中国五种高山锄足蟾的核型、Ag-NORs和C-带的研究

作者用核型、Ag-NORs和C-带,对分布于川、滇两省的二属(齿突蟾、齿蟾)五种(胸腺齿突蟾、圆疣齿突蟾、凉北齿蟾、秉志齿蟾、疣刺齿蟾)锄足蟾作了属间和种间关系的比较分析,并讨论了它们的核型演化机制。结果表明:(1)齿突蟾和齿蟾两属间在核型和带型上都有明显的差异,演化途径主要的可能是含有重复DNA染色体片段的相互易位或臂间倒位;(2)属内不同种之间带型无显著差异,但某些对应染色体对间,其相对长度和臂比值差异明显,十分可能是常染色质片段的易位和臂间倒位所致;(3)凉北齿蟾有染色体数目变异多态现象;(4)五种锄足蟾均未发现异形性染色体。     

Chromosomal analysis of twelve species of Microhylidae (Anura) from Madagascar

The karyotypes of four species of Dyscophinae and eight species of Cophylinae were analyzed. The chromosome number was 2n=26 in all cases. Between the two subfamilies a difference in the form of the karyotype was observed; the chromosomes show a gradual decrease in length in the Dyscophinae, whereas in the Cophylinae the karyotype demonstrates a clear discontinuity of size between pairs 5 and 6.Chromosomal polymorphism was found in Plethodontohyla tuberata, the chromosomes of pair 4 were subtelocentric in the homozygous specimens, whereas this pair showed a subtelocentric and a submetacentric chromosome of equal length in the heterozygous one, suggesting a pericentric inversion. Although in the Cophylinae the chromosome number is constant, the number of chromosome arms is variable. Pericentric inversions seem to play an important role in the chromosomal evolution of the Cophylinae.     

Chromosomal polymorphism of mandarin vole,Microtus mandarinus (Rodentia)

The mitotic and meiotic chromosomes of mandarin vole, Microtus mandarinus Milne-Edwards, from Shandong Province of China were analyzed by conventional, G- and C-banding and Silver-staining techniques. We detected chromosomal polymorphism in the vole, exhibiting diploid chromosome numbers 2n = 48-50 and variable morphology of the 1st pair, one medium sized telocentric pair and the X chromosomes. Four types of karyotypes were revealed in the population. According to banding analysis, there were pericentric inversion, Robertsonian fusion and translocation in M. mandarinus karyotype evolution. The X displayed two different morphologies, which could be explained by prericentric inversion and a telocentric autosome translocation.     

Molecular Definition of Pericentric Inversion Breakpoints Occurring during the Evolution of Humans and Chimpanzees

High-resolution G-banding analysis has demonstrated remarkable morphological conservation of the chromosomes of the Hominidae family members (humans, chimpanzees, gorillas, and orangutans), with the most notable differences between the genomes appearing as changes in heterochromatin distribution and pericentric inversions. Pericentric inversions may have been important for the establishment of reproductive isolation and speciation of the hominoids as they diverged from a common ancestor. Here the previously published primate karyotype comparisons, coupled with the resources of the Human Genome Project, have been used to identify pericentric inversion breakpoints seen when comparing the human karyotype to that of chimpanzee. Yeast artificial chromosome (YAC) clones were used to detect, by fluorescencein situhybridization, five evolutionary pericentric inversion breakpoints present on the chimpanzee chromosome equivalents of human chromosomes 4, 9, and 12. In addition, two YACs from human 12p that detect a breakpoint in chimpanzee detect a similar rearrangement in gorilla.     

Diversified chromosomal characteristics in Centropyge fishes (Pomacanthidae, Perciformes)

We studied karyotypes and other chromosomal markers such as C-banded heterochromatin and Ag-stained nucleolus organizer regions (Ag-NORs), in seven Centropyge fishes (Pomacanthidae, Perciformes). These results revealed diversified chromosomal characteristics in Centropyge species. Three species had 2n = 48 chromosomes, whereas four species had 2n = 52 chromosomes. Fundamental numbers showed a large variation from 48 to 82, particularly in the species with 2n = 52 chromosomes. In all the species, Ag-NORs were located in a single chromosome pair and C-bands were mainly distributed in the centromeric regions of most chromosomes, as commonly seen in teleostean fishes. However, these chromosomal markers showed species-specific variations and provided us with useful information that could help us in understanding chromosomal evolution. On the basis of these chromosomal characteristics, we infer the process of chromosomal evolution, which according to us involves an increase in chromosome number from 2n = 48 to 2n = 52 through centric fission or other mechanisms, and in fundamental number through pericentric inversion. In particular, karyotypic evolution involving the increase in chromosome number is an unusual event in the evolution of higher teleostean groups. Handling editor: K. Martens     

