Restriction endonuclease banding and digestion resistant heterochromatin in triatomines, genus Panstrongylus

The present work realized a comparative study in meiosis of two triatomines, Panstrongylus herreri and P. megistus, by cytogenetic techniques involving the restriction endonucleases Hae III and Alu I and C-banding. The system of sex chromosomes in Panstrongylus is of the X1X2Y type, and experiments corroborated the common origin hypothesis of the X chromosomes by fragmentation of single X. In both species the restriction endonucleases (RE) presented banding patterns in part similar to C-banding. However, in some early meiotic phases it was possible to verify differentiation of the heterochromatic pattern. This work suggests that other elements besides presence of recognition sites, such as chromatin packing degree and DNA-protein interaction, act in RE results, since digestion patterns occur in early spermatogenesis. However, metaphase chromosomes were practically inaccessible to the endonucleases.

Cytogenetics of two species of Paratelmatobius (Anura: Leptodactylidae), with phylogenetic comments

In this paper we provide a cytogenetic analysis of Paratelmatobius cardosoi and Paratelmatobius poecilogaster. The karyotypes of both species showed a diploid number of 24 chromosomes and shared some similarity in the morphology of some pairs. On the other hand, pairs 4 and 6 widely differed between these complements. These karyotypes also differed in their NOR number and location. Size heteromorphism was seen in all NOR-bearing chromosomes of the two karyotypes. In addition, both karyotypes showed small centromeric C-bands and a conspicuous heterochromatic band in the short arm of chromosome 1, although with a different size in each species. The P. cardosoi complement also showed other strongly stained non-centromeric C-bands, with no counterparts in the P. cardosoi karyotype. Chromosome staining with fluorochromes revealed heterogeneity in the base composition of two of the non-centromeric C-bands of P. cardosoi. Comparison of the chromosomal morphology of these Paratelmatobius karyotypes with that of P. lutzii showed that the P. poecilogaster karyotype is more similar to that of P. lutzii than P. cardosoi. These cytogenetic results agree with the proposed species arrangements in the P. cardosoi and P. lutzii groups based on morphological and ecological data.

Cytogenetic analysis of five species of the subfamily Tetragonopterinae (Teleostei, Characiformes, Characidae)

The subfamily Tetragonopterinae is composed by a large number of species distributed in South and Central America. This subfamily has many taxonomic and phylogenetic problems, being considered by several authors as an artificial group. With the objective to better understanding the relationships among the components of this fish group, cytogenetic studies were conduced on five species of Tetragonopterinae. Astyanax janeiroensis had 2n=50 chromosomes (6M+14SM+14ST+16A), Hyphessobrycon reticulatus had 2n=50 chromosomes (14M+20SM+16ST), Hollandichthys multifasciatus had 2n=50 chromosomes (10M+12SM+28ST), Ctenobrycon hauxwellianus had 2n=50 chromosomes (10M+6SM+34ST), and Phenacogaster cf. pectinatus had 2n=46 chromosomes (12M+2ST+32A). Only A. janeiroensis had multiple NORs, while all other species had simple NORs. Small heterochromatic blocks were observed in the chromosomes of all species in a pericentromeric position. A. janeiroensis also had some chromosomes with large heterochromatic blocks in a terminal position and a pair with an interstitial block. The karyotypic evolution of each genus is discussed.

High content of constitutive heterochromatin in two species of Pseudonannolene (Diplopoda)

Due to the fact that a considerable amount of constitutive heterochromatin has been commonly found in millipedes, this paper presents the characterization of the chromosomal complement, through C-banding technique, of two species of millipedes belonging to the genus Pseudonannolene. The species Pseudonannolene tocaiensis shows 2n=20, with five chromosomal pairs almost completely heterochromatic and the remaining pairs with pericentromeric C+ blocks. P. silvestris shows 2n=16, with a large heterochromatic portion in the first two pairs of autosomes, which presented the long arms totally C+.

High content of constitutive heterochromatin in two species of Pseudonannolene (Diplopoda)

Due to the fact that a considerable amount of constitutive heterochromatin has been commonly found in millipedes, this paper presents the characterization of the chromosomal complement, through C-banding technique, of two species of millipedes belonging to the genus Pseudonannolene. The species Pseudonannolene tocaiensis shows 2n=20, with five chromosomal pairs almost completely heterochromatic and the remaining pairs with pericentromeric C+ blocks. P. silvestris shows 2n=16, with a large heterochromatic portion in the first two pairs of autosomes, which presented the long arms totally C+.

