A cytogenetic study on congenital limb malformations in the Japanese monkey (Macaca fuscata)

A cytogenetic investigation was performed on 88 Japanese monkeys (Macaca fuscata) with abnormal limbs from 11 free-ranging provisioned troops including nine individuals with abnormalities indistinguishable as to whether they were congenital or injurious. All of the monkeys with abnormal limbs including the nine questionable individuals had the same karyotypes as those of normal individuals. The chromosome number was 42, consisting of 20 bi-arm autosome pairs and a submetacentric X-chromosome and Y-chromosome. The ninth chromosome pair, which was the only chromosome pair with remarkable secondary constriction, displayed length polymorphism of the centromeric C-band and secondary constriction in both deformed and normal monkeys. These kinds of variants have also been commonly found in other monkey species, which have almost the same karyotype as the Japanese monkey and have not been reported to show frequent occurrence of limb malformation. We concluded therefore that chromosomal abnormalities could be excluded from the main causal factors for limb malformations of the Japanese monkey.     

Chromosome mapping of Xenopus tropicalis using the G- and Ag-bands: tandem duplication and polyploidization of larvae heads

Developmental cytogenetic analyses of Xenopus tropicalis larvae from two origins were performed on stage 27-34 heads treated with colchicine. Standard G-band karyotyping using trypsin and chromosome mapping of 184 bands were examined. Although the main karyotype was 2n = 20, polyploidy (3n = 30 or 4n = 40) and aneuploidy were detected in each individual treated with colchicine, even those treated for only 1 h. The percentage of polyploid karyotypes was 10-20% across the total of measured metaphases. The mean mitotic index was 0.10. Chromosomal breaks and exchanges were detected at the secondary constriction of chromosomes 5 or 6. Ag-band detection showed clearly positive staining at the secondary constriction of chromosome 5, which corresponds to the nucleolar organizer region. Tandem duplication of negative G-bands at the secondary constriction of chromosome 6 and the short arm of chromosome 10 was suggested by this study. X. tropicalis thus provides a good model to study the mechanism and effects of chromosomal abnormalities, gene mapping and tissue specific gene expression in the developmental process.     

Differentiation of chromosome banding sequence pools and genomic dna in Holarctic natural populations of <Emphasis Type="Italic">Chironomus entis</Emphasis> Shobanov (Diptera,Chironomidae)

Polymorphism and differentiation of the chromosome banding sequence pools and genomic DNA were studied in three natural populations of Chironomus entis from Europe and North America. These populations showed a moderate level of chromosomal polymorphism and high RAPD polymorphism of genomic DNA. The Palearctic and Nearctic populations of this species did not differ significantly in the levels of chromosome and genomic DNA polymorphism. Estimation of the cytogenetic (GD_cg) and genetic (GD_DNA) distances between these C. entis populations showed that their chromosome banding sequence pools and cytogenetic structures are differentiated to a greater extent than genomic DNA. The values of cytogenetic and genetic distances between the Palearctic and Nearctic populations of C. entis are higher than the values of the corresponding distances between the Nearctic populations, but they do not reach the level of divergence between species.     

Genome size and cytogenetic characterization of three Algerian Retama species

Thirty-three populations belonging to the three Retama species, Retama monosperma, Retama raetam and Retama sphaerocarpa, were collected to study species differentiation using flow cytometry for 2C DNA assessment and molecular cytogenetics for karyotype organisation. All were 2n = 48. Genome size ranged from 1.76 to 1.97 pg and revealed significant intraspecific variation correlated to the geographic distribution of the populations. The number and position of the two ribosomal gene families 5S and 45S were determined by fluorescent in situ hybridization, revealing chromosome reorganisation between species. In R. raetam and R. monosperma, the minor 5S loci co-localised with 45S on the satellite chromosome pair. Fluorochrome banding identified GC- and AT-rich DNA regions. In R. monosperma a unique chromomycin positive GC-rich band was observed associated with the secondary constriction. In contrast, an original pattern showing two chromomycin positive bands localised at each side of the extended rDNA locus was observed in R. sphaerocarpa and R. raetam. The polymorphism revealed in our cytogenetic data allowed us to separate the group of R. raetam and R. monosperma from R. sphaerocarpa.     

