Chromosomal location and distribution of As51 satellite DNA in five species of the genus Astyanax (Teleostei, Characidae, Incertae sedis)

Constitutive heterochromatin makes up a substantial portion of the genome of eukaryotes and is composed mainly of satellite DNA repeating sequences in tandem. Some satellite DNAs may have been derived from transposable elements. These repetitive sequences represent a highly dynamic component of rapid evolution in genomes. Among the genus Astyanax , the As51 satellite DNA is found in species that have large distal heterochromatic blocks, which may be considered as derived from a transposable DNA element. In the present study, As51 satellite DNA was mapped through in situ fluorescent hybridization in the chromosomes of five species of the genus. The possible roles of this type of saltatory DNA type in the genome of the species are discussed, along with its use for the phylogenetic grouping of the genus Astyanax , together with other shared chromosomal characters. However, the number of As51 clusters is presented as a homoplastic characteristic, thereby indicating evident genomic diversification of species with this type of DNA.     

The species complex Astyanax fasciatus Cuvier (Teleostei, Characiformes) – a multidisciplinary approach

Cytogenetic data have provided important clues that the Astyanax fasciatus populations from the Upper Paraná River basin could be a part of a more diverse fish group, usually included on the same taxa. Samples collected in Cachoeira de Emas, SP, in Mogi-Guaçu River basin, show two major cytotypes presenting 2 n = 46 and 2 n = 48 chromosomes, with distinct karyotypic formula, despite the fact that the molecular data suggested some degree of gene flow between these cytotypes. Cytogenetic and morphometric analyses were performed in this species, aiming to contribute to the understanding of the natural history from such fish group. Two allopatric populations with distinct standard cytotypes were analysed, and the data obtained suggest the separation into two groups.     

Cytogenetic divergence in two sympatric fish species of the genus Astyanax Baird and Girard, 1854 (Characiformes,Characidae) from northeastern Brazil

The fish genus Astyanax is widespread throughout the Neotropical region and is one of the most species-rich genera of the Characiformes. Cytogenetic studies of Astyanax have revealed marked intra- and interspecific diversity, with the identification of various species complexes. In this report, we describe the karyotypic structure of two sympatric species of Astyanax (Astyanax sp. and Astyanax aff. fasciatus) from the Middle Contas River basin in the northeastern Brazilian state of Bahia. Both species had 2n = 48 but differed in their karyotypic formulae. Small heterochromatic blocks and multiple nucleolar organizer regions (NORs) were identified in both species. Terminal CMA₃⁺/DAPI⁻ signals were observed in Astyanax sp. and A. aff. fasciatus, mostly coincident with NORs. These results show that chromosomal markers can be used to identify species in this fish complex. These markers can provide useful information for evolutionary studies and investigations on the mechanisms of chromosomal diversity in Astyanax.     

Occurrence of macro B chromosomes in Astyanax scabripinnis paranae (Pisces,Characiformes, Characidae)

B chromosomes occur in several Neotropical fish species. Cytogenetic analysis of 27 specimens (15 females and 12 males) of Astyanax scabripinnis paranae from the Araquá river (a small headwater tributary of the Tietê river) shows that this population has 2n=50 chromosomes (4M+30 SM+4ST+12A), two chromosome pairs with NORs and conspicuous C-band positive blocks in the terminal position of the long arm of four chromosome pairs. In this population, eight females presented 2n=51 chromosomes and the extra chromosome was a large metacentric similar in size and morphology to the first chromosome pair in the karotype. This accessory chromosome is entirely heterochromatic in C-banded metaphases and shows a late replication pattern evidenced by BrdU incorporation. There was no significant correlation between the presence of B chromosomes and increased NOR activity at the P>0.05 level. Some aspects related to these B chromosomes are discussed.     

Comparative cytogenetics in Apareiodon affinis (Pisces, Characiformes) and considerations regarding diversification of the group

Comparative cytogenetic studies in Apareiodon affinis (Pisces, Characiformes) from two hydrographic Brazilian basins showed significant divergences related to the general karyotype structure, C‐banding and nucleolar organizer region (NOR) bearing chromosomes. In the upper Paraná basin population, distinct diploid numbers were observed among sexes, the females showing 2n = 55 and the males 2n = 54 chromosomes, characterizing a multiple sex chromosome system of the ZZ/ZW₁W₂ type. A diploid number equal to 54 chromosomes was found for the Cuiabá river population, without a sex chromosome heteromorphism. However, the occurrence of acrocentric chromosomes represents an unique character for this population. These karyotypic differences indicate that the analyzed populations must represent distinct Apareiodon species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interpopulational comparative cytogenetics analysis among three Astyanax (Characiformes: Incertae Sedis) species of two streams of upper Paraná River basin,Brazil

