Hybridization of Russian Sturgeon (Acipenser gueldenstaedtii, Brandt and Ratzeberg, 1833) and American Paddlefish (Polyodon spathula, Walbaum 1792) and Evaluation of Their Progeny.

Two species from the families Acipenseridae and Polyodontidae, Russian sturgeon (Acipenser gueldenstaedtii, Brandt and Ratzeberg, 1833; functional tetraploid) and American paddlefish (Polyodon spathula, Walbaum 1792, functional diploid) were hybridized. The hybridization was repeated using eggs from three sturgeon and sperm from four paddlefish individuals. Survival in all hybrid family groups ranged from 62% to 74% 30 days after hatching. This was the first successful hybridization between these two species and between members of the family Acipenseridae and Polyodontidae. Flow cytometry and chromosome analysis revealed two ploidy levels in hybrids. The chromosome numbers of the hybrids ranged between 156-184 and 300-310, in "functional" triploids and "functional" pentaploids, respectively. The hybrid origin and the ploidy levels were also confirmed by microsatellite analyses. In hybrids, the size and the number of dorsal and ventral scutes correlated with the ploidy levels as well as with the calculated ratio of the maternal and paternal chromosome sets. An extra haploid cell lineage was found in three hybrid individuals irrespective of the ploidy level, suggesting polyspermy. Although the growth performance showed high variance in hybrids (mean: 1.2 kg, SD: 0.55), many individuals reached a size of approximately 1 kg by the age of one year under intensive rearing conditions.

Effects of high ambient temperature on fish sperm plasma membrane integrity and mitochondrial activity - A flow cytometric study.

Local extreme climatic conditions occurring as a result of global climate change may interfere with the reproduction of animals. In the present study fish spermatozoa were incubated at different temperatures (20, 25, 30 and 40 °C) for 10 and 30 minutes, respectively and plasma membrane integrity and mitochondrial membrane potential changes were evaluated with flow cytometry using SYBR-14/PI and Mitotracker Deep Red FM fluorescent dyes. No significant differences were found in plasma membrane integrity at either incubation temperatures or time points. Mitotracker Deep Red FM histogram profiles indicating mitochondrial activity showed significant (p < 0.001) alterations in all cases of higher (25, 30 and 40 °C) temperature treatments as compared to the samples incubated at 20 °C. Our results indicate that fish spermatozoa exposed to high temperatures suffer sublethal damage that cannot be detected with conventional, vital staining techniques.

Flow cytometric detection of oxidative DNA damage in fish spermatozoa exposed to cadmium - Short communication.

The aim of the present pilot study was to apply a flow cytometric assay, the so-called OxyDNA test, to determine the level of oxidative DNA damage in fish spermatozoa exposed to different concentrations (0.01-10,000 mg/L) of cadmium. Milt was collected from three randomly selected Prussian carp (Carassius auratus gibelio) males. Oxidative DNA damage was assessed with the OxyDNA kit and using flow cytometry. The ratio of OxyDNA-positive events increased significantly at higher cadmium concentrations. The results indicate that direct contact of fish spermatozoa with cadmium-polluted water initiates genotoxic damage.

The complete mitochondrial genome of perch Perca fluviatilis (Perciformes: Percidae).

In this study, we cloned and sequenced the complete mitochondrial genome of Perca fluviatilis. It was a circular double-stranded DNA molecule of 16,537 base pairs (bp) in length and consists of 13 protein-coding genes, 22 tRNA genes, two ribosomal RNA genes and two main non-coding regions (the control region and the origin of the light strand replication). The mitogenome of shared common features with those of other toleosts in terms of gene order and base composition. All protein-coding genes were initiated with ATG except for COX 1, which began with GTG instead. However, the termination codons of 13 protein-coding genes are varied with TAG, TAA or T. This mitogenome sequence data would contribute to better understanding phylogenetic relationships and population genetics of the family Percidae.

The complete mitochondrial genome of pike-perch, Sander lucioperca (Perciformes: Percidae).

The complete mitochondrial genome of Sander lucioperca has been sequenced and analyzed in this study. It was a circular double-stranded DNA molecule of 16,541 base pairs (bp) in length and exhibited 37 typical animal mitochondrial genes. The gene order and base composition were similar to those of other percid species. All protein-coding genes were initiated with ATG except for COX 1, which began with GTG instead. However, the termination codons of 13 protein-coding genes varied with TAG, TAA, TA or T. Within CR, we detected five copies of 10 bp tandemly repeated sequences domain, which immediately followed the tRNA(Pro). These mitogenome sequence data would contribute to better understanding phylogenetic relationships and population genetics of the family Percidae.

