Chromosomal Evolution and Evolutionary Relationships of Lebiasina Species (Characiformes, Lebiasinidae)

We present the first cytogenetic data for Lebiasina bimaculata and L. melanoguttata with the aim of (1) investigating evolutionary events within Lebiasina and their relationships with other Lebiasinidae genera and (2) checking the evolutionary relationships between Lebiasinidae and Ctenoluciidae. Both species have a diploid number 2n = 36 with similar karyotypes and microsatellite distribution patterns but present contrasting C-positive heterochromatin and CMA3 + banding patterns. The remarkable interstitial series of C-positive heterochromatin occurring in L. melanoguttata is absent in L. bimaculata. Accordingly, L. bimaculata shows the ribosomal DNA sites as the only GC-rich (CMA3 +) regions, while L. melanoguttata shows evidence of a clear intercalated CMA3 + banding pattern. In addition, the multiple 5S and 18S rDNA sites in L. melanogutatta contrast with single sites present in L. bimaculata. Comparative genomic hybridization (CGH) experiments also revealed a high level of genomic differentiation between both species. A polymorphic state of a conspicuous C-positive, CMA3 +, and (CGG)n band was found only to occur in L. bimaculata females, and its possible relationship with a nascent sex chromosome system is discussed. Whole chromosome painting (WCP) and CGH experiments indicate that the Lebiasina species examined and Boulengerella maculata share similar chromosomal sequences, thus supporting the relatedness between them and the evolutionary relationships between the Lebiasinidae and Ctenoluciidae families

Cytogenetics, genomics and biodiversity of the South American and African Arapaimidae fish family (Teleostei, Osteoglossiformes)

<div><p>Osteoglossiformes represents one of the most ancestral teleost lineages, currently widespread over almost all continents, except for Antarctica. However, data involving advanced molecular cytogenetics or comparative genomics are yet largely limited for this fish group. Therefore, the present investigations focus on the osteoglossiform family Arapaimidae, studying a unique fish model group with advanced molecular cytogenetic genomic tools. The aim is to better explore and clarify certain events and factors that had impact on evolutionary history of this fish group. For that, both South American and African representatives of Arapaimidae, namely <i>Arapaima gigas</i> and <i>Heterotis niloticus</i>, were examined. Both species differed markedly by diploid chromosome numbers, with 2n = 56 found in <i>A</i>. <i>gigas</i> and 2n = 40 exhibited by <i>H</i>. <i>niloticus</i>. Conventional cytogenetics along with fluorescence <i>in situ</i> hybridization revealed some general trends shared by most osteoglossiform species analyzed thus far, such as the presence of only one chromosome pair bearing 18S and 5S rDNA sites and karyotypes dominated by acrocentric chromosomes, resembling thus the patterns of hypothetical ancestral teleost karyotype. Furthermore, the genomes of <i>A</i>. <i>gigas</i> and <i>H</i>. <i>niloticus</i> display remarkable divergence in terms of repetitive DNA content and distribution, as revealed by comparative genomic hybridization (CGH). On the other hand, genomic diversity of single copy sequences studied through principal component analyses (PCA) based on SNP alleles genotyped by the DArT seq procedure demonstrated a very low genetic distance between the South American and African Arapaimidae species; this pattern contrasts sharply with the scenario found in other osteoglossiform species. Underlying evolutionary mechanisms potentially explaining the obtained data have been suggested and discussed.</p></div

Centric fusions behind the karyotype evolution of neotropical nannostomus pencilfishes (Characiforme, Lebiasinidae): First insights from a molecular cytogenetic perspective

