Telomeres and mechanisms of Robertsonian fusion

The Robertsonian (Rb) fusion, a chromosome rearrangement involving centric fusion of two acro-(telo)centric chromosomes to form a single metacentric, is one of the most frequent events in mammalian karyotype evolution. Since one of the functions of telomeres is to preserve chromosome integrity, a prerequisite for the formation of Rb fusions should be either telomere loss or telomere inactivation. Possible mechanisms underlying the formation of various types of Rb fusion are discussed here. For example, Rb fusion in wild mice involves complete loss of p-arm telomeres by chromosome breakage within minor satellite sequences. By contrast, interstitial telomeric sites are found in the pericentromeric regions of chromosomes originating from a number of vertebrate species, suggesting the occurrence of Rb-like fusion without loss of telomeres, a possibility consistent with some form of telomere inactivation. Finally, a recent study suggests that telomere shortening induced by the deletion of the telomerase RNA gene in the mouse germ-line leads to telomere loss and high frequencies of Rb fusion in mouse somatic cells. Thus, at least three mechanisms in mammalian cells lead to the formation of Rb fusions. Received: 11 November 1997 / Accepted: 21 December 1997     

Chromosomal rearrangements in rock wallabies, Petrogale (Marsupialia: Macropodidae). VII. G-banding analysis of Petrogale brachyotis and P. concinna: species with dramatically altered karyotypes.

G-banded metaphase preparations of cultured fibroblasts were used to construct the karyotypes of Petrogale brachyotis (2n = 18) and P. concinna (2n = 16). The two karyotypes differ significantly from the plesiomorphic karyotype of the genus and from those of all other Petrogale species examined. Petrogale brachyotis and P. concinna are characterised by three synapomorphies: a 1-10 centric fusion, a 3a-6 centric fusion, and a submetacentric chromosome 2 (2s). Both species also possess autapomorphies. Petrogale brachyotis is characterised by submetacentric chromosomes 5 (5s) and 4 (4sm), whereas P. concinna is characterised by a 5-9 centric fusion and a submetacentric chromosome 8 (8m). The 2s, 5s, 4sm, and 8m chromosomes all appear to be derived from their plesiomorphic homologs by centromeric transpositions. Although the rate of chromosome evolution varies considerably in Petrogale, the genus clearly exhibits karyotypic orthoselection, with all the autosomal rearrangements identified being either centric fusions or centromeric transpositions. This study also illustrates the potential for convergent evolution in chromosomally diverse groups and demonstrates the importance of G-banding studies for accurate identification of chromosome rearrangements.     

Centric fusion differences among Oryx dammah, O. gazella, and O. leucoryx (Artiodactyla, Bovidae)

G- and C-banded karyotypes of the genus Oryx were compared using the standard karyotype of Bos taurus. Chromosomal complements were 2n = 56 in O. gazella gazella, 2n = 58 in O. g. beisa and O. g. callotis, 2n = 56-58 in O. dammah, and 2n = 57-58 in O. leucoryx. The number of autosomal arms in all karyotypes was 58. Nearly all variation in diploid number was the result of three independent centric fusions, but one 2n = 57 specimen of O. g. gazella deviated from the normal complement of 2n = 56 due to XXY aneuploidy. A 2;17 centric fusion was fixed in O. g. gazella, whereas O. g. beisa and O. g. callotis lacked this fusion and had indistinguishable karyotypes. Oryx dammah was polymorphic for a 2;15 centric fusion, and O. leucoryx was polymorphic for an 18;19 centric fusion. The five Oryx taxa shared a fixed 1;25 centric fusion; the small acrocentric element involved in the 1;25 fusion was identified by fluorescence in situ hybridization using a cosmid specific to Bos chromosome 25. The X and Y chromosomes were also conserved among the five taxa. Oryx g. gazella differed from the other Oryx species because of the fixed 2;17 centric fusion. This difference reflects an apparently longer period of geographic isolation between O. g. gazella and other populations of Oryx, and it is consistent with the classification of O. gazella and O. beisa as distinct species (see Kingdon, 1997). The lack of monobrachial relationships among the Oryx taxa indicates that sterility barriers between species have not developed. Viability of hybrid offspring constitutes a threat to captive breeding programs designed for endangered species conservation; in the case of Oryx, the 2;15, 2;17, and 18;19 metacentrics could serve as marker chromosomes for assessing hybridization between certain Oryx taxa.     