Populations of Astyanax paranae, Astyanax fasciatus and Astyanax altiparanae collected from the Pindorama and Lopei streams (upper Paraná river basin, Brazil) were cytogenetically studied, checking for chromosomal differentiation and the evolutionary trend in the three species. Forty-eight chromosomes (8m + 20sm + 10st + 10a) were observed in both populations of Astyanax paranae. Astyanax fasciatus exhibited 50 chromosomes (8m + 18sm + 10st + 14a, Pindorama stream; 8m + 14sm + 12st + 16a, Lopei stream). The diploid number for A. altiparanae was 50 chromosomes in both populations (6m + 30sm + 4st + 10a). The three studied species exhibited multiple AgNORs with differences among the populations of A. paranae and A. fasciatus. Regarding the heterochromatin distribution, interpopulational differences were observed in the three species. The data of the present study demonstrate that both A. paranae and A. fasciatus exhibit karyotypical interpopulational differences more conspicuous than A. altiparanae populations, probably due to the characteristics that restricted both the A. paranae and the A. fasciatus to the upper portion of the streams, leading to the fixation of different chromosomal rearrangements in the populations. However, the absence of conspicuos interpopulational karyotypical differences in A. altiparanae suggests a gene flow between both populations. In the present study, the role of the streams as an environment which maintains the diversity of fish with different genetic pools became evident.     

Chromosomal location of 18S and 5S rDNA sites in Triportheus fish species (Characiformes, Characidae)

The location of 18S and 5S rDNA sites was determined in eight species and populations of the fish genus Triportheus by using fluorescent in situ hybridization (FISH). The males and females of all species had 2n = 52 chromosomes and a ZZ/ZW sex chromosome system. A single 18S rDNA site that was roughly equivalent to an Ag-NOR was detected on the short arms of a submetacentric pair in nearly all species, and up to two additional sites were also observed in some species. In addition, another 18S rDNA cluster was identified in a distal region on the long arms of the W chromosome; this finding corroborated previous evidence that this cluster would be a shared feature amongst Triportheus species. In T. angulatus, a heterozygotic paracentric inversion involving the short arms of one homolog of a metacentric pair was associated with NORs. The 5S rDNA sites were located on the short arms of a single submetacentric chromosomal pair, close to the centromeres, except in T. auritus, which had up to ten 5S rDNA sites. The 18S and 5S rDNA sites were co-localized and adjacent on the short arms of a chromosomal pair in two populations of T. nematurus. Although all Triportheus species have a similar karyotypic macrostructure, the results of this work show that in some species ribosomal genes may serve as species-specific markers when used in conjunction with other putatively synapomorphic features.     

Differentiated ZZ/ZW sex chromosomes in Apareiodon ibitiensis (Teleostei,Parodontidae): cytotaxonomy and biogeography

Conventional and molecular chromosomal analyses were carried out on three populations of Apareiodon ibitiensis sampled from the hydrographic basins of the São Francisco River and Upper Paraná River (Brazil). The results reveal a conserved diploid number (2n = 54 chromosomes), a karyotype formula consisting of 50 m‐sm + 4st and a ZZ/ZW sex chromosome system that has not been previously identified for the species. C‐banding analysis with propidium iodide staining revealed centromeric and terminal bands located in the chromosomes of the specimens from the three populations and allowed the identification of heteromorphism of heterochromatin regions in the Z and W chromosomes. The number of 18S sites located through fluorescent in situ hybridization (FISH) varied between the populations of the São Francisco and Upper Paraná Rivers. The location of 5S rDNA sites proved comparable in one pair of metacentric chromosomes. Thus, the present study proposes a ZZ/ZW sex chromosome system for A. ibitiensis among the Parodontidae, and a hypothesis is presented regarding possible W chromosome differentiation stages in this species through DNA accumulation, showing geographical variations for this characteristic, possibly as a consequence of geographical reproductive isolation.     

Cytotaxonomy of Astyanax (Characiformes,Characidae) from the Upper Iguaçu River Basin: confirmation of the occurrence of distinct evolutionary units

Taxonomic studies of the genus Astyanax from the Iguaçu River (Brazil) indicate that they may be differentiated into 11 distinct species, some of which have not been formally described and named so for. This study focuses on three of these species, Astyanax sp. B, Astyanax sp. C and Astyanax sp. D from the Upper Iguaçu River Basin. Comparative cytogenetic analyses of C‐banding, Ag‐NORs (silver nitrate stained nucleolar organizer region) and 18S and 5S rDNA corroborate that they are distinct species. A diploid number of 50 chromosomes and similar karyotypic formulae were observed in the three taxa, with the exception of Astyanax sp. D that differs in the number of submetacentric and subtelocentric chromosomes. However, the NOR silver‐staining pattern, the heterochromatic bands (C‐bands) and the mapping of the 18S and 5S rDNA sites in the chromosomes showed divergences between all three species under study, supporting the occurrence of distinct evolutionary units.     

