae)

A single population of Briza humilis contained two types of B chromosome, one a large (B^L) and the other a small (B^S) acrocentric. DNA measurements show that the B^L chromosome contains approximately twice as much DNA per unit length as the members of the regular complement. The meiotic pairing behaviour of the Bs is variable and B^L and B^S are seen to pair in some cells. The presence of B^L depresses the chiasma frequency of the regular complement but the chiasma frequency of A and B chromosomes does not appear to be related. The transmission rate of the B chromosomes is variable and the B^L shows a non-disjunction mechanism during microsporogenesis that is absent during megasporogenesis. For the B^S chromosome the transmission rate is very low and there is no evidence of a non-disjunction mechanism. In general seeds containing B^L chromosomes germinate more slowly than those without B chromosomes.