Mapping of five human chromosome 19 DNA markers to owl monkey chromosomes.

Hybridization of DNA from three panels of karyotypically distinct owl monkey x rodent somatic cell hybrids with human DNA probes resulted in the syntenic assignments of INSR-LDLR-TGFB1-APOE-D19S8 to owl monkey chromosome 25 of karyotype VI (2n = 49/50), INSR-LDLR-TGFB1-D19S8 to chromosome 2 of karyotype II (2n = 54), and INSR-APOE to chromosome 2 of karyotype V (2n = 46). The APOE and D19S8 loci are on adjacent regions proximal to the centromere of chromosomes 25q (K-VI) and 2p (K-II), as determined by in situ chromosomal hybridization analysis. These findings support our previous proposals on (1) the homology of these chromosomes of three owl monkey karyotypes, (2) the evolutionary derivation of chromosome 2 of karyotypes II and V as the result of two separate centric fusion events, and (3) the likelihood that owl monkey chromosome 25 (K-VI) (and its homologs) is a conserved genetic homoeolog of human chromosome 19.     

Robertsonian translocations introduced into an island population of house mice

In April 1982, 77 house mice from the Orkney Island of Eday were released by R. J. Berry and his associates on the Isle of May, Firth of Forth. The Isle of May had a standard house mouse karyotype (2n = 40), while those from Eday are homozygous for three centric fusions (2n = 34). Within 18 months of introduction (September 1983), each centric fusion had increased in frequency from an estimated starting value of 8% to a value close to 50%, and they were segregating in accordance with Hardy–Weinberg expectations. In essence, the transformed population was behaving as a panmictic unit. The frequencies of introduced chromosomes had apparently stabilized by September 1986 with values around 65% for all three fusions. The cytogenetic data obtained in the Isle of May introduction experiment accord well with data for single gene loci (represented by allozyme data) and morphometric data. Male Eday–May F1 hybrids were found to have a low frequency of non-disjunction (13%). This study is unusual because the successful introduction of mice into an established population, and the introgression and stabilization of three centric fusions, could not have been predicted from previous studies on the mouse.     

Chromosomal evolution in Cervidae

On the basis of chromosome data obtained on 30 species and 20 subspecies of Cervidae, a report is submitted on the karyosystematics of this family. The primitive karyotype of Cervidae may be inferred to be composed of 35 acrocentric pairs (2n = 70 FN = 70). During the phyletic evolution of this family different types of chromosome rearrangements were probably selected and the group may have differentiated karyologically into three branches: (1) the Cervinae that fixed a centric fusion resulting in a metacentric pair of autosomes (2n = 68, FN = 70), as shown by the basic karyotype of Cervus elaphus, and where Robertsonian fusions are the preeminent type of chromosome rearrangement; (2) the Odocoileinae, in which pericentric inversions and Robertsonian fusions were favored, yielding first a submetacentric X and then a submetacentric autosome pair. The most representative karyotype is 2n = 70, FN = 74--as in Odocoileus hemionus; and (3) the Muntiacinae, in which centric and tandem fusions were the most common chromosome rearrangements. While Muntiacus reevesi has a karyotype 2n = 46, FN = 46, the chromosome number drops down to 2n = 6 in the females of the M. muntjak vaginalis subspecies group and M. rooseveltorum. Therefore, while the karyotypes are conserved within the subfamilies Cervinae and Odocoileinae; the subfamily Muntiacinae appears to be the most chromosomally diversified group. The few karyological data on the Moschus berezovskii suggest that the Moschinae should be placed in a separate family, the Moschidae.     

Karyotype, centric fusion polymorphism and chromosomal aberrations in captive-born mountain reedbuck (Redunca fulvorufula)

Chromosomes of fourteen captive-born mountain reedbucks (Redunca fulvorufula) have been investigated. The diploid chromosome number was 2n = 56 (FN = 60). The mountain reedbuck karyotype consists of 26 acrocentric and two biarmed chromosome pairs resulting from two centric fusions involving chromosomes 2 and 25, and 6 and 10, respectively. In some animals, 57 chromosomes were detected. Variation in the diploid number was found to be due to polymorphism for the centric fusion 6;10. Both X and Y chromosomes are large and acrocentric. The entire Y chromosome and the proximal part of the X chromosome consist of heterochromatin. The chromosomes X, 9 and 14 appeared to be of caprine type. Chromosome aberrations have been detected in two of the 14 animals investigated. A de novo formed Robertsonian translocation rob(6;13) was found in one female heterozygous for the fusion 6;10. CBG-banding revealed one block of centromeric heterochromatin in the de novo formed translocation rob(6;13) and also in the evolutionarily fixed centric fusions 6;10 and 2;25. One examined male homozygous for fusion 6;10, had a mosaic 56,XY/57,XYY karyotype, with 11% of analyzed cells containing two Y chromosomes. The findings were confirmed by cross-species fluorescence in situ hybridization (FISH) with bovine (Bos taurus L.) chromosome painting probes. The study demonstrates the relevance of cytogenetic screening in captive animals from zoological gardens.     