Molecular cytogenetics of <Emphasis Type="Italic">Oligosarcus hepsetus</Emphasis> (Teleostei,Characiformes) from two Brazilian locations

Karyotype and cytogenetic markers of Oligosarcus hepsetus from two Brazilian locations in the Paraíba do Sul River Basin (Brazil) were investigated using differential staining techniques (C-banding, silver (Ag)- and chromomycin A₃ (CMA₃)-staining) and fluorescent in situ hybridization (FISH) using 18 S rDNA and 5 S rDNA probes. The diploid chromosome number was invariably 2n = 50 with 3 pairs of metacentric, 5 pairs of submetacentric, 8 pairs of subtelocentric and 9 pairs of acrocentric chromosomes. No heteromorphic sex chromosomes were observed. The nucleolar organizer regions (NORs) were detected in the short arms of the largest acrocentric pair using Ag-, CMA₃- stainings and FISH with 18 S rDNA probe, the latter showing also positive labeling in the short arms of a small acrocentric pair, not visualized by the former methods. FISH with 5 S rDNA probe showed positive labeling in the two chromosome pairs. While the CMA₃-staining exhibited GC-rich heterochromatin segments in two pairs of chromosomes, including those coincided with Ag-NORs, the DAPI staining did not reveal any signal, indicating the absence of AT-rich heterochromatin. FISH with an As-51 satellite DNA probe derived from the closely related Astyanax scabripinnis did not reveal any positive signal, demonstrating the absence of this class of DNA in the genome of the specimens under study.     

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.     

Chromosomal diversity in the genus Arvicanthis (Rodentia, Muridae) from East Africa: a taxonomic and phylogenetic evaluation

In this paper we discuss the contribution of cytogenetics to the systematics of Arvicanthis in East Africa, by reviewing all the known chromosomal cytotypes of the genus in the area. We also provide G‐ and C‐banding comparisons for two recently described karyotypes, provisionally named ANI‐5 (2n = 56, NFa = 62) and ANI‐6 (2n = 60, NFa = 72). This, therefore, brings the total number of known cytotypes in this area to 10. Five of these correspond to the species recognized by the latest rodent checklist, i.e. A. nairobae (2n = 62, NFa = 78), A. neumanni (2n = 52–53, NFa = 62), A. blicki (2n = 48, NFa = 62), A. abyssinicus (2n = 62, NFa = 64) and A. niloticus (2n = 62, NFa = 60–62). The taxonomic status of the remaining five cytotypes (A. cf. somalicus, 2n = 62 NFa = 62–63; ANI‐5, 2n = 56, NFa = 62; ANI‐6/6a 2n = 60, NFa = 72/76; ANI‐7, 2n = 56, NFa = 78; and ANI‐8, 2n = 44, NF = 72) is discussed. Finally, we reconstruct the phylogenetic relationships among all the known karyotypes on the basis of banding data available for the genus in Africa and show the occurrence of two main clades, each characterized by different types of chromosomal rearrangements. The times of the cladogenetic events, inferred by a molecular clock, indicate that karyotype evolution has accomplished almost all the dichotomic events from the end of the Miocene to the present day. The discovery of a large chromosomal differentiation between populations showing low genetic distances and intrapopulation chromosomal polymorphism suggests that the process of chromosomal differentiation in Arvicanthis is still ongoing and may possibly be responsible for speciation.     

Comparative cytogenetics of six Indo‐Pacific moray eels (Anguilliformes: Muraenidae) by chromosomal banding and fluorescence in situ hybridization

A comparative cytogenetic analysis, using both conventional staining techniques and fluorescence in situ hybridization, of six Indo‐Pacific moray eels from three different genera (Gymnothorax fimbriatus, Gymnothorax flavimarginatus, Gymnothorax javanicus, Gymnothorax undulatus, Echidna nebulosa and Gymnomuraena zebra), was carried out to investigate the chromosomal differentiation in the family Muraenidae. Four species displayed a diploid chromosome number 2n = 42, which is common among the Muraenidae. Two other species, G. javanicus and G. flavimarginatus, were characterized by different chromosome numbers (2n = 40 and 2n = 36). For most species, a large amount of constitutive heterochromatin was detected in the chromosomes, with species‐specific C‐banding patterns that enabled pairing of the homologous chromosomes. In all species, the major ribosomal genes were localized in the guanine‐cytosine‐rich region of one chromosome pair, but in different chromosomal locations. The (TTAGGG)_n telomeric sequences were mapped onto chromosomal ends in all muraenid species studied. The comparison of the results derived from this study with those available in the literature confirms a substantial conservation of the diploid chromosome number in the Muraenidae and supports the hypothesis that rearrangements have occurred that have diversified their karyotypes. Furthermore, the finding of two species with different diploid chromosome numbers suggests that additional chromosomal rearrangements, such as Robertsonian fusions, have occurred in the karyotype evolution of the Muraenidae.     