Cytogenetic analysis of three sympatric Gymnotus species (Teleostei: Gymnotidae) from the Fundo Stream, MG, Brazil

Three sympatric species of Gymnotus from the Fundo stream, a small tributary of the Sapucai river, Minas Gerais State, Brazil, were studied in relation to their karyology. Gymnotus sylvius presented 2n=40 chromosomes (36 m/sm+4 st/a), Gymnotus sp. presented 2n=50 (26 m/sm+ 24 st/a) and Gymnotus paraguensis had 2n=54 (50 m/sm+4 st/a). C-banding demonstrated positively stained heterochromatic blocks in the centromeric position of few chromosomes on G. sylvius and in the centromeric region of all chromosomes on G. paraguensis samples. The nucleolus organizer region (NOR) was located on the short arm of one st chromosome pair in G. sylvius and Gymnotus sp., and in the interstitial position on the short arm of the pair number one and below the centromere on a third chromosome on G. paraguensis. The cytogenetic data obtained indicate that Gymnotus sp. represent a new Gymnotus specie with a karyotypic constitution never observed on others species from this genus. Some aspects related to the chromosome diversification of these Gymnotus are discussed. © 2007 The Japan Mendel Society.

Chromosome mapping of repetitive sequences in Anostomidae species: Implications for genomic and sex chromosome evolution

Background: Members of the Anostomidae family provide an interesting model system for the study of the influence of repetitive elements on genome composition, mainly because they possess numerous heterochromatic segments and a peculiar system of female heterogamety that is restricted to a few species of the Leporinus genus. The aim of this study was to isolate and identify important new repetitive DNA elements in Anostomidae through restriction enzyme digestion, followed by cloning, characterisation and chromosome mapping of this fragment. To identify repetitive elements in other Leporinus species and expand on studies of repetitive elements in Anostomidae, hybridisation experiments were also performed using previously described probes of LeSpeI repetitive elements. Results: The 628-base pair (bp) LeSpeII fragment was hybridised to metaphase cells of L. elongatus individuals as well as those of L. macrocephalus, L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii and S. isognathus. In L. elongatus, both male and female cells contained small clusters of LeSpeII repetitive elements dispersed on all of the chromosomes, with enrichment near most of the terminal portions of the chromosomes. In the female sex chromosomes of L. elongatus (Z2,Z2/W1W 2), however, this repeated element was absent. In the remaining species, a dispersed pattern of hybridisation was observed on all chromosomes irrespective of whether or not they were sex chromosomes. The repetitive element LeSpeI produced positive hybridisations signals only in L. elongatus, L. macrocephalus and L. obtusidens, i.e., species with differentiated sex chromosomes. In the remaining species, the LeSpeI element did not produce hybridisation signals. Conclusions: Results are discussed in terms of the effects of repetitive sequences on the differentiation of the Anostomidae genome, especially with respect to sex chromosome evolution. LeSpeII showed hybridisation patterns typical of Long Interspersed Elements (LINEs). The differential distribution of this element may be linked to sex chromosome differentiation in L. elongatus species. The relationship between sex chromosome specificity and the LeSpeI element is confirmed in the species L. elongatus, L. macrocephalus and L. obtusidens. © 2012 da Silva et al.; licensee BioMed Central Ltd.

Cytogenetic analysis in Thoracocharax stellatus (Kner, 1858) (Characiformes, Gasteropelecidae) from Paraguay river basin, Mato Grosso, Brazil

Thoracocharax stellatus (Characiformes, Gasteropelecidae) is a small Neotropical species of fish, widely distributed in several rivers of South America. Evidence for karyotype heteromorphysm in populations from different geographical regions has been reported for this species. In this way, populations of T. stellatus from the Paraguay River basin were cytogenetically characterized and the results were compared with other studies performed in the same species but from different basins. The results showed a diploid number of 2n = 54 for T. stellatus, with chromosomes arranged in 6 metacentric (m), 6 submetacentric (sm), 2 subtelocentric (st) and 40 acrocentric (a), for both sexes, with a simple Nucleolus Organiser Region (NOR) system reported by the techniques of silver nitrate impregnation and fluorescent in situ hybridisation (FISH) using 18S rDNA sequences as probe. The distribution of constitutive heterochromatin, observed by the C-band technique and Chromomycin A3 staining showed great similarity among the analyzed populations and consists mainly of discrete blocks in the pericentromeric and telomeric regions of most chromosomes. The presence of female heterogamety was alsoobserved indicating a ZZ/ZW system with W chromosome almost totally heterochromatic. The results also show cytogenetic diversity of the group and are useful to understand the mechanisms of karyotype evolution of the family. © Edson Lourenço da Silva et al.

