Meiotic chromosome structure. Kinetochores and chromatid cores in standard and B chromosomes of Arcyptera fusca (Orthoptera) revealed by silver staining.

The behaviour of two chromosome structures in silver-stained chromosomes was analyzed through the first meiotic division in spermatocytes of the acridoid species Arcyptera fusca. Results showed that at diakinesis kinetochores and chromatid cores are individualized while they associate in bivalents of metaphase I; only kinetochores and distal core spots associate in the sex chromosome. Metaphase I is characterized by morphological and localization changes of both kinetochores and cores which define the onset of anaphase I. These changes analyzed in both autosomes and in the sex chromosome allow us to distinguish among three different substages in metaphase I spermatocytes. B chromosomes may be present as univalents, bivalents, or trivalents. Metaphase I B univalents are characterized by separated cores except at their distal ends and individualized and flat sister kinetochores. At anaphase I sister kinetochores of lagging B chromatids remain connected through a silver-stained strand. The behaviour of cores and kinetochores of B bivalents is identical with that found in the autosomal bivalents. The differences in the morphology of kinetochores of every chromosome shown by B trivalents at metaphase I may be related to the balanced forces acting on the multivalent. The results show dramatic changes in chromosome organization of bivalents during metaphase I. These changes suggest that chromatid cores are not involved in the maintenance of bivalents. Moreover, the changes in morphology of kinetochores are independent of the stage of meiosis but correlate with the kind of division (amphitelic-syntelic) that chromosomes undergo.     

Dosage effects on morphological and quantitative traits in maize aneuploids.

Dosage effects generated by either loss or gain of a chromosome segment were used to identify chromosome regions associated with morphological and quantitative characters in maize (Zea mays L.). Using B-A translocation stocks introgressed into a B73Ht background, a chromosome arm dosage series in a Mo17Ht x B73Ht F1 hybrid background was created for 18 of the 20 chromosome arms. The dosage series was then evaluated for 12 quantitatively inherited characters to associate specific phenotypic changes in a trait with a specific chromosome arm. Not only did our results show the familiar aneuploid syndrome phenomenon, but differential dosage effects among particular chromosome arms were demonstrated. All the quantitative traits measured and all the chromosome arms examined in this study were responsive to changes in chromosome arm dosage. The possible bases behind those differences and their utility in identifying quantitative trait loci, as well as the genetic relationships among the group of quantitatively inherited characters studied, are considered. Key words : corn, chromosome arm, B-A translocations, dosage analysis.     

Significance of mitotic spindle checkpoint genes in leukemia

Leukemia is a clonal proliferative disorder of a multipotent hematopoietic stem cell that leads to abnormal cell growth and/or differentiation. The hallmark of the disease is the presence of oncogene expression in bone marrow or peripheral blood resulting from chromosome translocations. The development of leukemia and the progression of the disease are multistage processes implicated in a series of molecular changes preceeded chromosomal instability and aneuploidy. The most likely of the above-mentioned changes include the following: the appearance of additional chromosome translocations, the activation of other oncogenes not expressed previously, the loss of tumor suppressor genes, abnormal centrosome duplication, and the dysfunction of genes that coordinate accurate chromosome alignment and chromosome segregation during mitosis. The last two molecular events controlled by mitotic spindle checkpoint genes play a role in leukemogenesis and are probably involved in apoptosis.     

Advances in the detection of ploidy differences in cancer by in situ hybridization.

Three main techniques allow the detection of changes in the cellular genomic content. The karyotyping procedure on metaphase spreads can give specific information on chromosome number and structural chromosome changes, but analyses are restricted to a limited number of chromosome spreads. Furthermore, cell culturing of (in particular solid) cancer specimens can result in selection of a minor tumour cell population with a high proliferative capacity. On the other hand, flow cytometry allows the analyses of large numbers of cells, but does not detect small variations in the DNA content or structural changes. The fluorescent in situ hybridization (FISH) procedure combines the advantages of the two former procedures, in that relatively large numbers of cells can be analysed easily and specific chromosomal changes can be detected.     