Karyotypes, chromosome bands and genome size variation in New Zealand endemic gymnosperms

Karyotype studies on 20 taxa of gymnosperms endemic to New Zealand show a wide diversity of chromosome number and form. Fluorochrome banding with DAPI and CMA reveals a depauperate pattern of bands with CMA and no reliable banding with DAPI. Characteristically one pair of chromosomes shows a prominent CMA band, which may or may not be associated with a secondary constriction. A band size polymorphism was observed in all plants ofDacrycarpus dacrydioides, irrespective of the sex of the plant. Measurements of genome size by flow cytometry show a range of values from 12.3 pg to 40.0 pg DNA per 2C nucleus. Intraspecific variation in genome size was observed inManoao colensoi.     

Localization of BAC clones on mitotic chromosomes of <Emphasis Type="Italic">Musa acuminata</Emphasis> using fluorescence <Emphasis Type="Italic">in situ</Emphasis> hybridization

A bacterial artificial chromosome (BAC) library of banana (Musa acuminata) was used to select BAC clones that carry low amounts of repetitive DNA sequences and could be suitable as probes for fluorescence in situ hybridization (FISH) on mitotic metaphase chromosomes. Out of eighty randomly selected BAC clones, only one clone gave a single-locus signal on chromosomes of M. acuminata cv. Calcutta 4. The clone localized on a chromosome pair that carries a cluster of 5S rRNA genes. The remaining BAC clones gave dispersed FISH signals throughout the genome and/or failed to produce any signal. In order to avoid the excessive hybridization of repetitive DNA sequences, we subcloned nineteen BAC clones and selected their ‘low-copy’ subclones. Out of them, one subclone gave specific signal in secondary constriction on one chromosome pair; three subclones were localized into centromeric and peri-centromeric regions of all chromosomes. Other subclones were either localized throughout the banana genome or their use did not result in visible FISH signals. The nucleotide sequence analysis revealed that subclones, which localized on different regions of all chromosomes, contained short fragments of various repetitive DNA sequences. The chromosome-specific BAC clone identified in this work increases the number of useful cytogenetic markers for Musa.     

Chromosome R-banding patterns and NOR homologies in the European wild pig and four breeds of domestic pig

Four European wild pigs and 27 domestic pigs were studied; three Landrace, 12 village pigs from Papua New Guinea, two Chinese pigs Meishan and 10 Creole pigs from the French Antilles. The R-banding patterns were identical for all domestic breeds despite their different history and geographical divergence. The European wild pigs showed a similar R-banding pattern and a centric fusion between pairs 15 and 17 (2n = 36). The nucleolar organizers (NORs) in the European wild pig and the four domestic breeds were localized on the secondary constriction of chromosomes 8 and 10. All animals exhibited in the majority of metaphases two NORs on both chromosomes 10. In some animals. the NORs were expressed only in one of the homologs of chromosome 8. The Chinese pigs had a high amount of silver precipitates on two homologs of chromosome 8. This study confirms several previous reports on the polymorphism of NOR patterns in different domestic pig breeds.     

Nucleoli and ribosomal RNA cistron numbers in Hordeum species and interspecific hybrids exhibiting suppression of secondary constriction

The interspecific hybrids of Hordeum exhibit selective suppression of secondary constriction formation in the chromosome(s) contributed by one of the two parents. A comparison of the number of SAT (secondary constriction) chromosomes in the metaphase cells and the maximum number of nucleoli in interphase cells revealed that the chromosomes capable of organising nucleoli were not always reflected through secondary constriction formation. — The rDNA (DNA complementary to rRNA) amounts were estimated by DNA-rRNA filter hybridisation in diploid and polyploid species of Hordeum and their hybrids. While similar rDNA proportions were present in diploid and autotetraploid lines of H. bulbosum, there were up to threefold differences between H. vulgare and allohexaploids. Furthermore, differences were also apparent between species of same ploidy level. Ribosomal RNA (18S+5.8S+26S) cistron numbers in each of the five experimental hybrids exhibiting the selective suppression of secondary constriction formation revealed no selective loss of rDNA. — The presence of a higher number of nucleoli than the number of SAT chromosomes seen and the presence of expected number of rRNA cistrons suggest that the suppression of secondary constriction formation is not due to selective loss of rDNA.     

