Cytogenetic characterization of F1, F2 and backcross hybrids of the Neotropical catfish species Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum (Pimelodidae, Siluriformes)

The cytogenetic characteristics of Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum and their F1, F2 and backcross hybrids were assessed by using chromosome banding techniques. The diploid number of 56 chromosomes was constant in all species and lineages, with a karyotypic formula containing 20 metacentric, 12 submetacentric, 12 subtelocentric and 12 acrocentric chromosomes. Nucleolar organizer regions (NORs) were identified in two subtelocentric chromosomes in the parents and hybrids, with partial nucleolar dominance in F1 and F2 specimens. Heterochromatic blocks were detected in the terminal and centromeric regions of some chromosomes in all individuals. For parental and hybrid lineages, 18S ribosomal clusters corresponding to NORs and 5S ribosomal genes were identified in distinct pairs of chromosomes. The striking conservation in the chromosomal macrostructure of the parental species may account for the fertility of their F1 hybrids. Similarly, the lack of marked alterations in the chromosomal structure of the F1 hybrids could account for the maintenance of these features in post-F1 lineages

Supernumerary chromosomes in the pufferfish Sphoeroides spengleri: first occurrence in marine Teleostean Tetraodontiformes fish

Cytogenetic analyses carried out in eight specimens of Sphoeroides spengleri revealed the presence of 2n = 46 chromosomes (20 M/SM and 26 ST/A). Besides the standard karyotypical set, the presence of B microchromosomes was observed in two individuals, ranging from 0 to 2 microchromosomes per cell. A karyotype composed by 2n = 46 chromosomes with occurrence of M and SM chromosomes is considered basal for the species from the clade comprising the families Tetraodontidae, Balistidae, and Diodontidae, although it represents a derived condition for the order Tetraodontiformes, whose basal karyotype would be composed by 2n = 48 acrocentric chromosomes. The occurrence of B microchromosomes in marine Tetraodontiformes fish was not known, and this represents the first report of such a chromosomal type.243245Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Cytogenetic Tools to Study the Biodiversity of Neotropical Fish: From the Classic to the Advent of Cell Culture

Neotropical Ichthyofauna is considered the richest and most diverse in the world. All this biodiversity has attracted attention from researchers from different areas of study, including the cytogenetics. Many cytogenetics studies have search to understand the evolution of macro and micro karyotype structure of these different groups of fish, and classical and molecular cytogenetics techniques have contributed significantly for all knowledge of this karyotypic diversity. Recently, the use of cell cultures as an alternative to obtaining mitotic chromosomes opening up new opportunities to study groups that have not been explored or have not yet been cytogenetically investigated. In this work, we take a chronological overview of the advances of different cytogenetic techniques (“in vivo” and “in vitro” methods to obtain the chromosome, C-banding, the detection of nucleolar organizer regions (Ag-RON), fluorescent in situ hybridization (FISH) with several repetitive probes and paint chromosome) over the decades and how these techniques helped elucidate questions of the organization and function of the fish genome

Karyotype diversity and patterns of chromosomal evolution in Eigenmannia (Teleostei, Gymnotiformes, Sternopygidae)

Conventional (Giemsa, C-banding, Ag – NORs) and molecular [5S rDNA, 18S rDNA, (TTAGGG)n] cytogenetic techniques were employed to study six species of the genus Eigenmannia Jordan & Evermann, 1896. They exhibited diploid chromosome numbers ranging from 2n=28 (Eigenmannia sp.1) to 2n=38 (E. virescens (Valenciennes, 1836)). The C-banding results revealed that species with the lowest 2n have less heterochromatin content and that morphologically differentiated sex chromosomes observed in two species showed distinct patterns of heterochromatin. While the X1, X2 and Y-chromosomes of Eigenmannia sp.2 showed only centromeric heterochromatin, the XY sex chromosomes of E. virescens possessed large heterochromatic blocks in the terminal position, particularly on the X chromosome. The nucleolus organizer regions (NORs) were located in different positions when compared to the 5S rDNA sites. Additionally, the presence of minor ribosomal gene sites on the sex chromosome pair of E. virescens represented a new type of the sex chromosomes in this group. The telomeric probe (TTAGGG)n hybridized to the terminal portion of all chromosomes in all species examined however, interstitial telomeric sites were found in the metacentric pair No. 2 in Eigenmannia sp.1. The analyzes confirmed some hypotheses about karyotype evolution in the genus Eigenmannia, and brought new information about the distribution of the genetic material in the chromosomes of the samples analyzed providing new insights for understanding the process differentiation in the genomes of species under study

Genetic variability of two populations of Pseudoplatystoma reticulatum from the Upper Paraguay River Basin

Catfishes of the genus Pseudoplatystoma are very important species due to both their high commercial value and their ecological role as voracious predators. They undertake lengthy migratory movements during their life-cycle, this including reproductive migration which occurs from October to December in the rainy season. In the present study, seven microsatellite loci were analyzed to access genetic variability in two samples of P. reticulatum from the Upper Paraguay Basin. The loci were highly polymorphic (mean = 7.28). According to all analysis, the two samples of P. reticulatum revealed pronounced genetic differentiation. Fst value was 0.2290, Rst value 0.1067 and AMOVA 22.90% (Fst ) and 10.67% (Rst ), all being highly significant (p < 0.001). The division of the fishes into two groups was confirmed by microsatellite multi-locus Bayesian assignment testing. The results obtained present evidence of genetic structuring in a P. reticulatum population

Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes

Background: Different moderrn methodologies are presently available to analyze meiotic chromosomes. These methods permit investigation of the behavior of chromosomes in the normal complement and of sex and B chromosomes, two special types of chromosomes that are associated with the A complement and are present in many organisms, including fishes. However, meiotic studies are still scarce in fishes, considering the wide number of species in this group.. Here, we describe a new protocol for the visualization of the synaptonemal complex in spermatocytes and oocytes of fishes and to the sequential use of the technique with other procedures and techniques such as immunodetection of the synaptonemal complex protein with a specific antibody and co-detection of DNA sequences by FISH.Results: The meiotic surface-spreading protocol used in the present proposal worked well in representative species of four fish orders and was useful in obtaining good results even in small specimens. Fish-specific antibodies and commercial products worked similarly well to detect synaptonemal complex (SC) proteins. The sequential application of fluorescence in situ hybridization using specific probes showed clear signals associated with the SC structures identified by immunostaining.Conclusion: Here, we provide a useful and applicable immunofluorescent protocol for the visualization of synaptonemal complex proteins in the meiotic cells of fishes in surface-spreading preparations. Furthermore, this technique allows for the sequential application of other cytogenetic procedures.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Cytogenetic markers as diagnoses in the identification of the hybrid between Piauçu (Leporinus macrocephalus) and Piapara (Leporinus elongatus)

The genetic monitoring of interspecific hybrids involves the application of methodologies able to provide an easy and indubitable genetic characterization of both parental and hybrid individuals. In the present work, cytogenetic techniques were used to identify a hybrid lineage of Piaupara in order to caracterize them in relation to the parental species, Leporinus macrocephalus (piauçu) and L. elongatus (piapara). The cytogenetic analysis revealed that L. macrocephalus presented 2n = 54 chromosomes and a nucleolar organizer regions (NOR) at the telomere of the long arm of the submetacentric chromosome pair 2. Analysis of constitutive heterochromatin (C-banding) revealed a conspicuous block at the pericentromeric region on the long arm of a submetacentric chromosome pair. L. elongatus presented the same diploid number, 2n = 54, and a karyotypic formula similar to that of L. macrocephalus. The NORs were also at the telomere of the long arm of the submetacentric pair 2, which was morphologically different from that of L. macrocephalus. Heterochromatic blocks were observed at both telomeres of a submetacentric chromosome pair. The hybrid Piaupara presented the same diploid number (2n = 54) and karyotypic formula as the parental species and there were no visible differences between parental and hybrid individuals. Differently from the Giemsa staining, NOR- and C-banding analysis showed marked differences which allowed the identification of the hybrids by the different morphology and/or size of the chromosomes carrying the NORs and patterns of heterochromatin distribution in their chromosomes. Such genetic studies are important for fish culture since they can provide tools for monitoring natural and artificial hybridization. They are also useful in biological conservation programmes and in the proper management of natural and reared fish stocks.195202Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Consolidation of the genetic and cytogenetic maps of turbot (Scophthalmus maximus) using FISH with BAC clones

This is a post-peer-review, pre-copyedit version of an article published in Chromosoma. The final authenticated version is available online at: https://doi.org/10.1007/s00412-014-0452-2Bacterial artificial chromosomes (BAC) have been widely used for fluorescence in situ hybridization (FISH) mapping of chromosome landmarks in different organisms, including a few in teleosts. In this study, we used BAC-FISH to consolidate the previous genetic and cytogenetic maps of the turbot (Scophthalmus maximus), a commercially important pleuronectiform. The maps consisted of 24 linkage groups (LGs) but only 22 chromosomes. All turbot LGs were assigned to specific chromosomes using BAC probes obtained from a turbot 5× genomic BAC library. It consisted of 46,080 clones with inserts of at least 100 kb and <5 % empty vectors. These BAC probes contained gene-derived or anonymous markers, most of them linked to quantitative trait loci (QTL) related to productive traits. BAC clones were mapped by FISH to unique marker-specific chromosomal positions, which showed a notable concordance with previous genetic mapping data. The two metacentric pairs were cytogenetically assigned to LG2 and LG16, and the nucleolar organizer region (NOR)-bearing pair was assigned to LG15. Double-color FISH assays enabled the consolidation of the turbot genetic map into 22 linkage groups by merging LG8 with LG18 and LG21 with LG24. In this work, a first-generation probe panel of BAC clones anchored to the turbot linkage and cytogenetical map was developed. It is a useful tool for chromosome traceability in turbot, but also relevant in the context of pleuronectiform karyotypes, which often show small hardly identifiable chromosomes. This panel will also be valuable for further integrative genomics of turbot within Pleuronectiformes and teleosts, especially for fine QTL mapping for aquaculture traits, comparative genomics, and whole-genome assemblyThis study was supported by Spain’s Ministerio de Ciencia e Innovación (AGL2009-13273), Consolider Ingenio Aquagenomics (CSD200700002) and Xunta de Galicia (09MMA011261PR; 10MMA200027PR). Samples for cytogenetic analysis were kindly supplied by Cluster de Acuicultura de GaliciaS