Radiation of chromosome shuffles

Rock wallabies, Petrogale, exhibit chromosome diversity that is exceptional in marsupials, with 20 distinct chromosome races being recognized. Many of the karyotypic changes identified within Petrogale appear to be recent, although the rate of chromosome evolution varies between taxa. While the patchy distribution of Petrogale and their social structure would facilitate the fixation of novel rearrangements, these factors alone do not explain the pattern of chromosome evolution shown in this group. The chromosome changes that have come to characterize each taxon may offer selective advantages in the particular areas occupied, or it may be that these rearrangements play an important role in reproductive isolation. In Petrogale, the taxa with the largest number of chromosome rearrangements are those that are sympatric, or have multiple zones of parapatry, with other members of the genus. Male hybrids from a variety of chromosomal admixtures were found to be sterile, but with those heterozygous for the least complex rearrangements being least affected. As expected, equivalent female hybrids were less severely affected. Chromosomal and genic changes both appear important in these processes.     

Chromosomal Evolution in the Genus Brachyscome (Asteraceae, Astereae): Statistical Tests Regarding Correlation Between Changes in Karyotype and Habit Using Phylogenetic Information

Brachyscome and 8 taxa of its allied genera, Australian Astereae. Statistical tests regarding correlations between changes in chromosome number, total chromosome length, mean chromosome length, karyotypic asymmetry and chromosome length heterogeneity and changes in habit were performed based on the matK molecular phylogenetic tree. The reductions in chromosome number and total chromosome length, and the increases in mean chromosome length, chromosome length heterogeneity and karyotypic asymmetry were found to be correlated with the change in habit from perennial to annual. A reduction in total chromosome length is favored to shorten the mitotic cell cycle and to produce smaller cells conducive to more rapid development of smaller annuals under the time-limited environment. Stepwise dysploidal reductions in chromosome number were achieved through the translocation of large chromosome segments onto other chromosomes, followed by the loss of a centromere, resulting in one fewer linkage group and one fewer haploid chromosome. The correlations between the dysploidal reduction in chromosome number and the increases in mean chromosome length, length heterogeneity and asymmetry in karyotype can be attributed to this mode of chromosomal change. These changes occurred independently in several different lineages in Brachyscome. Received 27 May 1998/ Accepted in revised form 18 January 1999     

Karyological studies on some species of the families Echinostomatidae and Plagiorchiidae and aspects of chromosome evolution in trematodes

The morphometric characteristics of the chromosomes and the variability of the C-heterochromatin blocks in the trematodes Echinoparyphium aconiatum, E. recurvatum, Echinostoma revolutum, E. echinatum, Hypoderaeum conoideum, Isthmiophora melis, Paryphostomum radiatum, Neoacanthoparyphium echinatoides, Plagiorchis maculosus and Opisthioglyphe ranae are determined. The terminal and subterminal localisation of the centromere is a characteristic of the taxa examined. Typical two-arm chromosomes are rare. The karyotype of the examined trematodes is asymmetrical, and this asymmetry is a result of differences in the lengths of the chromosome arms. It is proved that the regression of the lengths of the chromosome arms has a linear character with a similar angle of slope in the different species. Centric fusion and unreciprocal translocations are accepted as contributing significantly to chromosome changes. A hypothesis on the possible mechanism of chromosome changes in the trematode karyotype is proposed.     

CyDAS: a cytogenetic data analysis system

For statistical analyses in cancer cytogenetics, the genomic changes encoded by the karyotype must be translated into numerical codes. We developed a program, which extracts chromosomal gains and losses as well as breakpoints from the karyotype. The changes are compiled in tables according to the chromosome bands involved and/or depicted in projection to the respective chromosome ideogram. The data are ready to be integrated into further statistical analyses. The program may be run as desktop or Internet application.     