The chromosome complement of the Rhesus monkey (Macaca mulatta) determined in kidney cells cultivated in vitro

Summary The chromosome complement of male and female Rhesus monkey has been investigated in kidney cells cultivatedin vitro for 3 to 6 days. The chromosome number is 42. The Y chromosome of the heterogametic male is the smallest element in the complement, and it is acrocentric. The X chromosome ranks eigth in decreasing order of size and typically has an arm ratio of 1.4. The autosomes form a graded size series of metacentric chromosomes, 3–15μ long in early metaphase, and with arm ratios from 1.1 to 3.3. Chromosome IX carries a large secondary constriction near the centromere; it is presumed to be the main nucleolar chromosome. A smaller secondary constriction is found consistently in the long arm of chromosome I. The X chromosome and chromosome XXI appear to be dimorphic in the limited population studied, the alternative forms differing in arm ratios but not in total length. An idiogram of the haploid chromosome complement is presented incorporating measurements of 10 completely analyzed nuclei, five from male monkeys and five from females. On the basis of relative length, arm ratio, and occurrence of secondary constrictions, most chromosomes of the complement can be individually identified. Supported in part by grants from the National Cancer Institute of Canada; the National Institutes of Health of the United States, Public Health Service; and the National Foundation for Infantile Paralysis.     

染色体多态性与临床效应及生殖关系的探究

为了探讨人类染色体结构多态性与生殖异常临床效应的关系,按常规技术方法制备外周血淋巴细胞染色体,经G、C显带,对1 414例遗传咨询者进行核型分析,检出异常核型273例。其中多态性变异180例,占65.93%,非多态性异常核型93例,占34.07％。多态性变异包括D、G组短臂增长10例,次缢痕增长(包括1、9和16号染色体)35例,大Y染色体和小Y染色体 99例,Y染色体臂间倒位6例,9号染色体臂间倒位30例。结果表明,人类染色体多态性与流产、不孕不育、死胎、生育畸形儿等有相关性。     

Cytogenetic variation in farmed stock of Dybowski's sika deer

The chromosome constitution of Dybowski's sika deer was studied on the basis of 15 samples obtained from farmed stock maintained in an enclosure. The diploid chromosome number was 2n=68, 2n=67 and 2n=66. The constitutive heterochromatin (C-bands) was located in the centromeric regions of all acrocentric chromosomes. Metacentric chromosomes were C-negative. Chromosomes of three pairs proved to be NORs carriers. The size polymorphism of silver deposits was identified in two animals. A cytogenetic analysis indicated that the farmed stock of Dybowski's sika deer demonstrates considerable variation. The chromosome polymorphism observed may be a valuable marker for the management and preservation of this species.     

Robertsonian polymorphism and constitutive heterochromatin distribution in chromosomes of the rainbow trout (Salmo gairdneri).

White-blood-cell culture was used to examine the chromosomes of 53 rainbow trout (Salmo gairdneri) from three locations in the Pacific Northwest of the United States. A Robertsonian chromosome polymorphism is present, resulting in diploid numbers of 60, 59, or 58 in different individuals with 104 chromosome arms. The low level of intraindividual Robertsonian variation, differences in the number of subtelocentric chromosomes between individuals with different chromosome numbers, and frequencies of fish with different chromosome numbers in one population suggest that the interindividual differences are inherited and not somatic. C-banding shows that constitutive heterochromatin is localized near the centromeres and near the secondary constriction one chromosome pair.     