Constitutive heterochromatin, 5S and 18S rDNA genes in Apareiodon sp. (Characiformes, Parodontidae) with a ZZ/ZW sex chromosome system

Karyotype, sex chromosome system and cytogenetics characteristics of an unidentified species of the genus Apareiodon originating from Piquiri River (Paraná State, Brazil) were investigated using differential staining techniques (C-banding and Ag-staining) and fluorescent in situ hybridization (FISH) with 5S and 18S rDNA probes. The diploid chromosome number was 2n = 54 with 25 pairs of meta- (m) to submetacentric (sm) and 2 pairs of subtelocentric (st) chromosomes. The major ribosomal rDNA sites as revealed by Ag-staining and FISH with 18S rDNA probe were found in distal region of longer arm of st chromosome pair 26, while minor 5S sites were observed in the interstitial sites on chromosome pairs 2 (smaller cluster) and 7 (larger one). The C-positive heterochromatin had pericentromeric and telomeric distribution. The heteromorphic sex chromosome system consisted of male ZZ (pair 21) and female middle-sized m/st Z/W chromosomes. The pericentric inversion of heterochromatinized short arm of ancestral Z followed by multiplication of heterochromatin segments is hypothesized for origin of W chromosome. The observed karyotype and chromosomal markers corresponded to those found in other species of the genus.     

两种髭蟾的Ag—NORs,C—带及核型的研究

本文叙述和比较了峨眉髭蟾和哀牢髭蟾的核型,C-带和Ag-NORs,结果表明两者有下列相同方面:2n=26(6+7)、除No.3为SM外,其余诸对概为M,次缢痕和Ag-NoRs位于6q,并都表现出异形现象,No.1长臂有长度异形,但所增染色体片断不呈现C-带正染,C-带正染主要是在各对染色体的着丝点区域,另外No.2短臂近着丝区域亦为正染。这些表明二者之间有很大的核带型同源性。但是二者间在核型的某些对应染色体之间,在相对长度(6对)和臂比值(2对)方面有显著性差异,故可推测其机制是相互易位和臂间倒位。另外,哀牢髭蟾未发现与性别相关的异形性染色体。     

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.     

The chromosomes of Nycticebus coucang (Boddaert, 1785) (Primates: Prosimii)

The high-quality karyotype of a specimen of Nycticebus coucang is described and illustrated. The X chromosome is found to be indistinguishable from that of the greater galagos, and may represent a synapomorphic trait. The Y chromosome is a medium to small submetacentric (3.2% TCL) and constitutes one of the larger Y chromosomes known in primates. N. coucang is found to have multiple NOR-bearing chromosomes in contrast to the single pair found in galagine and catarrhine monkeys. Since a single NOR-bearing pair is often considered ancestral for primates, this new finding may have important implications for the evolution of these cistrons. One of the chromosomal polymorphisms in this specimen is a pericentric inversion, involving a NOR-bearing autosomal pair (no. 6), that alters the position of the active site. Further, homologues 2p differ by aparacentric inversion. These results confirm that lorisiforms are characterized by considerable chromosomal polymorphism.     

Molecular cytogenetics and characterization of a ZZ/ZW sex chromosome system in Triportheus nematurus (Characiformes, Characidae)

Chromosomes of Triportheus nematurus, a fish species from family Characidae, were analyzed in order to establish the conventional karyotype, location of C-band positive heterochromatin, Ag-NORs, GC- and AT-rich sites, and mapping of 18S and 5S rDNA with fluorescence in situ hybridization (FISH). The diploid number found was 2n = 52 chromosomes in both males and females. However, the females presented a pair of differentiated heteromorphic chromosomes, characterizing a ZZ/ZW sex chromosome system. The Z chromosome was metacentric and the largest one in the karyotype, bearing C-positive heterochromatin at pericentromeric and telomeric regions. The W chromosome was middle-sized submetacentric, appearing mostly heterochromatic after C-banding and presenting heterogeneous heterochromatin composed of GC- and AT-rich regions revealed by fluorochrome staining. Ag-NORs were also GC-rich and surrounded by heterochromatic regions, being located at the secondary constriction on the short arms of the second chromosome pair, in agreement with 18S rDNA sites detected with FISH. The 18S and 5S rDNA were aligned in tandem, representing an uncommon situation in fishes. The results obtained reinforce the basal condition of the ZZ/ZW sex system in the genus Triportheus, probably arisen prior to speciation in the group.     

