Relationship between the ratio of parental chromosomes and parental doubling times in Chinese hamster-mouse somatic cell hybrids

Somatic hybrids obtained by the selective method of Littlefield between a permanent line of Chinese hamster cells (Wg3) and one of mouse cells (3TP) showed a preponderance of biarmed (hamster) chromosomes. Under normal culture conditions (37°) the doubling time of the parental mouse cells was twice as long as that of the parental hamster cells. If the temperature of incubation was lowered (31°), the relative difference in doubling times was reduced; in hybrid lines obtained under these conditions, the proportion of biarmed chromosomes was also reduced. Upon extended cultivation the average number of telocentric chromosomes progressively decreased in all hybrid lines tested, regardless whether these were started and maintained at 37° or at 31°. An inverse correlation was observed in hybrid cells between doubling time and relative proportion of biarmed chromosomes, suggesting that the karyotypic changes observed after extended culture were due to the selective overgrowth of cells with a high biarmed to telocentric ratio.     

Chromosome markers of murine hybridomas

The karyological study of 10 mouse hybridomas revealed that all cells in two hybridoma clones, as well as in two subclones isolated from the third hybridoma, contained specific clonal biarmed markers, atypical for myeloma parent cells X63.Ag8.653. The proportion of cells with additional new meta- and submetacentric markers, which were different in the cells of the same culture, reached 0.38-0.56 in some of the hybridomas under study. The above biarmed chromosomes were, probably, formed as the result of the centromeric fusions of subtelocentrics. The presence of identical new biarmed chromosomes in all cells in some hybridoma cultures could be attributed to the fact that all these cells originated from a single initial cell, already containing such marker (or markers). The results of the cytogenetic analysis may confirm the monoclonal origin of a considerable part of mouse hybridomas.     

Establishment of the AU-bek cell line and comparison with two other bovine cell lines

Summary Establishment of a new bovine cell line, AU-BEK, is reported. The cell line developed in a culture initiated from bovine embryonic kidneys by spontaneous cultural alteration to epithelioid cells that are indefinitely propagable. Epithelioid cells gradually increased to become the predominant cell. Whereas normal bovine cells have a diploid number of 60 chromosomes, of which only the two sex chromosomes are biarmed, AU-BEK cells at the 80th passage had a modal chromosome number of 84 and an average of 30 biarmed chromosomes per cell. AU-BEK cells are now in their 220th passage. Of the AU-BEK, MDBK, and CKT-1 bovine cell lines, the CKT-1 cell line had a karyotype closest to that of normal bovine cells. Their modal chromosome number was 57, and only three biarmed chromosomes were usually present. The bovine character of AU-BEK and CKT-1 cells was established by cytotoxic and viral susceptibility tests. Supported by the Alabama Agricultural Experiment Station. Publication No. 1115, School of Veterinary Medicine, Auburn University.     

Polymorphism of the Sphaerium corneum (Bivalvia, Veneroida, Sphaeriidae) revealed by cytogenetic and sequence comparison

Two populations of Sphaerium corneum were sampled from River Vilnelė and small pond in Vilnius, Lithuania. The chromosomes were studied using conventional Giemsa staining and karyometric analysis. Inter- and intra-individual variation in the diploid chromosome numbers was revealed and two different sources of chromosome variability were identified: B chromosomes and the structural changes of chromosomes of the basic (A) set. The chromosome set of the more common karyotypic form, 2 n  = 30, found in both populations, consists of all biarmed metacentric and meta-submetacentric chromosomes of gradually decreasing size. Small, biarmed, mitotically unstable B chromosomes were found in the cells of this karyotypic form. Specimens with 2 n  = 36 were found only in pond. No B chromosomes were detected in their cells. The karyotype is characterized by presence of two pairs of medium telocentrics and four pairs of small subtelocentrics. The remaining chromosomes are biarmed. Robertsonian fusions appear to be involved in formation of two karyotypic forms of S. corneum . DNA sequence analyses showed that ITS1 is identical in both karyotypic forms. On the other hand, differences in 16S sequence were revealed and two haplotypes, corresponding to two karyotypic forms, were identified. The present study opens new perspectives in establishing species-specific characters for confident identification of Sphaerium species and provides insights to the genetic intraspecific variability and possible mechanisms of speciation.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 53–64.     