Frequency of chromosome polymorphism in Rattus rattus collected in Japan

This paper deals with a population survey of chromosome polymorphism of Rattus rattus collected in Japan and the results of their test crosses. All the animals had diploid 42 chromosomes, but three chromosome pairs, Nos. 1, 9 and 13, were polymorphic in respect to acro- and subtelocentric chromosomes. Frequency of No. 1 chromosome polymorphism in 453 rats collected in 19 localities showed 343 rats (75.5%) with acrocentric homomorphic pair (A/A), 90 (19.9%) with aerocentric and subtelocentric heteromorphic pair (A/S) and the remaining 20 (4.4%) with subtelocentric homomorphic pair (S/S). All animals collected in northern and northwestern Japan showed only the A/A pair, while those collected in southern and southeastern Japan showed A/A, A/S and S/S polymorphism. The latter group was also classified into 3 populations (east, southeast and south) by the different frequency of the subtelocentric chromosome. Progeny tests revealed that segregation of A/A, A/S and S/S types from F₁ hybrids between various chromosome combinations was not significantly different from the theoretical one. However, the number of animals with A/S pair was always slightly higher than the other two types, while those with S/S pair slightly fewer. Local differences of the chromosome polymorphism in Japan were considered due to the result of migration and selection of the rats with S/S chromosome type.Contribution No. 817 from the National Institute of Genetics, Japan. Supported in part by a grant-in-aid from the Ministry of Education of Japan (Scientific Expedition in 1968, and No. 8801 in 1969).     

Genomic content and new insights on the origin of the B chromosome of the cichlid fish <Emphasis Type="Italic">Astatotilapia latifasciata</Emphasis>

B chromosomes are additional chromosomes widely studied in a diversity of eukaryotic groups, including fungi, plants and animals, but their origin, evolution and possible functions are not clearly understood. To further understand the genomic content and the evolutionary history of B chromosomes, classical and molecular cytogenetic analyses were conducted in the cichlid fish Astatotilapia latifasciata, which harbor 1–2 B chromosomes. Through cytogenetic mapping of several probes, including transposable elements, rRNA genes, a repeated DNA genomic fraction (C ₀ t − 1 DNA), whole genome probes (comparative genomic hybridization), and BAC clones from Oreochromis niloticus, we found similarities between the B chromosome and the 1st chromosome pair and chromosomes harboring rRNA genes. Based on the cytogenetic mapping data, we suggest the B chromosome may have evolved from a small chromosomal fragment followed by the invasion of the proto-B chromosome by several repeated DNA families.     

Molecular-cytogenetic studies of ribosomal genes and heterochromatin reveal conserved genome organization among 11 Quercus species

Genomes of 11 Quercus species were characterized using cytogenetic (Giemsa C-banding, fluorochrome banding), molecular-cytogenetic (fluorescence in situ hybridization, FISH, to ribosomal genes) and molecular (dot-blot for ribosomal gene-copy number assessment) techniques. Ribosomal genes are the first DNA sequences to be physically mapped in oaks, and the copy number of the 18S-5.8S-26 S rRNA genes is estimated for the first time. Oak karyotypes were analysed on the basis of DAPI banding and FISH patterns; five marker chromosomes were found. In addition, chromosomal organization of ribosomal genes with respect to AT- and GC-differentiated heterochromatin was studied. Fluorochrome staining produced very similar CMA/DAPI banding patterns, and the position and number of ribosomal loci were identical for all the species studied. The 18S-5.8S-26 S rRNA genes in oak complements were represented by a major locus at the subterminal secondary constriction (SC) of the only subtelocentric chromosome pair and a minor locus at paracentromeric SC of one metacentric pair. The only 5 S rDNA locus was revealed at the paracentromeric region of the second largest metacentric pair. A striking karyotypic similarity, shown by both fluorochrome banding and FISH patterns, implies close genome relationships among oak species no matter their geographic origin (European or American) or their ecophysiology (deciduous or evergreens). Dot-blot analysis gave preliminary evidence for different copy numbers of 18S-5.8S-26 S rRNA genes in diploid genomes of Q. cerris, Q. ilex, Q. petraea, Q. pubescens and Q. robur (2700, 1300, 2200, 4000 and 2200 copies, respectively) that was correlated with the size polymorphism of the major locus. Received: 26 February 1999 / Accepted: 16 March 1999     