Lebiasinidae is a Neotropical freshwater family widely distributed throughout South and Central America. Due to their often very small body size, Lebiasinidae species are cytogenetically challenging and hence largely underexplored. However, the available but limited karyotype data already suggested a high interspecific variability in the diploid chromosome number (2n), which is pronounced in the speciose genus Nannostomus, a popular taxon in ornamental fish trade due to its remarkable body coloration. Aiming to more deeply examine the karyotype diversification in Nannostomus, we combined conventional cytogenetics (Giemsa-staining and C-banding) with the chromosomal mapping of tandemly repeated 5S and 18S rDNA clusters and with interspecific comparative genomic hybridization (CGH) to investigate genomes of four representative Nannostomus species: N. beckfordi, N. eques, N. marginatus, and N. unifasciatus. Our data showed a remarkable variability in 2n, ranging from 2n = 22 in N. unifasciatus (karyotype composed exclusively of metacentrics/submetacentrics) to 2n = 44 in N. beckfordi (karyotype composed entirely of acrocentrics). On the other hand, patterns of 18S and 5S rDNA distribution in the analyzed karyotypes remained rather conservative, with only two 18S and two to four 5S rDNA sites. In view of the mostly unchanged number of chromosome arms (FN = 44) in all but one species (N. eques; FN = 36), and with respect to the current phylogenetic hypothesis, we propose Robertsonian translocations to be a significant contributor to the karyotype differentiation in (at least herein studied) Nannostomus species. Interspecific comparative genome hybridization (CGH) using whole genomic DNAs mapped against the chromosome background of N. beckfordi found a moderate divergence in the repetitive DNA content among the species’ genomes. Collectively, our data suggest that the karyotype differentiation in Nannostomus has been largely driven by major structural rearrangements, accompanied by only low to moderate dynamics of repetitive DNA at the sub-chromosomal level. Possible mechanisms and factors behind the elevated tolerance to such a rate of karyotype change in Nannostomus are discussed. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Cytogenetics of the small-sized fish, Copeina guttata (Characiformes, Lebiasinidae): Novel insights into the karyotype differentiation of the family

Lebiasinidae is a small fish family composed by miniature to small-sized fishes with few cytogenetic data (most of them limited to descriptions of diploid chromosome numbers), thus preventing any evolutionary comparative studies at the chromosomal level. In the present study, we are providing, the first cytogenetic data for the red spotted tetra, Copeina guttata, including the standard karyotype, C-banding, repetitive DNA mapping by fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH), providing chromosomal patterns and novel insights into the karyotype differentiation of the family. Males and females share diploid chromosome number 2n = 42 and karyotype composed of 2 metacentric (m), 4 submetacentric (sm) and 36 subtelocentric to acrocentric (st-a) chromosomes. Blocks of constitutive heterochromatin were observed in the centromeric and interstitial regions of several chromosomes, in addition to a remarkably large distal block, heteromorphic in size, which fully corresponded with the 18S rDNA sites in the fourth chromosomal pair. This overlap was confirmed by 5S/18S rDNA dual-color FISH. On the other hand, 5S rDNA clusters were situated in the long and short arms of the 2nd and 15th pairs, respectively. No sex-linked karyotype differences were revealed by male/female CGH experiments. The genomic probes from other two lebiasinid species, Lebiasina melanoguttata and Pyrrhulina brevis, showed positive hybridization signals only in the NOR region in the genome of C. guttata. We demonstrated that karyotype diversification in lebiasinids was accompanied by a series of structural and numeric chromosome rearrangements of different types, including particularly fusions and fissions. © 2019 Toma et al

An Insight into the Chromosomal Evolution of Lebiasinidae (Teleostei, Characiformes)

Lebiasinidae fishes have been historically neglected by cytogenetical studies. Here we present a genomic comparison in eleven Lebiasinidae species, in addition to a review of the ribosomal DNA sequences distribution in this family. With that, we develop ten sets of experiments in order to hybridize the genomic DNA of representative species from the genus Copeina, Copella, Nannostomus, and Pyrrhulina in metaphase plates of Lebiasina melanoguttata. Two major pathways on the chromosomal evolution of these species can be recognized: (i) conservation of 2n = 36 bi-armed chromosomes in Lebiasininae, as a basal condition, and (ii) high numeric and structural chromosomal rearrangements in Pyrrhulininae, with a notable tendency towards acrocentrization. The ribosomal DNA (rDNA) distribution also revealed a marked differentiation during the chromosomal evolution of Lebiasinidae, since both single and multiple sites, in addition to a wide range of chromosomal locations can be found. With some few exceptions, the terminal position of 18S rDNA appears as a common feature in Lebiasinidae-analyzed species. Altogether with Ctenoluciidae, this pattern can be considered a symplesiomorphism for both families. In addition to the specific repetitive DNA content that characterizes the genome of each particular species, Lebiasina also keeps inter-specific repetitive sequences, thus reinforcing its proposed basal condition in Lebiasinidae

RAPD markers indicate the occurrence of structured populations in a migratory freshwater fish species