Biotope distribution of cestodes from the common shrew Sorex araneus in southern Karelia

Specific features in the formation of cestode species composition in the common shrew in different biotopes have been analyzed. Four categories of parasites have been recognized depending on a degree of dominance: dominants (Neoskrjabinolepis singularis, Molluscotaenia crassiscolex), subdominants (Ditestolepis diaphana), adominants A (Staphylocystis furcata, Vigisolepis spinulosa, adominants B (Hymenolepis scutigera, Dilepis undula, D. secunda, Taenia mustelae). A significant similarity was discovered between the species diversity and the dominance characteristics of cestodes. It was found that the distribution of mass species of parasites in the host population corresponds to the gamma distribution model. In regard to cestodes examined, a hypothesis considering the spatial structure and size of territory occupied by stable parasitic system is put forward.     

Karyotype of a Japanese salamander Hynobius katoi and its implication on breeding ecology (Amphibia: Caudata)

Karyotype of a Japanese small salamander, Hynobius katoi, was first described. All individuals examined had 2n=58 chromosomes, consisting of nine pairs of biarmed macrochromosomes, four pairs of biarmed medium-sized chromosomes, six pairs of biarmed microchromosomes, and 10 pairs of uniarmed microchromosomes, although distinction of the second and the third groups of chromosome pairs was not clear. All pairs appeared homologous and no sexual dimorphism was found. Possession of 2n=58 chromosomes in H. katoi strongly suggests its lotic-breeding habits as was expected from the number and size of eggs and adult morphology. When compared morphology of chromosomes among lotic-breeders with 2n=58 chromosomes, metacentric nature of No. 10 seems to characterize the karyotype of H. katoi.     

Chromosome conservation in the Bovidae.

The chromosomes of 12 bovid species were harvested from fibroblast cultures after incorporation of bromodeoxyuridine into early replicating DNA. Q-band karyotypes were constructed, and, when possible, autosomal arms were numbered according to the cattle standard karyotype. Diploid chromosome number ranged from 30 to 60, yet, based on band similarity, chromosome-arm homologies were extensive. Employing the cattle karyotype as the standard, autosomal-arm differences indicative of possible syntenic disruption were noted for only chromosomes 3, 9, and 14. While chromosome-arm homologies were extensive, shared homologous biarmed chromosomes were rare. The commonness of monobrachially homologous biarmed chromosomes among some bovids (e.g., Antilopinae) suggested that reproductive isolation and speciation in some instances might have resulted from centric fusion events.     

The karyotypes of Triaenophorus nodulosus and T. crassus (Cestoda: Pseudophyllidea)

Two species of the genus Triaenophorus were found to have widely different chromosome sets. The karyotype of T. nodulosus consists of 26 biarmed chromosomes ranging from 1.45 to 6.75 microns long. The diploid set of T. crassus contains 18 chromosomes with a well-distinguished first pair of large metacentric homologues. All the chromosomes with the exception of the last pair of acrocentric elements are biarmed. Their absolute length ranges from 1.50 to 8.50 microns. The possible pathways of karyotype differentiation and the evolution of these species are discussed.     

How many times has polyploidization occurred during acipenserid evolution? New data on the karyotypes of sturgeons (Acipenseridae,Actinopterygii) from the Russian Far East

The karyology of species of sturgeon from the Russian Far East demonstrates that the karyotype of the Sakhalin sturgeon (Acipenser mikadoi) includes 262 ± 4 chromosomes with 80 biarmed chromosomes and the number of chromosome arms (NF) 342 ± 4, the karyotype of the Amur sturgeon (A. schrenckii) includes 266 ± 4 chromosomes with 92 biarmed chromosomes and NF 358 ± 4, and the karyotype of the kaluga (A. dauricus) consists of 268 ± 4 chromosomes with 100 biarmed chromosomes and NF 368 ± 4. These results prove that all western Pacific sturgeon species are from a tetraploid origin, based on a recent ploidy scale. This suggests that at least three polyploidization events have occurred during the evolution of Acipenseridae. However, if polyploid species originated by hybridization between diploid species, there may have been more polyploidization events in this group of fishes.     