Karyotype, C-banding, and Ag-NOR analysis in Diplodus bellottii (Sparidae, Perciforms). Intra-individual polymorphism involving heterochromatic regions.

A karyotype analysis was carried out in nine specimens of the Sparid species Diplodus bellottii using conventional staining, as well as C-banding and Ag-NOR banding techniques, showing, respectively, 2n = 46 and fundamental number (FN) = 54, and scarce heterochromatic areas irregularly distributed and up to four NOR active regions that were C positive. When compared with the karyotypes of other related species, one centric fusion giving rise to a large metacentric pair and several pericentric inversions seem to have been involved in the karyotype evolution. An intra-individual polymorphism was detected in one specimen, resulting in two karyotypic forms in roughly identical proportion, owing to a larger C-band by the NOR regions, appearing either in a terminal position of the short arms of pair 2 or in telomeric position of pair 3. These findings suggest that the extra heterochromatic segment responsible for the heteromorphism apparently only involves associated heterochromatin and not the NORs themselves. This C-positive block seems to have eventually been transferred between heterologous NOR chromosomes by a somatic event, facilitated by the physical proximity of NOR pairs in the nucleolus.     

Karyology and phylogeny of some Mesoamerican species of Zamia (Zamiaceae)

Chromosome numbers and karyotypes of four species of Zamia L. (Zamiaceae) are described. Plants of Z. manicata from Colombia are 2n = 18 with eight metacentric (M), four submetacentric (S), two acrocentric (A), and four telocentric (T) chromosomes. Plants of Z. ipetiensis from Panama are 2n = 23 with 3M + 4S + 2A + 14T. Plants of Z. cunaria from Panama have two different chromosome numbers, 2n = 23 with 3M + 4S + 2A + 14T and 2n = 24 with 2M + 4S + 2A + 16T. Plants of Z. acuminata from Costa Rica and Panama are 2n = 24 with 2M + 4S + 2A + 16T. On the basis of the occurrence of a one-to-two-ratio in the variation of M- and T-chromosome numbers in the karyotypes, centric fission or fusion are considered for their potential involvement in the chromosome variation of these plants. Data deriving from morphology and karyology, interpreted in a cladistic framework, suggest that centric fission rather than centric fusion is involved in the karyotype diversification of the four species and their closest Mesoamerican allies.     

Karyotype analysis utilizing differentially stained constitutive heterochromatin of human and murine chromosomes

Constitutive heterochromatin of chromosomes can be visualized utilizing a new differential staining technique which was originally developed by Gall and Pardue (1971). The method facilitates the more certain identification of specific chromosomes within and between cell populations of different origins. Marker chromosomes can be identified in established cell lines over many months of serial passage. Chromosomes of similar morphology within karyotypes of man and mouse can be distinguished in a number of instances. For example, the Y chromosomes of both mouse and man can now be easily detected. The hetero-chromatic staining method also permits discrimination between mouse and human chromosomes in somatic cell hybrids, thus facilitating the assignment of gene markers to chromosomes in somatic cell genetics systems. Instances of translocation of centric heterochromatin to other parts of chromosomes in established tissue culture cell lines are described. An instance of the inheritance of a polymorphic variation in autosomal heterochromatin in man is reported. It is postulated that polymorphisms in the centric heterochromatin may account largely for small heritable chromosome length variations previously described in human populations and termed minor chromosome variants.     

Zaprionus tuberculatus: chromosome map and gene mapping by DNA in situ hybridization.

The genus Drosophila has long been used as a model of karyotype evolution, demonstrating change by paracentric inversion and occasional centric fusion of an ancestral karyotype of five rod-shaped and one "dot" chromosome. This study shows, by mapping D. melanogaster probes hybridized to polytene chromosomes of Zaprionus tuberculatus, that this ancestral pattern extends beyond the genus Drosophila. A formal polytene chromosome map of Z. tuberculatus is presented.     

