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.

Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) along the Iguazu river basin / Variação cariotípica de Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae) ao longo da bacia do rio Iguaçu

Abstract 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.

Resumo O rio Iguaçu é um afluente da margem esquerda do rio Paraná, que foi separado desta bacia a aproximadamente 22 milhões de anos com o surgimento das Cataratas do Iguaçu. Esse rio é caracterizado por elevado endemismo, o que se deve a dois fatores: sua acidentada topografia e ao antigo isolamento proporcionado pela formação das cataratas. No presente trabalho foi analisado cromossomicamente uma população de Glanidium ribeiroi coletada em uma região que corresponde ao trecho final desse rio, através de coloração com Giemsa, bandamento-C, impregnação pelo nitrato de prata e FISH com sondas de rDNA 5S, rDNA 18S, sequência telomérica [TTAGGG]n e repetições [GATA]n. O número diploide encontrado foi igual a 58 cromossomos. A heterocromatina se mostrou dispersa na região terminal de quase todos os cromossomos. As Ag-RONs são simples e presentes no braço curto em posição intersticial do par submetacêntrico 14, o que foi confirmado pela FISH com rDNA 18S. O rDNA 5S marcou apenas o par submetacêntrico 16 no braço longo em posição intersticial. A hibridização com sonda [TTAGGG]n revelou todos os telômeros marcados conforme esperado e ausência de Sequência Telomérica Intersticial (ITS). As repetições [GATA]n se apresentaram dispersas no genoma da espécie, com preferencial localização na região terminal de todos os cromossomos. Os dados aqui obtidos são discutidos com os de outras espécies de Auchenipteridae, especialmente de G. ribeiroi anteriormente analisados do rio Iguaçu. Diferenças populacionais são constatadas em decorrência do isolamento geográfico ocasionado pelas inúmeras cachoeiras existentes no curso do rio Iguaçu.

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

Abstract 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.

Resumo O rio Iguaçu é um afluente da margem esquerda do rio Paraná, que foi separado desta bacia a aproximadamente 22 milhões de anos com o surgimento das Cataratas do Iguaçu. Esse rio é caracterizado por elevado endemismo, o que se deve a dois fatores: sua acidentada topografia e ao antigo isolamento proporcionado pela formação das cataratas. No presente trabalho foi analisado cromossomicamente uma população de Glanidium ribeiroi coletada em uma região que corresponde ao trecho final desse rio, através de coloração com Giemsa, bandamento-C, impregnação pelo nitrato de prata e FISH com sondas de rDNA 5S, rDNA 18S, sequência telomérica [TTAGGG]n e repetições [GATA]n. O número diploide encontrado foi igual a 58 cromossomos. A heterocromatina se mostrou dispersa na região terminal de quase todos os cromossomos. As Ag-RONs são simples e presentes no braço curto em posição intersticial do par submetacêntrico 14, o que foi confirmado pela FISH com rDNA 18S. O rDNA 5S marcou apenas o par submetacêntrico 16 no braço longo em posição intersticial. A hibridização com sonda [TTAGGG]n revelou todos os telômeros marcados conforme esperado e ausência de Sequência Telomérica Intersticial (ITS). As repetições [GATA]n se apresentaram dispersas no genoma da espécie, com preferencial localização na região terminal de todos os cromossomos. Os dados aqui obtidos são discutidos com os de outras espécies de Auchenipteridae, especialmente de G. ribeiroi anteriormente analisados do rio Iguaçu. Diferenças populacionais são constatadas em decorrência do isolamento geográfico ocasionado pelas inúmeras cachoeiras existentes no curso do rio Iguaçu.

Chromosomal diversity in Hypostomus (Siluriformes, Loricariidae) with emphasis on physical mapping of 18S and 5S rDNA sites.

We examined chromosomes of three species of the genus Hypostomus, in order to contribute to the understanding of the karyotype evolution of this group. Specimens of H. ancistroides and H. nigromaculatus displayed differences in karyotype formulas, distribution and location of heterochromatin and nucleolus organizer regions when compared to other populations of the same species. We made the first cytogenetic characterization of H. tapijara, an endemic species in the Ribeira de Iguape River. These specimens had 2n = 66 chromosomes, while H. ancistroides showed 2n = 68 and H. nigromaculatus 2n = 76 chromosomes. Physical mapping of 18S and 5S rDNA sites of the three species showed simple, multiple and syntenic clusters. Synteny of ribosomal sites was found in H. ancistroides and H. tapijara, and an interspersed pattern between these sites in all chromosomes bearing the synteny was observed. We conclude that the genus Hypostomus has a high chromosome complexity that is accompanied by great morphological variation. It is evident that this group comprises an interesting model for understanding the chromosome evolution of Neotropical ichthyofauna.

Propidium iodide for making heterochromatin more evident in the C-banding technique.

The detection of regions of heterochromatin has been the subject of intense investigation. We investigated an adaptation of the commonly used technique by replacing the nonfluorescent dye, Giemsa, by a fluorescent one, propidium iodide. This adaptation produces greater contrast of the heterochromatic bands in metaphase chromosomes and can be especially valuable when the organisms studied possess heterochromatin that is pale and difficult to visualize. We discuss the interactions of these two dyes with DNA and the excitation of the fluorescent dye when irradiated with ultraviolet light.

A new technique for obtaining mitotic chromosome spreads from fishes in the field.

This study presents an adaptation of current methodologies for preparing mitotic chromosomes from fishes, optimized for use in the field. The high-quality preparations obtained using this modified methodology is suitable for subsequent chromosomal analysis. Importantly, this method is particularly useful when specimen collection sites are far from research laboratories or when researchers are working with highly sensitive species that do not survive long outside of their natural habitats.

Comparative cytogenetics of giant trahiras Hoplias aimara and H. intermedius (Characiformes, Erythrinidae): chromosomal characteristics of minor and major ribosomal DNA and cross-species repetitive centromeric sequences mapping differ among morphologically identical karyotypes.

Karyotype and cytogenetic characteristics of 2 species of giant trahiras, Hopliasintermedius, São Francisco river basin, and Hopliasaimara, Arinos river (Amazon basin), were examined by conventional (C-banding, Ag-NOR, DAPI/CMA(3) double-staining) and fluorescent in situ hybridization (FISH) with 5S, 18S rDNA probes and cross-species Cot-1 DNA probing. Both species invariably had diploid chromosome number 2n = 50 and identical karyotypes composed of 10 pairs of metacentric and 15 pairs of submetacentric chromosomes. On the other hand, staining with base-specific fluorochromes (CMA(3), DAPI) and FISH mapping of repetitive DNA sequences showed extensive interspecific differences: while the genome of H. aimara had one submetacentric pair bearing CMA(3)-positive (DAPI-negative) sites, that of H. intermedius had 4 such pairs; while FISH with a 5S rDNA probe showed one (likely homologous) signal-bearing pair, that with 18S rDNA displayed one signal-bearing pair in H. intermedius and 2 such pairs in H. aimara. Cross-species FISH probing with Cot-1 DNA prepared from total DNA of both species showed no signals of Cot-1 DNA from H. aimara on chromosomes of H. intermedius but reciprocally (Cot-1 DNA from H. intermedius on chromosomes of H. aimara) displayed signals on at least 4 chromosome pairs. Present findings indicate (i) different composition of repetitive sequences around centromeres, (ii) different NOR phenotypes and (iii) distinct taxonomic status of both giant trahira species.