云南三种齿蟾的细胞遗传学研究(锄足蟾科Pelobatidae,无尾目Anura)

本文报道了分布在云南的3种齿蟾(乡城齿蟾、景东齿蟾和棘疣齿蟾)的核型、Ag-NORs和C-带,结果表明2n=26(6+7).NF=52,一对次缢痕和Ag-NORs位于6 q,该区域同时被C-带正染,其余C-带位于各对染色体的着丝点区域。这些特征表现出3种齿蟾的核型同源性和原始性。齿蟾属内种间各对应染色体对在相对长度和臂比值的差异显著性测定表明其核型演化途径可能是臂间倒位和相互易位。3种齿蟾具有染色体数目和Ag-NORs的变异和异形现象,其中乡城齿蟾的一个多余染色体呈现C-带正染,因此可能是B染色体,是锄足蟾科中首次报道。三种齿蟾均未发现与性别相关的异形染色体。     

Cytogenetic comparison of Podocnemis expansa and Podocnemis unifilis: A case of inversion and duplication involving constitutive heterochromatin

Podocnemis expansa and P. unifilis present 2n = 28 chromosomes, a diploid number similar to those observed in other species of the genus. The aim of this study was to characterize these two species using conventional staining and differential CBG-, GTG and Ag-NOR banding. We analyzed specimens of P. expansa and P. unifilis from the state of Tocantins (Brazil), in which we found a 2n = 28 and karyotypes differing in the morphology of the 13^th pair, which was submetacentric in P. expansa and telocentric in P. unifilis. The CBG-banding patterns revealed a heterochromatic block in the short arm of pair 13 of P. expansa and an interstitial one in pair 13 of P. unifilis, suggesting a pericentric inversion. Pair 14 of P. unifilis showed an insterstitial band in the long arm that was absent in P. expansa, suggesting a duplication in this region. Ag-NORs were observed in the first chromosome pair of both species and was associated to a secondary constriction and heterochromatic blocks.     

Karyology and karyotype analysis of diploid freshwater planarian populations of the Dugesia gonocephala group (Platyhelminthes,Tricladida) found in Sardinia

Karyology and karyotype analysis were carried out on freshwater planarian populations of the Dugesia gonocephala group. The strains studied were all diploid with chromosomic number 2n = 16; n = 8. They came from 12 sites mainly localized on the west of the island of Sardinia. Three karyotypes indicated with the letters A, B and C were found in which eight homomorphic pairs of chromosomes were easily identified. In karyotype A all chromosomes are metacentric. Ten populations of the twelve examined showed this karyotype which appears to be the most common. In karyotype B the seventh pair of chromosomes is submetacentric. This karyotype is quite common having been previously found in another eight Sardinian localities. Karyotype C differs from the others in having submetacentric third and seventh pairs of the chromosome complement. It was found in only one locality. The differences observed between these three karyotypes could be interpreted either as sign of differentiation at species level, or as an intraspecific variation due to chromosome mutations (pericentric inversions). This revised version was published online in July 2006 with corrections to the Cover Date.     

Karyotype evolution by pericentric inversion as a stochastic process

We have shown statistically that karyotype change due to pericentric inversion is a Markov process. This implies that acrocentrics (A) are apt to be changed into meta-, submeta- and subtelocentrics (M, SM and ST) by pericentric inversion, and that M, SM and ST chromosomes highly tend to be changed again into M, SM and ST by the rearrangement, i.e. that pericentric inversion is statistically “directional”. This conclusion is consistent with the karyotype data covering 723 mammalian species.     