The occurrence of the Philadelphia chromosome in essential thrombocytosis

Summary The Philadelphia chromosome, assessed with banding techniques, was detected in 98.3% of bone marrow cells of a 46-year-old black female presenting with essential thrombocytosis. The patient has been followed for the past two years with no signs of chronic myelocytic leukemia. Her platelet counts remain elevated, and she shows no other hematologic changes. Comparisons with the 21q- marker associated with thrombocytosis are made. The role of the Ph' chromosome in myeloid malignant changes and the implications of the present findings in thrombocytosis are discussed.     

Karyotype evolution in the genus Boronia (Rutaceae)

New chromosome counts are reported for Boronia clavata 2 n  = 14, B. heterophylla 'Near White' 2 n  = 15, B.  'Carousel' 2 n  = 16, B. deanei 2 n  = 22, B. chartacea 2 n  = 32, B. keysii 2 n  = 32, B. pilosa 2 n  = 44, B. anethifolia 2 n  = 36 and B. citriodora 2 n  = 108. Studies in 20 genotypes of 18 species and one interspecific hybrid revealed that they are highly complex in terms of chromosome number, ploidy level, chromosomal length, karyotype constitution and asymmetry. Karyotype analysis indicated that Boronia taxa with high chromosome numbers are primitive and those with lower numbers are derived. The basic chromosome number for this genus is suggested to be x = 18. Analysis of chromosome number, variations of total chromosome length (TCL) and average chromosome length (ACL), Nombre Fondamental (NF) and karyotype asymmetry suggest that dysploid reduction is the major mechanism in Boronia karyotype evolution. Chromosomal rearrangements might also have been involved. Origin, chromosome number changes and spread of Boronia are discussed in relation to the species divergence and the geological and climatic changes of the Australian continent.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 309–320.     

Cytogenetic markers in hematoproliferative disorders

In the last decade the improvement of methods of chromosome analysis has allowed new insights into the correlation of specific chromosome changes and certain types of malignant hematologic disorders. Even if a clear-cut correlation between a certain chromosomal marker and a certain malignancy is the exception, it is well established that specific chromosome aberrations occur nonrandomly in specific tumors. Moreover, it has been shown that so-called cellular oncogenes are located on those chromosome regions which are involved in translocations and other structural chromosome abnormalities in particular malignant tumors. The significance of chromosome alterations in leukemias and lymphomas is illustrated by examples concerning well-established data, on the one hand, and findings which have still to be confirmed, on the other. This may demonstrate that human tumor cytogenetics are a dynamically and vigorously developing branch of cancer research.     

Clonal structural chromosomal rearrangements in lymphocytes of four patients with Werner's syndrome

Multiple numerical and structural chromosome abnormalities were found in cultured lymphocytes of four patients with Werner's syndrome. The proportion of metaphases with structural and/or numerical aberrations varied from 30 to 44% and several of them were clonal. These results confirm definitively that Werner's syndrome is a chromosome rearrangement syndrome and that these non-constitutional chromosome changes are not exclusive of cultured fibroblasts but present also in lymphocytes.     

Molecular organisation of the plant genome: Its relation to structure,recombination and evolution of chromosomes

Large scale changes in nuclear DNA amount accompany the evolution of species of higher plants. Much of the nuclear DNA accrued during the evolution of species does not encode genetic information and is selectively neutral. Nonetheless, the pattern of distribution of the excess DNA within and between chromosome complements suggests that there are rigid constraints underlying evolutionary changes in genome organisation. A five-fold increase in the amount of nuclear DNA has occurred in the evolution ofLathyrus species. Not withstanding this massive DNA variation, species show consistently similar patterns in base sequence proliferation, divergence and DNA distribution within and between chromosome complements. Within chromosome complements, the excess DNA is distributed evenly in all chromosomes irrespective of the large differences in chromosome size and, between complements, DNA distribution is discontinuous; species cluster into DNA groups with remarkably regular intervals. Similar constraints govern the frequency and distribution of chiasmata in the chromosome complements. Between species chiasma frequency and nuclear DNA amounts are not correlated but within complements it is positively correlated with the amount of DNA contained in each chromosome.     