Many factors have contributed to the destruction of fish habitats. Hydroelectric dams, water pollution and other environmental changes have resulted in the eradication of natural stocks. The aim of this study was to detect the genetic variation in Prochilodus marggravii from three collection sites in the area of influence of the Três Marias dam (MG) on the São Francisco river (Brazil), using the RAPD technique. The results obtained revealed that the fish in the downstream region nearest the dam have a higher similarity coefficient than those from the other sampling sites that may be related to differences in environmental characteristics in these regions. Additionaly, significant differences in the band frequencies were observed from one collection site to another. These both findings suggest the occurrence of a structured population and have important implications for the conservation of the genetic variability of distinct natural P. marggravii stocks

Heterochromatin heterogeneity and chromosome heteromorphism in Cerdocyon thous (Mammalia, Canidae)

Few cytogenetic studies have been conducted on South American canids. Cerdocyon thous is a representative of the Canidae, living in the forests, open wooded areas and savannahs of South America. Compared to other canid species, C. thous has a large proportion of metacentric and submetacentric chromosomes as well as a large amount of constitutive heterochromatin, especially along the short arm of submetacentric chromosomes. In the present study, different chromosome banding methods were used to characterize the heterogeneous nature of the large heterochromatic segments and to propose an organizational model for this segment that occupies the entire short arm of most two-armed chromosomes. Furthermore, chromosome heteromorphism related to the short arm of large submetacentric chromosome corresponding to no. 3 in male and female karyotypes is described.<br>Os estudos citogenéticos em canídeos sul-americanos, de uma forma geral, são raros. Cerdocyon thous é um representante da família Canidae e habita florestas, matas abertas e savanas da américa do Sul. O seu cariótipo é formado por uma grande proporção de cromossomos metacêntricos e submetacêntricos quando comparado com outras espécies desta família e possui também uma grande quantidade de heterocromatina constitutiva, especialmente ao longo do braço curto dos cromossomos submetacêntricos. No presente estudo, diferentes métodos de bandamentos cromossômicos permitiram verificar o caráter heterogêneo dos grandes segmentos heterocromáticos e a proposição de um modelo de organização deste segmento que ocupa todo o braço curto da maioria dos cromossomos de dois braços. Além disso, é descrito um heteromorfismo cromossômico relacionado ao braço curto de um grande cromossomo submetacêntrico, equivalente ao no. 3 do cariótipo de animais de ambos os sexos

Evidence of lack of population substructuring in the Brazilian freshwater fish Prochilodus costatus

Curimbatá-pióa (Prochilodus costatus Valenciennes, 1850) is an endemic species from the São Francisco River Basin with migratory habits, important ecological role in the ecosystem and relevant fishing importance. The present study aimed to assess the genetic variation in P. costatus in order to investigate its population genetic structure. Genetic variation was studied at three sites downstream the Três Marias dam (Upper-middle São Francisco River, State of Minas Gerais) through six specific microsatellite loci. Fish from the three sites had quite similar genetic diversity levels and no genetic differentiation was detected, suggesting that P. costatus might represent a single reproductive unit in the studied area. Alternatively, the present study was not able to detect putative coexistent and comigrating populations along the main channel river. Our findings could be very helpful for the management and conservation of this fish

DNA cloning and expression analysis of the catalytic subunit of glutamate cysteine ligase gene in an annelid polychaete after cadmium exposure: a potential tool for pollution biomonitorin

The enzyme glutamate cysteine ligase (GCL) is the rate-limiting enzyme in glutathione (GSH) synthesis and is formed by a catalytic (GCLC) and a modulatory subunit (GCLM). Some studies have demonstrated that environmental pollutants can regulate the expression of these subunits. Despite the importance of these genes in toxicological responses, no sequences are available for the GCL subunits in annelids. The present study reports, for the first time, the cDNA sequence for the GCLC in an annelid species, the polychaete Laeonereis acuta (Nereididae). The deduced amino acid sequence of L. acuta GCLC showed homology with other animal species, and was used to infer a phylogenetic tree with GCLC amino acid sequences from other taxonomic groups. Exposure to cadmium (100 and 1000 μg Cd/L) during 14 days augmented the level of L. acuta GCLC transcripts in a dose-dependent manner. These gene expression results can be related to the known cadmium effect on GSH depletion.Since a number of contaminants can also exert their toxicity through this mechanism, GCLC gene expression might be applied not only for cadmium biomonitoring, but also for a wide range of contaminants that directly or indirectly promote the same effect in the cellular GSH content