Studies on the chromosomes of genusNycticebus

The karyotypes of three species (N. coucang, N. intermedius, andN. pygmaeus) of genusNycticebus, collected from the southern Yunnan of China, have been studied. All individuals from three species possess 2n=50 chromosomes, and all chromosomes in their complement are biarm chromosomes. The karyotype of slow loris (N. coucang) is characterized by having a secondary constriction and Ag-NORs in the short arms of pair No. 1. The G-banding patterns of three species are very similar. Three species are found to have multiple Ag-NORs. InN. coucang, NORs were observed on five pairs (Nos. 1, 6, 9, 15, and 23) and inN. intermedius andN. pygmaeus, NORs were found on four pairs (Nos. 6, 9, 15, and 20). This finding indicates that slow lorises, as primitive primates, also have multiple NOR-bearing chromosomes. Finally, the classification of genusNycticebus by karyotype analysis is discussed, and our results suggest that there are at least two valid species, namely:N. coucang andN. pygmaeus.     

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).     

Similarity of giemsa banding patterns of chromosomes in several species of the Genus Rattus

Giemsa banding patterns of chromosomes in seven Rattus species were compared. Four species (R. rattus tanezumi, R. norvegicus, R. exulans and R. muelleri) had all 2n=42 and their karyotypes and banding patterns were similar, although slight differences were observed. Another subspecies (R. rattus rattus) and two other species (R. fuscipes and R. conatus) had fewer chromosomes than the above species by having large biarmed chromosomes developed probably by Robertsonian fusion. The origin of the arms of biarmed chromosomes was recognized by their characteristic banding patterns. The remaining species, R. sabanus, had a karyotype markedly different from the other species by having two small metacentrics although in the others their number was 7. Banding patterns of the chromosomes in this species, however, were also very similar to those of the other, and therefore the 7 small metacentrics seemed to have originated by pericentric inversion of small acrocentrics.Contribution No. 912 from the National Institute of Genetics, Japan. Supported by a grant-in-aid from the Ministry of Education of Japan, Nos. 90183, 90375 and 744002.     

Molecular characterization of the pericentric inversion of chimpanzee chromosome 11 homologous to human chromosome 9

In addition to the fusion of human chromosome 2, nine pericentric inversions are the most conspicuous karyotype differences between humans and chimpanzees. In this study we identified the breakpoint regions of the pericentric inversion of chimpanzee chromosome 11 (PTR 11) homologous to human chromosome 9 (HSA 9). The break in homology between PTR 11p and HSA 9p12 maps to pericentromeric segmental duplications, whereas the breakpoint region orthologous to 9q21.33 is located in intergenic single-copy sequences. Close to the inversion breakpoint in PTR 11q, large blocks of alpha satellites are located, which indicate the presence of the centromere. Since G-banding analysis and the comparative BAC analyses performed in this study imply that the inversion breaks occurred in the region homologous to HSA 9q21.33 and 9p12, but not within the centromere, the structure of PTR 11 cannot be explained by a single pericentric inversion. In addition to this pericentric inversion of PTR 11, further events like centromere repositioning or a second smaller inversion must be assumed to explain the structure of PTR 11 compared with HSA 9.     

颈棱蛇核型和Ag－NORs的研究

本文研究了云南产颈棱蛇的核型和Ａｇ－ＮＯＲｓ。其Ｚｎ＝４６,由８对大染色体和１５对小染色体组成,大染色体中Ｎｏ．６、８为亚中着丝粒染色体,其余均为端或亚端着丝粒染色体。在Ｎｏ．２的末端有一明显的次缢痕,常常只在一条同源染色体上可见。第４对大型染色体为性染色体,雌性为ＺＷ型,其中Ｚ染色体较大,而Ｗ染色体长度仅为Ｚ染色体的三分之二。一对核仁组织者（ＮＯＲｓ）位于Ｎｏ．２末端,其位置与次缢痕位置相对应。文中还对颈棱蛇的分类和进化进行了讨论。     

Synteny comparison between apes and human using fine-mapping of the genome

Comparing the genomes of the great apes and human should provide novel information concerning the origins of humankind. Relative to the great apes, the human karyotype has one fewer chromosome pair, as human chromosome 2 derived from the telomeric fusion of two ancestral primate chromosomes. To identify the genomic rearrangements that accompanied human speciation, we initiated a comparative study between human, chimpanzee, and gorilla. Using the HAPPY mapping method, an acellular adaptation of the radiation hybrid method, we mapped a few hundred markers on the human, chimpanzee, and gorilla genomes. This allowed us to identify several chromosome rearrangements, in particular a pericentric inversion and a translocation. We precisely localized the synteny breakpoint that led to the formation of human chromosome 2. This breakpoint was confirmed by FISH mapping.