Comparative Mitogenomics of the Genus Odontobutis (Perciformes: Gobioidei: Odontobutidae) Revealed Conserved Gene Rearrangement and High Sequence Variations.

To understand the molecular evolution of mitochondrial genomes (mitogenomes) in the genus Odontobutis, the mitogenome of Odontobutis yaluensis was sequenced and compared with those of another four Odontobutis species. Our results displayed similar mitogenome features among species in genome organization, base composition, codon usage, and gene rearrangement. The identical gene rearrangement of trnS-trnL-trnH tRNA cluster observed in mitogenomes of these five closely related freshwater sleepers suggests that this unique gene order is conserved within Odontobutis. Additionally, the present gene order and the positions of associated intergenic spacers of these Odontobutis mitogenomes indicate that this unusual gene rearrangement results from tandem duplication and random loss of large-scale gene regions. Moreover, these mitogenomes exhibit a high level of sequence variation, mainly due to the differences of corresponding intergenic sequences in gene rearrangement regions and the heterogeneity of tandem repeats in the control regions. Phylogenetic analyses support Odontobutis species with shared gene rearrangement forming a monophyletic group, and the interspecific phylogenetic relationships are associated with structural differences among their mitogenomes. The present study contributes to understanding the evolutionary patterns of Odontobutidae species.

Disappearing scales in carps: re-visiting Kirpichnikov's model on the genetics of scale pattern formation.

The body of most fishes is fully covered by scales that typically form tight, partially overlapping rows. While some of the genes controlling the formation and growth of fish scales have been studied, very little is known about the genetic mechanisms regulating scale pattern formation. Although the existence of two genes with two pairs of alleles (S&s and N&n) regulating scale coverage in cyprinids has been predicted by Kirpichnikov and colleagues nearly eighty years ago, their identity was unknown until recently. In 2009, the 'S' gene was found to be a paralog of fibroblast growth factor receptor 1, fgfr1a1, while the second gene called 'N' has not yet been identified. We re-visited the original model of Kirpichnikov that proposed four major scale pattern types and observed a high degree of variation within the so-called scattered phenotype due to which this group was divided into two sub-types: classical mirror and irregular. We also analyzed the survival rates of offspring groups and found a distinct difference between Asian and European crosses. Whereas nude × nude crosses involving at least one parent of Asian origin or hybrid with Asian parent(s) showed the 25% early lethality predicted by Kirpichnikov (due to the lethality of the NN genotype), those with two Hungarian nude parents did not. We further extended Kirpichnikov's work by correlating changes in phenotype (scale-pattern) to the deformations of fins and losses of pharyngeal teeth. We observed phenotypic changes which were not restricted to nudes, as described by Kirpichnikov, but were also present in mirrors (and presumably in linears as well; not analyzed in detail here). We propose that the gradation of phenotypes observed within the scattered group is caused by a gradually decreasing level of signaling (a dose-dependent effect) probably due to a concerted action of multiple pathways involved in scale formation.

Duplication of fgfr1 permits Fgf signaling to serve as a target for selection during domestication.

The genetic basis of morphological variation both within and between species has been a lasting question in evolutionary biology and one of considerable recent debate. It is thought that changes in postembryonic development leading to variations in adult form often serve as a basis for selection . Thus, we investigated the genetic basis of the development of adult structures in the zebrafish via a forward genetic approach and asked whether the genes and mechanisms found could be predictive of changes in other species. Here we describe the spiegeldanio (spd) zebrafish mutation, which leads to reduced scale formation in the adult. The affected gene is fibroblast growth factor receptor 1 (fgfr1), which is known to have an essential embryonic function in vertebrate development. We find that the zebrafish has two paralogs encoding Fgfr1 and show that they function redundantly during embryogenesis. However, only one paralog is required for formation of scales during juvenile development. Furthermore, we identify loss-of-function alleles changing the coding sequence of Fgfr1a1 that have been independently selected twice during the domestication of the carp (Cyprinus carpio). These findings provide evidence for the role for gene duplication in providing the raw material for generation of morphological diversity.