Chromosomal evolution of neotropical cichlids: The role of repetitive DNA sequences in the organization and structure of karyotype

Cichlids are important in the aquaculture and ornamental fish trade and are considered models for evolutionary biology. However, most studies of cichlids have investigated African species, and the South American cichlids remain poorly characterized. Studies in neotropical regions have focused almost exclusively on classical cytogenetic approaches without investigating physical chromosomal mapping of specific sequences. The aim of the present study is to investigate the genomic organization of species belonging to different tribes of the subfamily Cichlinae (Cichla monoculus, Astronotus ocellatus, Geophagus proximus, Acaronia nassa, Bujurquina peregrinabunda, Hoplarchus psittacus, Hypselecara coryphaenoides, Hypselecara temporalis, Caquetaia spectabilis, Uaru amphiacanthoides, Pterophyllum leopoldi, Pterophyllum scalare, and Symphysodon discus) and reexamine the karyotypic evolutionary patterns proposed for this group. Variations in some cytogenetic markers were observed, although no trends were found in terms of the increase, decrease, or maintenance of the basal diploid chromosome number 2n = 48 in the tribes. Several species were observed to have 18S rDNA genetic duplications, as well as multiple rDNA loci. In most of the taxa analyzed, the 5S rDNA was located in the interstitial region of a pair of homologous chromosomes, although variations from this pattern were observed. Interstitial telomere sites were also observed and appear to be involved in chromosomal rearrangement events and the accumulation of repeat-rich satellite DNA sequences. Our data demonstrated the karyotypic diversity that exists among neotropical cichlids, suggesting that most of this diversity is due to the repetitive sequences present in heterochromatic regions and that repeat sequences have greatly influenced the karyotypic evolution of these fishes. © 2012 Springer Science+Business Media B.V.

Distribution of constitutive heterochromatin in two species of triatomines: Triatoma lenti Sherlock and Serafim (1967) and Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata (2002)

Triatoma lenti and Triatoma sherlocki are hemipterans that belong to the brasiliensis subcomplex. In triatomines, the constitutive heterochromatin pattern is species-specific and allows, in many cases, for the grouping of species. Thus, we cytogenetically analyzed T. sherlocki and T. lenti using C-banding, and we compared the results with previous ones obtained in other species of the brasiliensis subcomplex. Both species were found to have a male diploid chromosome number of 22 chromosomes (2n = 20A. +. XY) with heterochromatic blocks at one or both chromosomal ends of all autosomal pairs. During early meiotic prophase, they showed a large heteropycnotic chromocenter constituted by the association of both sex chromosomes plus two autosomal pairs and many heterochromatic blocks dispersed inside the nucleus. All of these cytogenetic characteristics are similar to those observed in other species of brasiliensis subcomplex, results which confirm the grouping of T. sherlocki and T. lenti within this subcomplex. However, we emphasize the importance of other approaches, such as molecular analysis, to confirm the placement of T. lenti within the brasiliensis subcomplex. © 2012 Elsevier B.V.

Chromosome painting of Z and W sex chromosomes in Characidium (Characiformes, Crenuchidae)

Some species of the genus Characidium have heteromorphic ZZ/ZW sex chromosomes with a totally heterochromatic W chromosome. Methods for chromosome microdissection associated with chromosome painting have become important tools for cytogenetic studies in Neotropical fish. In Characidium cf. fasciatum, the Z chromosome contains a pericentromeric heterochromatin block, whereas the W chromosome is completely heterochromatic. Therefore, a probe was produced from the W chromosome through microdissection and degenerate oligonucleotide-primed polymerase chain reaction amplification. FISH was performed using the W probe on the chromosomes of specimens of this species. This revealed expressive marks in the pericentromeric region of the Z chromosome as well as a completely painted W chromosome. When applying the same probe on chromosome preparations of C. cf. gomesi and Characidium sp., a pattern similar to C. cf. fasciatum was found, while C. cf. zebra, C. cf. lagosantense and Crenuchus spilurus species showed no hybridization signals. Structural changes in the chromosomes of an ancestral sexual system in the group that includes the species C. cf. gomesi, C. cf. fasciatum and Characidium sp., could have contributed to the process of speciation and could represent a causal mechanism of chromosomal diversification in this group. The heterochromatinization process possibly began in homomorphic and homologous chromosomes of an ancestral form, and this process could have given rise to the current patterns found in the species with sex chromosome heteromorphism. © 2013 Springer Science+Business Media Dordrecht.