On the lectotype designation of Bryconamericus rubropictus (Berg) (Characiformes,Characidae, Stevardiinae)

The lectotype specimen of Bryconamericus rubropictus (Berg) and its designation have remained imprecisely documented since its publication. The lack of a photograph or an unambiguous illustration, correct size, inaccurate labelling, and proper specimen separation has led to an uncertainty about the identity and nomenclatural status of the lectotype. We recovered and provided detailed morphological data on the specimen that must be recognised as the lectotype. This contribution brings stability and clarity on the nomenclatural status of the species and its type material.     

Note on the karyotype and NOR phenotype of leuciscine fish Acanthobrama marmid (Osteichthyes, Cyprinidae)

The karyotype and major ribosomal sites as revealed using silver staining of Anatolian leuciscine cyprinid fish Acanthobrama marmid were studied. The diploid chromosome number was invariably 2n = 50. Karyotype consisted of eight pairs of metacentric, 13 pairs of submetacentric and four pairs of subtelocentric to acrocentric chromosomes. The largest chromosome pair of the complement was subtelo-to acrocentric characteristically, which is a characteristic cytotaxonomic marker for representatives of the cyprinid lineage Leuciscinae. The nucleolar organizer regions (NORs) were detected in the telomeres of two pairs of medium sized submeta-to subtelocentric chromosomes. No heteromorphic sex chromosomes were found. The karyotype pattern of A. marmid is nearly identical to that found in most other representatives of the Eurasian leuciscine cyprinids, while the multiple NOR phenotype appears to be more derived as opposed to a uniform one, ubiquitous in this group.     

Molecular characterization of different preproGnRHs in Astyanax altiparanae (Characiformes): Effects of GnRH on female reproduction

Gonadotropin‐releasing hormone (GnRH) is a key molecule in the initiation of the hypothalamic–pituitary–gonadal axis. Thus, knowledge about GnRH may contribute to the effectiveness of species reproduction. Using a Neotropical tetra Astyanax altiparanae as a fish model species, the GnRH forms were characterized at the molecular level and the role of injected GnRHs in vivo was evaluated. The full‐length complementary DNA (cDNA) sequences of preproGnRH2 (612 bp) and preproGnRH3 (407 bp) of A. altiparanae were obtained, and the GnRH1 form was not detected. The cDNA sequences of preproGnRH2 and preproGnRH3 were found to be conserved, but a change in the amino acid at position 8 of the GnRH3 decapeptide of A. altiparanae was observed. All the injected GnRHs stimulated lhβ messenger RNA (mRNA) expression but not fshβ mRNA expression, and only GnRH2 was able to increase maturation‐inducing steroid (MIS) levels and possibly stimulate oocyte release. Furthermore, only GnRH2 was able to start the entire reproductive hormonal cascade and induce spawning.     

An unusual ZZ/ZW sex chromosome system in Characidium fishes (Crenuchidae, Characiformes) with the presence of rDNA sites

Characidium fishes with a sex chromosome system form a monophyletic group. This work presents data of Characidium lanei from the South Atlantic basin (Brazil), including an unknown type of ZW sex chromosome system for the groups including the presence of rDNA sites on sex chromosomes.     

Recent chromosome diversification in the evolutionary radiation of the freshwater fish family Curimatidae (Characiformes)

Neotropical Curimatidae fishes include 97 species in eight genera. Basic cytogenetic studies show a karyotype of 2 n = 54 chromosomes in most species. Karyotype divergence of the nucleolus organizing regions between species has been reported, and these regions appear to be good cytotaxonomic markers. In the present work, karyotype, heterochromatin and Ag-NOR variability in 13 species were investigated to analyse the chromosome diversification in view of the biogeographic history of this group. Only Cyphocharax platanus showed a karyotype with 2 n = 58 chromosomes. Ag-NOR and C-banding patterns were quite divergent among the species studied. All species whose C-bands were analysed had heterochromatic blocks associated with the nucleolus organizing regions. Species with multiple Ag-NORs also showed an increase in NOR-associated heterochromatic blocks. C-banding showed considerable differentiation among species, revealing a pronounced chromosome diversification in this group. Karyotypic variability corroborates the hypothesis that these fishes in Amazon region show various discrete patterns of species endemism. Chromosome diversification in curimatids has a recent origin and appears to be accompanying the post-Andean speciation responsible for the diversity of species in the family.