Cytogenetic comparison of saola (Pseudoryx nghetinhensis) and cattle (Bos taurus) using G- and Q-banding and FISH

In order to cytogenetically describe the new bovid species saola (Pseudoryx nghetinhensis), comparative G- and Q-banding of saola and cattle (Bos taurus) chromosomes as well as FISH-mapping of 32 type-I markers (29 Texas markers and three additional markers) on saola chromosomes were performed. Saola was shown to have a diploid number of 2n = 50 chromosomes possessing five biarmed autosomal pairs and an acrocentric X chromosome. Homology of saola and cattle chromosomes was indicated by banding patterns and by marker hybridization suggesting that all five biarmed pairs in saola originate from centric fusions involving ten cattle autosomes. However, small intrachromosomal rearrangements cannot be excluded. In this study the first preliminary homology map of these two species is presented.     

Adriamycin-induced centric fusions in mouse chromosomes: in vivo and in vitro analysis

The nature of metacentric chromosomes induced by the anthracycline antibiotic adriamycin (ADR) in mice was studied. The analysis of bone marrow cells of mice injected intraperitoneally with a single dose of the chemical and sacrificed at different posttreatment lapses demonstrated the persistence of biarmed chromosomes even after 30 days. Observations of BUdR-substituted chromosomes from primary cell cultures after one replication cycle revealed the maintenance of DNA polarity through the centromere in most of the metacentric chromosomes. These facts could be considered as a good indication that ADR induces Robert-sonian fusions in mouse chromosomes.     

Centromere organization in chromosomes of the mouse

The ultrastructure of the centromere region of chromosomes from mouse L929 cells treated with agents that affect centromere condensation have been examined using light, transmission electron, and scanning electron microscopic techniques. Micrographs of expanded centromeres from treated chromosomes illustrate that both the biarmed chromosomes that were generated by Robertsonian fusion during the past history of the strain and the functional centromere of the multicentromeric marker chromosomes display a prominent gap. This gap probably represents the original site of association of the acrocentric chromosomes and is also the site of the kinetochore. Despite the multicentromeric nature of the marker chromosome a single pair of kinetochores were found only at the central heterochromatic region. The functional implications of these structural findings are discussed.     

Human-Mouse Hybrid Cell Lines and Susceptibility to Polio Virus : II. Polio Sensitivity and the Chromosome Constitution of the Hybrids

A number of human-mouse hybrid cell lines with partial human chromosome complements were sensitive to poliovirus because the cells contained the viral receptor substance of human origin. Infection of the lines with one type of poliovirus regularly led to the survival of a few cells, whose progeny were found to be resistant to all types of poliovirus. Comparison of the chromosomes of sensitive hybrids and their resistant sublines showed no consistent difference in the number of biarmed human chromosomes of any group. The number of acrocentrics was always lower in the resistant hybrids than in the corresponding sensitive lines. It is suggested that the human chromosome bearing the polio receptor gene is an acrocentric.     

Induction of pigmentation by continuous X-irradiation of amelanotic tumors of B16-XI mouse melanoma and induced change in chromosomes of amelanotic cells

Non-pigmented tumor cells of B16-XI mouse melanoma were found to contain a diploid number of chromosomes similarly to those of melanotic tumors and the parental cells in tissue culture. A major difference between pigmented and non-pigmented cells was in the number of biarmed chromosomes per cell. There was no difference in growth rate between non-pigmented and pigmented tumors, but growth usually begins about 2 days earlier in the former. Pigmentation lost in the course of serial transplantation was restored by irradiating the non-pigmented tumor continuously with 2,500-3,000 rads/passage of X-rays during six transfer generations. In the course of repeated irradiations, the chromosomes changed structurally and numerically as the pigmentation of the tumor was gradually restored. The observations of tumor growth and chromosomal changes are discussed in relation to the pigmentation of B16-XI melanoma cells.     