Evolutionary dynamics of retrotransposable elements Rex1, Rex3 and Rex6 in neotropical cichlid genomes

Background: Transposable elements (TEs) have the potential to produce broad changes in the genomes of their hosts, acting as a type of evolutionary toolbox and generating a collection of new regulatory and coding sequences. Several TE classes have been studied in Neotropical cichlids; however, the information gained from these studies is restricted to the physical chromosome mapping, whereas the genetic diversity of the TEs remains unknown. Therefore, the genomic organization of the non-LTR retrotransposons Rex1, Rex3, and Rex6 in five Amazonian cichlid species was evaluated using physical chromosome mapping and DNA sequencing to provide information about the role of TEs in the evolution of cichlid genomes. Results: Physical mapping revealed abundant TE clusters dispersed throughout the chromosomes. Furthermore, several species showed conspicuous clusters accumulation in the centromeric and terminal portions of the chromosomes. These TE chromosomal sites are associated with both heterochromatic and euchromatic regions. A higher number of Rex1 clusters were observed among the derived species. The Rex1 and Rex3 nucleotide sequences were more conserved in the basal species than in the derived species; however, this pattern was not observed in Rex6. In addition, it was possible to observe conserved blocks corresponding to the reverse transcriptase fragment of the Rex1 and Rex3 clones and to the endonuclease of Rex6. Conclusion: Our data showed no congruence between the Bayesian trees generated for Rex1, Rex3 and Rex6 of cichlid species and phylogenetic hypothesis described for the group. Rex1 and Rex3 nucleotide sequences were more conserved in the basal species whereas Rex6 exhibited high substitution rates in both basal and derived species. The distribution of Rex elements in cichlid genomes suggests that such elements are under the action of evolutionary mechanisms that lead to their accumulation in particular chromosome regions, mostly in heterochromatins. © 2013 Schneider et al.; licensee BioMed Central Ltd.

Tracking the evolution of sex chromosome systems in Melanoplinae grasshoppers through chromosomal mapping of repetitive DNA sequences

Background: The accumulation of repetitive DNA during sex chromosome differentiation is a common feature of many eukaryotes and becomes more evident after recombination has been restricted or abolished. The accumulated repetitive sequences include multigene families, microsatellites, satellite DNAs and mobile elements, all of which are important for the structural remodeling of heterochromatin. In grasshoppers, derived sex chromosome systems, such as neo-XY♂/XX♀ and neo-X1X2Y♂/X 1X1X2X2♀, are frequently observed in the Melanoplinae subfamily. However, no studies concerning the evolution of sex chromosomes in Melanoplinae have addressed the role of the repetitive DNA sequences. To further investigate the evolution of sex chromosomes in grasshoppers, we used classical cytogenetic and FISH analyses to examine the repetitive DNA sequences in six phylogenetically related Melanoplinae species with X0♂/XX♀, neo-XY♂/XX♀ and neo-X1X2Y♂/X1X1X 2X2♀ sex chromosome systems. Results: Our data indicate a non-spreading of heterochromatic blocks and pool of repetitive DNAs (C 0 t-1 DNA) in the sex chromosomes; however, the spreading of multigene families among the neo-sex chromosomes of Eurotettix and Dichromatos was remarkable, particularly for 5S rDNA. In autosomes, FISH mapping of multigene families revealed distinct patterns of chromosomal organization at the intra- and intergenomic levels. Conclusions: These results suggest a common origin and subsequent differential accumulation of repetitive DNAs in the sex chromosomes of Dichromatos and an independent origin of the sex chromosomes of the neo-XY and neo-X1X2Y systems. Our data indicate a possible role for repetitive DNAs in the diversification of sex chromosome systems in grasshoppers. © 2013Palacios-Gimenez et al.; licensee BioMed Central Ltd.

Estrutura e evolução cariotípica de peixes ciclídeos sul americanos

Pós-graduação em Biologia Geral e Aplicada - IBB

Comparação cariotípica entre Mazama gouazoubira e Mazama nemorivaga (Artiodactyla; Cervidae) por meio de marcadores citogenéticos clássicos, fish telomérica e pintura cromossômica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)