Chromosome polymorphism in the ant,Pheidole nodus

A survey of chromosome polymorphism was made in populations of Pheidole nodus (Hymenoptera, Formicidae). A total of 1,666 males were collected from 11 localities in Japn. Four polymorphic karyotypes were observed: (1) n = 17 with 4 metacentrics (abbreviated as 4M), (2) n = 18(3M), (3) n = 19(2M) and (4) n = 20(1M). These differences are due to the Robertsonian type rearrangement. The karyotype 18(3M) is found in all the populations examined, but the others are more or less localized in their distribution. The 17(4M) appears mainly in Shikoku and the northern Kyushu populations, 19(2M) along the Pacific coast of Honshu, Shikoku and Kyushu, and 20(1M) in the eastern part of Honshu and Shikoku. This distribution pattern indicates that 18(3M) is the oldest, 19(2M) and 20(1M) are derived from 18(3M) by centric fission, and 17(4M) by centric fusion. The most probable mechanism of karyotype evolution in this species is considered to be the centric fission.     

Karyotype evolution and geographical distribution of the Thai-medaka, Oryzias minutillus, in Thailand

The karyotype of Oryzias minutillus was examined with specimens collected from 18 localities in Thailand. Specimens from the south and the northeast had 2n = 42 acrocentric chromosomes; the arm number (NF) was 42 and NORs-chromosomes were acrocentric type (2n = 42, NF = 42, NORs-A). Specimens from the central and the north were characteristic by having 8-12 large metacentric chromosomes (LM-chromosomes). They had 2n = 28–34 chromosomes, and shared the same NF and NORs-chromosomes of submetacentric type (2n = 34-28, NF = 44, NORs-SM). Specimens from the southeast had 2n = 42 or 40 chromosomes. Their karyotypes had the same NF and NORs-chromosomes as those from the central and the north (2n = 40–42, NF = 44, NORs-SM), though they had no, or only one pair of, LM-chromosomes. The karyotype with 42 acrocentric chromosomes seems to be basic for O. minutillus , and consequently those with NORs-SM and LM-chromosomes seem to be caused through pericentric inversion and centric fusion, respectively. We confirmed that the karyotype evolution had occurred in drainage areas of the Mae Nam Chao Phraya and collaterals (the central, north and southeast). On the other hand, the basic karyotype was preserved allopatrically in the peninsula (the south) and the basin of the Mae Nam Mun, a tributary of the Mekong (the northeast).     

着丝粒横裂和着丝粒并合及其在高等植物染色体进化中的意义

本文讨论了着丝粒横裂和并合及其在高等植物染色体进化中的意义。着丝粒横裂和着丝粒并合是两个矛盾又辩证统一的过程,是染色体的基本变异形式之一,它们同时影响着植物类群的染色体基数、核型对称性、连锁关系、交叉频率和位点等细胞遗传学的重要方面.从而在高等植物染色体进化中起着重要作用,着丝粒和端粒的复制模型为着丝粒的横裂与并合提供了可能的机理,但尚待直接的生物化学证据的证实,原始基数的确定是判别着丝粒横裂与并合的关键。     

Karyotypic study of Echinops (Asteraceae) in Fars Province, Iran

A karyotypic study was performed on 19 Echinops species in Fars Province, Iran. The taxa revealed chromosome counts of 2 n = 28, 30, 32 and 34. Somatic chromosome numbers of 18 species are reported for the first time. Using somatic chromosome number as a criterion for section delimitation, the transfer of E. gedrosiacus, E. cyanocephalm and E. sojakii from Sect. Oligolepis to Sect. Ritropsis is suggested. Differences in basic chromosome numbers point towards the possible role played by centric fusion/fission in the karyotypic evolution of the genus. The Karl Pearson coefficient of correlation and principal components analysis indicated the occurrence of structural changes in chromosomes. The species occupied classes 1A, 2A & 2B of Stebbins' karyotype classification, indicating the presence of a primitive symmetrical karyotype in the genus.     