Chromosomal evolution in the Brazilian lizards of genus Leposoma (Squamata, Gymnophthalmidae) from Amazon and Atlantic rain forests: banding patterns and FISH of telomeric sequences

An extensive karyotype differentiation was found among three species of gymnophthalmid lizard genus Leposoma which occur in the tropical forest areas of Brazil. We examined the chromosomes of the Amazonic species L. guianense (LOU) and L. oswaldoi (LOS) and the Atlantic forest species L. scincoides (LSC) after conventional and differential staining, and FISH of telomeric sequences. Both Amazonic species shared very similar 2n = 44 karyotypes, including 20 biarmed macrochromosomes and 24 microchromosomes (20 M + 24 m). However, the location of Ag-NORs and the amount of constitutive heterochromatin differed in these karyotypes. The Atlantic forest species L. scincoides has a very distinct karyotype with 52 acrocentric and subtelocentric chromosomes of decreasing size. Comparative R-banding analysis revealed complete homeology of the macrochromosomes of LGU and LOS and correspondence of banding patterns between LSC acrocentrics and subtelocentrics and some arms of biarmed LGU and LOS chromosomes. Pair 1 had similar banding patterns in the three species, implying the occurrence of a pericentric inversion. Interstitial telomeric bands (ITBs) detected by FISH at the pericentromeric region of some biarmed LGU and LOS chromosomes could be remnants of chromosomal rearrangements occurred during the differentiation of the karyotypes. Robertsonian rearrangements as well as pericentric inversions events probable were involved in the karyotype evolution of these Amazon and Atlantic forests species of Leposoma.     

Ag-NOR staining and in situ hybridization of rDNA in the chromosomes of the South American camelids

The location and frequency of Ag-stained NORs and sites of rDNA hybridization were studied in the chromosomes of the South American camelids. In the four camelids these regions occur distally on chromosomes 18, 21, and 27 and the smallest biarmed elements. Quantitative analysis of NOR distribution showed variations between both cells and species. In llama, guanaco and alpaca the NORs number averaged 6 per cell, this being higher than in vicuña where the average was 3. Relative frequencies of NOR-bearing chromosomes in the four camelids were similar. Yet, in vicuña the virtual absence of NOR sites on one of the smallest biarmed pairs was observed. The rDNA sites assessed in llama and vicuña by in situ hybridization with cloned 18S DNA were coincident with the NOR locations and with the frequencies characteristics for each species. Moreover, varying the exposure time of the autoradiographs, labeling patterns specific for each camelid were observed. Grain counts on individual chromosomes indicated that under our conditions one month exposure is enough to demonstrate all the rDNA sites available in the complement of llama. Conversely, at least two months are necessary to show the total sites existing in vicuña. Most probably this finding reflects the presence of variations in the amount of copies of the ribosomal genes per chromosome.     

Evolutionary conservatism in arrangement of genetic material

The diploid number of the Rhesus macaque, Macaca mulatta, is 42. All chromosomes are biarmed and all constitutive heterochromatins are centromeric. The diploid number of the African Green monkey, Cercopithecus aethiops, is 60. Again all chromosomes are biarmed, but seven pairs possess very short second arms which are heterochromatic. The heterochromatins of remaining chromosomes are centromeric. Using G-banding and deleting the heterochromatic short arms, the chromosomes of the African Green monkey can be artificially fused to reconstruct a karyotype of the Rhesus with only one pair of unmatched small metacentrics. In addition to the Robertsonian type of translocations, several sets of centromere-telomere translocations were found. The latter type of translocation reduced three arms into two. Thus the fundamental number can be changed by two mechanisms: growing extra heterochromatic arms and the centromere-telomere fusions.     