The karyotype of Adenomera diptyx (Boettger 1885) (Anura, Leptodactylidae) from northeastern Argentina

In this work we analyzed the karyotype of five populations of Adenomera diptyx from Argentina after conventional staining, Ag-NOR and C-banding. All specimens presented 2n = 26 and FN = 34. The karyotype was formed by three submetacentric, one metacentric and nine telocentric pairs. Silver staining revealed that the NOR was located on a secondary constriction in pair 7. C- banding evidenced constitutive heterochromatin at the pericentromeric region of all chromosomes. The karyotype of A. diptyx was similar to that of A. hylaedactyla (2n = 26, FN = 34) and different from that of A. andreae (2n = 26, FN = 40) in the fundamental number and secondary constriction position. It also differed from the karyotypes of A. marmorata (2n = 24, FN = 34 and 36) and of A. aff. bokermanni (2n = 23, FN = 34) in diploid number. Until a comprehensive cytogenetic analysis of all the species of the genus is performed, their chromosome evolution will remain poorly understood.     

Molecular cytogenetic characterization of breakpoints involving pericentric inversions of human chromosome 9

Pericentric inversions involving the secondary constriction (qh) region of chromosome 9 are considered to be normal variants. The evolutionary mechanisms and conservation of these inversions via Mendelian fashion have been investigated since the advent of banding techniques. Routine cytogenetic techniques cannot provide the fine characterization necessary to determine the type of genetic material involved in these rearrangements. Therefore, the fluorescence in situ hybridization technique with the human centromere-specific alpha satellite and the beta satellite (D9Z5) and classical satellite (D9Z1) human DNA probes were used to identify the breakpoints of chromosome 9 pericentric inversions. Four unique types of pericentric inversions involving the 9qh region were observed, and the mechanism may be due to breakage and reunion at the proposed breakpoints. They are: type A inversions consist of breakpoints within the alpha and beta satellite DNA regions; type B consist of breakpoints within the beta satellite DNA region and band 9q13; type C involve breakage within the beta and classical satellite DNA regions, and type D have breakpoints within the alpha and classical satellite DNA regions. Obviously, reshuffling of satellite DNA sequences has occurred, which has given rise to a variety of heteromorphisms whose clinical significance remains obscure. Received: 21 December 1995 / Revised: 30 May 1996     

眼斑拟石首鱼重复DNA序列的染色体定位

针对眼斑拟石首鱼Sciaenops ocellatus染色体标记匮乏的问题, 利用荧光原位杂交(FISH)定位了眼斑拟石首鱼的18S rDNA、5S rDNA和端粒序列。结果显示, 眼斑拟石首鱼的核型公式为2n=48t; 仅有1对18S rDNA位点, 位于第1对染色体的次缢痕部位; 有2对5S rDNA位点, FISH信号强度不等, 强信号位于第8对染色体的近着丝粒端, 弱信号位于第3对染色体的臂间。端粒FISH信号出现于所有染色体的两端, 但表现出染色体两端信号不平衡的特点, 着丝粒端FISH信号明显强于远端信号。这一特点为判定染色体的方向提供了便利。结合其他石首鱼的核型数据可以推断, 2n=48t的核型及单对近着丝粒分布的18S rDNA位点是石首鱼的共同祖征; 在石首鱼进化过程中, 曾发生活跃但不影响宏观核型的小规模重排。研究结果丰富了眼斑拟石首鱼染色体的辨识标记, 并为研究石首鱼染色体进化提供了基础数据。