Josiane Baccarin Traldi

Origin of the X1X1X2X2/X1X2Y sex chromosome system of Harttia punctata (Siluriformes, Loricariidae) inferred from chromosome painting and FISH with ribosomal DNA markers

AbstractHarttia is a genus in the subfamily Loricariinae that accommodates fishes popularly known as armored catfishes. They show extensive karyotypic diversity regarding interspecific numerical/structural variation of the karyotypes, with the presence of the XX/XY1Y2 multiple sex chromosome system, as found in H. carvalhoi. In this context, this study aimed to characterize Harttia punctata chromosomally, for the first time, and to infer the rearrangements that originated the X1X1X2X2/X1X2Y multiple sex chromosome system present in this species. The data obtained in this study, with classical (Giemsa, C-banding and AgNORs) and molecular methodologies (fluorescence in situ hybridization) and chromosome microdissection, indicated that a translocation between distinct acrocentric chromosomes bearing rRNA genes, accompanied by deletions in both chromosomes, might have originated the neo-Y chromosome in this species. The data also suggest that the multiple sex chromosome systems present in H. carvalhoi and H. punctata had an independent origin, evidencing the recurrence of chromosome alterations in species from this genus.

Comparative cytogenetics of three populations from the Rhamdia quelen species complex (Siluriformes, Heptapteridae) in two Brazilian hydrographic basins

Abstract Among the more than 30 families of the order Siluriformes, Heptapteridae is composed of 189 species distributed into 24 genera. Rhamdia, which has wide distribution throughout the Neotropical region, presents only 11 valid species, with 8 being in the Brazilian territory. Rhamdia quelen is the only species considered as widely distributed in almost all Brazilian hydrographic basins. It is a Neotropical fish species with a confusing taxonomical history. Classic and molecular cytogenetics data for three populations of this species from two large Brazilian hydrographic basins (Paraná and Araguaia) are presented here. The diploid number found for the three analyzed populations was 58 chromosomes, but with distinct karyotypic formulae. The presence of B chromosomes was detected in the two Araguaia River populations with intra- and interindividual variation. C-banding evidenced little heterochromatin in the three analyzed populations. FISH with 18S rDNA probes evidenced a single chromosome pair bearing this site, confirming the presence of simple NORs, as visualized through silver nitrate staining. The site of 5S rDNA was observed in only one pair of chromosomes, but differing in the marked pair and their location. Based in the differences of the karyotypic formulae and rDNA 5S found between populations on this study and many others available in the literature, it is suggested that this group represents a species complex, and that a new and detailed taxonomical review is necessary.

In situ Localization of (GATA)n and (TTAGGG)n Repeated DNAs and W Sex Chromosome Differentiation in Parodontidae (Actinopterygii: Characiformes)

The family Parodontidae presents a conserved diploid number of 54 chromosomes and different stages associated with ZW sex chromosome differentiation. For the great majority of species in this family it was proposed that the karyotypic diversification is mostly due to repetitive DNA mobility and accumulation. In this study, 2 repetitive probes, (GATA)n and (TTAGGG)n, were used to assess probable mechanisms of chromosome diversification, especially those related to molecular differentiation of the W chromosome. Results showed that the (GATA)n sequence is involved in the differentiation of the W chromosome female-specific region of Parodontidae and that it is accumulated in diverse autosomes. The (TTAGGG)n repeat is part of the vertebrate telomere, and the presence of interstitial telomeric sites may help to identify chromosome rearrangements. However, in Parodontidae, no interstitial telomeric sites were detected. This study shows plasticity in the amount of the (GATA)n repeat in Parodontidae that may be involved in chromatin modifications and transcriptional control of the W chromosome, and the role of repetitive DNAs in genomic diversification in this fish family is discussed.