A comparative chromosome study of twenty killifish species of the genus Fundulus (Teleostei: Cyprinodontidae)

Female karyotypes from ovarian cell cultures of 20 species of killifish (Fundulus) ranged in diploid number from 32 to 48, but in arm number (NF) from 48 to 52. The small F chromosomes, which constituted the fundamental elements in the karyotype, were evenly graded in length. The large biarmed chromosomes (L), which were about twice the length of the average Fs, characterized only those species with 2N less than 48 chromosomes. And among these species, an increase in complement by a pair of L's was always accompanied by a decrease of two pairs of A's, indicating Robertsonian changes by the centric fusion of two A's to form one L chromosome. Other diagnostic chromosome characters included: the number and structure of biarmed and satellited F chromosomes and the percentage of F's with relatively short short-arms (SSA). Besides centric fusion, mechanisms of chromosomal evolution in Fundulus probably included pericentric inversion, producing biarmed F chromosomes from acrocentric F's and partial loss of a chromosome segment producing smaller biarmed F chromosomes from larger ones. The percentage of SSA chromosomes generally decreases from relatively primitive to specialized species. The presumably most primitive species have only SSA type acrocentric F chromosomes. The 20 Fundulus species were classified into 2 major groups according to the percentage of SSA chromosomes: the SSA group, including 3 subgroups, had more than 50% SSA's; the LSA group, including 2 subgroups, had fewer than 50% SSA's. This classification based only on karyotypic characters generally agreed with others based on gross morphological characters. A possible evolutionary scheme is proposed to account for the derived killifish karyotypes.     

Karyotypes of Rana tagoi Okada with diploid number 28 in the Chausu Mountains of the Minamishinshu district of Nagano Prefecture, Japan (Anura: Ranidae)

Karyotypes of Tago's brown frog Rana tagoi from the Chausu mountains in Minamishinshu of Nagano Prefecture were examined by conventional Giemsa staining, C-banding and late replication (LR)-banding. Chromosome number was 2n = 28 in all cases. The 28 chromosomes consisted of four pairs (1-4) of large biarmed chromosomes, two pairs (5-6) of telocentric chromosomes and eight pairs (7-14) of small biarmed chromosomes. Chromosome pair 11 had a secondary constriction on the long arm. In females, the C-band on the long arm of chromosome pair 6 was detected in both homologs, but was absent from the arms of the homologs of chromosome pairs 5 and 9. In males, C-bands were found in the long arms of both homologs of chromosome pairs 5 and 6, were present only in one homolog of chromosome pair 5 for certain male specimens and found in only one homolog of chromosome pair 9. Specimens of R. tagoi (2n = 28) should thus have two pairs of telocentric chromosomes to provide the same number of chromosome arms, these originating quite likely from chromosome pair 1 in the 26-chromosome specimens by centric fission. Heteromorphic sex chromosomes of the XX-XY type in R. tagoi (2n = 28) in the Chausu mountains were identified. Karyotypes of tail-tip cells from a hybrid tadpole between female R. tagoi (2n = 26) from the Hinohara village in Tokyo and male R. tagoi (2n = 28) from the Chausu mountain population were examined by squash preparation. Chromosome number was 2n = 27 in all tadpoles. The 27 chromosomes consisted of one chromosome set of R. tagoi (2n = 28) and one of R. tagoi (2n = 26).     

黄鳝体内寄生胃瘤线虫的染色体核型及G-带分析

采用常规空气干燥法制片,对寄生于黄鳝(Monopterus albus)体腔内的胃瘤线虫(Eustrongylidesignotus)染色体核型进行分析。结果表明:胃瘤线虫体细胞有12条染色体,为二倍体,核型公式为2n=12=10 m+2 sm。由5对常染色体和1对性染色体组成,性别决定模式为XX-XY,其中X、Y和1～4号染色体都为中着丝粒染色体,5号为亚中着丝粒染色体。每对染色体都有特定的G-带带型。