Different subfamilies of alphoid repetitive DNA are present on the human and chimpanzee homologous chromosomes 21 and 22.

The alphoid repeat DNA on chimpanzee chromosome 22 was compared with alphoid repeat DNA on its human homologue, chromosome 21. Hybridization of different alphoid probes under various conditions of stringency show that the alphoid repeats of chimpanzee chromosome 22 are not closely related to those of human chromosome 21. Sequence analysis of cloned dimer and tetramer EcoRI fragments from chimpanzee chromosome 22 confirm the low overall level of homology, but reveal the presence of several nucleotide changes which are exclusive to the chromosome 21 subfamily of human alphoid DNA. Southern blot analysis of alphoid repeat DNA on the chimpanzee X chromosome suggests this subfamily has been strongly conserved during and since the separation of chimpanzee and man although the two subfamilies can be distinguished on the basis of Taq I restriction fragments.     

Protocols for cytogenetic studies of human embryonic stem cells

All cultured cells develop chromosome changes over time, including cultures of human embryonic stem cells (hESC), but only those cells with adaptive chromosomes changes survive. The most frequent chromosome changes in hESC cultures are trisomy 12 and trisomy 17. Cells with these trisomies are indistinguishable from normal cells by appearance and also demonstrate typical markers of pluripotency, making them difficult to identify without cytogenetic analysis. Early detection of these cells is essential since cells with trisomy 12 and 17 can replace the normal cell population in 5-10 passages. Cytogenetic analysis using G-banding is considered to be the gold standard for detecting chromosome abnormalities and, when used in combination with interphase FISH, provides a sensitive method for early detection of cytogenetic aberrations, such as full and partial trisomies of chromosomes 12 and 17. The following discussion describes the cytogenetic methods used in our laboratory to study cultured hESCs, along with recommendations for integrating these methods into a plan for routine cell line quality control.     

Chromosomal rearrangements,genome reorganization,and speciation

Historical analysis of studying chromosome changes in evolution allows better understanding of the current level of research in this area. Reorganizations of the genetic system due to chromosomal rearrangements have important evolutionary consequences and may lead to speciation. Despite the complexity of evaluating the primacy of chromosome changes in speciation events, such phenomena are possible and occur in nature, as recent studies have demonstrated.     

Significance of chromosome 5 and 17 changes in the development of carcinoma of the cervix uteri

Two chromosomes that undergo nonrandom changes in carcinoma of the cervix and have been studied for several decades in this laboratory are discussed. The first, chromosome 5, is discussed in view of the frequent appearance of an isochromosome for 5p, often in two or more copies and commonly associated with fewer that the expected number of normal copies of this chromosome. The second is chromosome 17, where a translocation involving another chromosome may result in a 17p+, and the significant change appears to be a loss from 17p that may include the p53 gene (TP53) and/or other tumor-suppressor genes located on this chromosome arm.     

The long and the short of avian W chromosomes: no evidence for gradual W shortening

The well-established view of the evolution of sex chromosome dimorphism is of a gradual genetic and morphological degeneration of the hemizygous chromosome. Yet, no large-scale comparative analysis exists to support this view. Here, we analysed karyotypes of 200 bird species to test whether the supposed directional changes occur in bird sex chromosomes. We found no support for the view that W chromosomes gradually become smaller over evolutionary time. On the contrary, the length of the W chromosome can fluctuate over short time scales, probably involving both shortening and elongation of non-coding regions. Recent discoveries of near-identical palindromes and neo-sex chromosomes in birds may also contribute to the observed variation. Further studies are now needed to investigate how chromosome morphology relates to its gene content, and whether the changes in size were driven by selection.