The role of the Robertsonian rearrangements in the origin of the XX/XY1Y2 sex chromosome system and in the chromosomal differentiation in Harttia species (Siluriformes, Loricariidae)

Harttia is a genus of the subfamily Loricariinae that posses a broad chromosomal variation. In addition to interspecific karyotype diversity within this group, a multiple sex chromosome system, XX/XY1Y2, has been described for Harttia carvalhoi. Thus, this study aimed to determine the role of chromosomal rearrangements in karyotype differentiation in Harttia by classical and molecular cytogenetic procedures. The results show that Robertsonian rearrangements have a prominent role in the chromosomal diversification of the species analysed, which initially leads to hypothesize a diploid number reduction in Harttia torrenticola and H. carvalhoi. The metacentric chromosome 1, shared between H. torrenticola and H. carvalhoi, could have originated from centric fusions from the ancestral karyotype. A centric fission event associated with the first metacentric pair allowed for the origination of a multiple sex chromosome system XX/XY1Y2, specific to H. carvalhoi. This study highlights the relevance of Robertsonian rearrangements in karyotypic differentiation of the species studied and demonstrates that the occurrence of a centric fission, as opposed to a previously hypothesised chromosome fusion, is directly implicated in the origin of the sex chromosome system of H. carvalhoi.

Physical mapping of (GATA)n and (TTAGGG)n sequences in species of Hypostomus (Siluriformes, Loricariidae).

Hypostomus, a genus of Loricariidae, is one of the most diversified and widely distributed genera among fishes of South America (Weber 2003). It is the dominant genus of armoured catfish in Brazilian rivers (Weber 2003). Chromosomally, this group is highly diversified and studies of repetitive DNAs in these species are still in preliminary stages. Hypostomus karyotypes range from 54 chromosomes in H. plecostomus (Muramoto et al. 1968) to 84 chromosomes in Hypostomus sp. 2 (Cereali et al. 2008). Artoni and Bertollo (2001) consider the diploid number of 54 chromosomes as basal for Hypostominae, using Trichomycteridae as outgroup, suggesting that the chromosome evolution of Hypostomus occurred through centric fissions. Recently, Bueno et al. (2012) showed that other chromosomal rearrangements, such as inversions, deletions, duplications and heterochromatinization, could contribute to the chromosomal differentiation within the genus. Repetitive DNAs arranged in tandem are important tools in studies of taxonomic and evolutionary problems. These sequences could be associated with chromosomal rearrangement events (Wichman et al. 1991; Rosa et al. 2012; and others). According to Wichman et al. (1991), satellite DNAs rapidly diverge during evolution. Thereby, they are important for solving the taxonomic and evolutionary problems among related species.

Chromosomal characterization of armored catfish Harttia longipinna (Siluriformes, Loricariidae): first report of B chromosomes in the genus.

The B chromosomes are accessory elements that are widely distributed among eukaryotic genomes and often show non-Mendelian inheritance. They are considered dispensable for the growth, development, and reproduction of organisms. Some studies have suggested that these elements may affect sex determination. Harttia is a small armored catfish genus that shows sexual dimorphism, including hypertrophied odontodes on the pectoral fin spines and along the margins of the snout in mature males. They exhibit considerable karyotypic diversity with diploid number (2n) variation and heteromorphic sex system in H. carvalhoi. To date, no occurrences of B chromosomes in the Harttia genus were detected and no relation to sexual differentiation in Neotropical fish has been determined. To determine the validity of this claim, the present paper characterized specimens of Harttia longipinna by classical and molecular cytogenetic methods. The 2n found was 58 (16m + 12sm + 16st + 14a), but of the 50 specimens analyzed (30 male and 20 female), 23 specimens (16 males and seven females) show an intra-individual from 0 to 2 micro B chromosomes. The B chromosomes were completely heterochromatic. The single NORs were shown in the first acrocentric pair with silver staining and 18S rDNA probing. FISH performed with 5S rDNA probe showed a single cistron in the proximal region of the short arm of a small metacentric pair. Thus, the cytogenetic data obtained in this study of H. longipinna highlight the karyotypic diversity found within the genus Harttia, and represent the first description of B chromosomes for this genus.