Cytogenetics studies in Brazilian species of Pseudophyllinae (Orthoptera: Tettigoniidae): 2n(♂)=35 and fn=35 the probable basic and ancestral karyotype of the family Tettigoniidae

The karyotypes of five species of Brazilian Pseudophyllinae belonging to four tribes were here studied. The data available in the literature altogether with those obtained with species in here studied allowed us to infer that 2n(♂)=35 is the highest chromosome number found in the family Tettigoniidae and that it is present in species belonging to Pseudophyllinae, Zaprochilinae and in one species of Tettigoniinae. In spite of that all five species exhibit secondary karyotypes arisen surely by a mechanism of chromosomal rearrangement of centric fusion, tandem fusion and centric inversion types from those with 2n(♂)=35 and FN=35, they share some common traits. The X chromosome is submetacentric (FN=36), heteropicnotic during the first prophase, the largest of the set but its size is rather variable among the species and the sex chromosomal mechanism is of the XO( ♂ ), XX( ♀ ) type. The chromosomal rearrangements involved in the karyotype evolution of the Pseudophyllinae and its relationship with those of the family Tettigoniidae are discussed and we propose that the basic and the ancestral karyotype of the Tettigoniidae is formed by 2n(♂)=35, FN=35 and not by 2n(♂)=31, FN= 31, as usually accepted.     

Isolation of viable reconstituted cells from human karyoplasts fused to mouse cytoplasts.

Long-term survivors of reconstituted human-mouse cells have been isolated and characterized by utilizing nuclear and cytoplasmic genetic markers. Karyoplasts were derived from the human SV40-transformed fetal lung fibroblast strain WI38 VA13, while cytoplasts were obtained from the mouse fibroblast A9 cell line which was both hypoxanthine-aminopterin-thymidine-sensitive (HAT^s; nuclear marker) and chloramphenicol-resistant (CAP^r; cytoplasmic marker). The fusion products were isolated in medium containing HAT and CAP. Clones initially showed a growth pattern different from either human or mouse parental cell, but after repeated subculturing, morphologically resembled the nuclear donor cell. The human and mouse components in these cells were identified from other possible fusion combinations by karyotypic, enzymatic and mitochondrial DNA (mDNA) analyses. The karyotype, using both Q-banding and C-banding revealed only human chromosomes. Electrophoretic mobility of the enzyme malate dehydrogenase, a nuclear controlled enzyme, confirmed the human nucleus. Buoyant density centrifugation of radioactive labelled isolated mitochondrial DNA from the reconstituted cells provided evidence that the cytoplasm was of mouse origin.     

Population cytology of Leptysma argentina Bruner (Leptysminae,Acrididae)

The grasshopper Leptysma argentina possesses a highly polymorphic karyotype which suggests that the species is undergoing a period of intensive chromosomal restructuration and evolution. The two populations studied in this work showed: (1) Polymorphism for a centric fusion between two telocentric autosomes; (2) Polymorphism for an interstitial supernumerary segment in the smallest autosome; (3) Presence of a proximal supernumerary segment in the second smallest autosome; (4) Polymorphism for a B-chromosome; (5) Heterozygosity for a complex spontaneous translocation between autosomes; (6) At least two centric shifts in two pairs of autosomes.Another interesting feature of the cytogenetics of these populations was the occurrence, in a relatively high frequency, of dicentric bridges and acentric fragments in Anaphase I probably due to errors in crossing-over.     

Karyotype Analysis in Lycoris rosea Traub et Moldenke

The present paper embodies the results of a karyotypic analysis for the species Lycoris rosea Traub et Moldenke. The voucher specimen, J. Z. Lin 004 is preserved in the Herbarium of Hanchow Botanical Garden. The chromosome number in root tip cells is found for the first time to be 22, and the karyotype is shown to be an asymmetrical one with rod-shaped chromosomes. A photomicrograph, the karyotype and the idiogram are shown in Figs. 1-2. According to Levan et aL.^[5], the karyotype formula of the species is 2n=22=22t. But based on the classification presented by Bose and Flory^[1], the karyotype formula should be expressed as 2n=22 =C22, and the chromosomes are all with subterminal constrictions. If regarding 11 as the basic number and centric fusion as the major tendency of karyotype evolution as proposed by Inariyama^[2], Stebbins^[6], and Jones^[3,4] in particular, L. rosea would be considered as one of the most primitive species in Lycoris from point of view of karyotype evolution. Reciprocal translocations and centric fusions would give rise to V-shaped chromosomes. Consequently, the successive decrease in chromosome number may have taken place in the speciation of the genus under discussion. Yet further evidence seems ne-cessary for the verification of the speculation.