Comparative Cytogenetics of Hoplerythrinus unitaeniatus (Agassiz, 1829) (Characiformes, Erythrinidae) Species Complex from Different Brazilian Hydrographic Basins

Chromosomal characteristics of Hoplerythrinus unitaeniatus populations from 5 Brazilian river basins, namely Arinos (Amazonas basin), Araguaia, Paraguai, Alto Paraná, and São Francisco were analyzed by conventional Giemsa staining, C-banding, silver nitrate impregnation, and fluorescence in situ hybridization (FISH) with 18S and 5S rDNA and telomeric sequence (TTAGGG)n probes. The diploid chromosome number was 2n = 48 in representatives of the populations from Paraguai and Alto Paraná River basins and 2n = 52 for those from the Arinos and Araguaia River basins. The São Francisco population had individuals with 2n = 50 and 52 occurring in sympatry. C-banding showed heterochromatic blocks mainly located at interstitial and pericentromeric positions in most of the chromosomes. Silver nitrate impregnation demonstrated simple NORs for representatives from Arinos and Araguaia River populations and multiple NORs for specimens from Paraguai, Alto Paraná, and São Francisco River populations. FISH with 18S and 5S rDNA probes revealed many chromosomes carrying these cistrons, with up to 21 chromosomes bearing 18S rDNA sites (Alto Rio Paraná basin) and up to 12 chromosomes with 5S rDNA sites (Paraguai basin), besides the occurrence of colocalization in all populations. FISH with telomeric sequence (TTAGGG)n detected sites in the terminal portion of the chromosomes in all populations. These data reinforce the idea that H. unitaeniatus is a species complex. Evolutionary and biogeographical aspects of the group in the Neotropical region are discussed.

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin

The Iguazu river is a tributary of the left margin of the Paraná river, isolated from this basin about 22 million years ago with the appearance of the Iguazu Falls. The Iguazu river is characterized by high endemism due to two factors: its rugged topography and the old isolation caused by formation of the Iguazu Falls. This study analyzed cytogenetically a population of Glanidium ribeiroi collected in a region at the final stretch of this basin, by Giemsa staining, C-banding, impregnation by silver nitrate, and FISH with probes of 5S rDNA, 18S rDNA, telomeric sequence [TTAGGG]n, and [GATA]n repeats. The diploid number was equal to 58 chromosomes. The heterochromatin was present in the terminal region of almost all chromosomes. The Ag-NORs were simple and presented interstitially on the short arm of the submetacentric pair 14, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA-FISH marked only the submetacentric pair 16 on the long arm in interstitial position. The FISH with [TTAGGG]n probe presented all telomeres labeled as expected, with an absence of Interstitial Telomeric Sequence (ITS). The repetitive [GATA]n sequence was dispersed throughout the genome, with preferential location in the terminal region of all chromosomes. The data obtained are discussed herein with other species of Auchenipteridae, and other previously analyzed populations of G. ribeiroi from the Iguazu river, verifying differences among these populations, which should be mainly related to the rugged topography of this basin.

Chromosomal Mapping of Repeat DNA in Bergiaria westermanni (Pimelodidae, Siluriformes): Localization of 45S rDNA in B Chromosomes

The occurrence of repetitive DNA in autosomes and B chromosomes of Bergiaria westermanni was examined using conventional and molecular cytogenetic techniques. This species exhibited 2n = 56 chromosomes, with intra- and interindividual variation in the number of heterochromatic B chromosomes (from 0 to 4). The 5S rDNA was localized in pairs 1 and 5, and histone probes (H1, H3, and H4) and U2 small nuclear RNA were syntenic with 5S rDNA in pair 5. Histone sequences were also located in chromosome pair 14. The (GATA)n sequence was dispersed throughout the autosomes and B chromosomes, with clusters (microsatellite accumulation) in some chromosome regions. The telomeric probe revealed no signs of chromosomal rearrangements in the genome of B. westermanni. The 45S rDNA sites were detected in the terminal region of pair 27; these sites corresponded to a GC-rich heterochromatin block. In addition, 3 of the 4 B chromosomes also contained 45S rDNA copies. Silver nitrate staining in interphase nuclei provided indirect evidence of the expression of these rRNA genes in B chromosomes, indicating the probable origin of these elements. This report shows plasticity in the chromosomal localization of repeat DNA in B. westermanni and features a discussion of genomic diversification.