Eugene Yu. Krysanov

Metal–organic frameworks (MOFs) possess remarkable physicochemical properties, making them highly attractive candidates for advanced drug delivery, catalysis, water purification, gas separation, and biomedical application. Thus, it is important to understand its toxic effect on the environment and human health prior to wide practical application. In this study, UiO-66 nanoparticles (Zr₆O₄(OH)₄—octahedral clusters with benzene-1,4-dicarboxylate linkers) were synthesized and characterized by XRD, SEM, and TEM techniques. The genotoxicity and accumulation of UiO-66 were investigated towards freshwater fish, Nothobranchius rachovii, for the first time. To gain a better understanding of the effects of UiO-66 nanoparticles towards N. rachovii, fish were exposed to UiO-66, the linker, and zirconium salts. The genotoxic risk caused by UiO-66 was detected by analyzing the Chromosome Aberrations (CAs), frequencies of Sister Chromatid Exchanges (SCEs), mitotic index, and accumulations of Zr in tissues of fish treated with UiO-66 samples, linker, zirconium salt, and non-exposed control subjects.

During meiosis, homologous chromosomes pair to form synaptonemal complexes (SCs) and exchange genetic material through a process known as meiotic recombination. First, programmed DNA double-strand breaks form, followed by the assembly of recombination foci on SCs. These foci mark the sites of recombination intermediates and future crossovers. Distributions of recombination foci along SCs have been studied in many eukaryotes, revealing the interplay between recombination patterns and genome evolution. However, in fish, data on recombination patterns are scarce, and, for the majority of groups, completely absent. Here, we measure the positions of MLH1 foci in 3,504 SCs from 219 male meiotic cells of an African annual killifish Nothobranchius virgatus, a representative of a genus with remarkable karyotype and genome diversity, and present a detailed statistical analysis of its recombination patterns. We found that, in contrast to the several other fish species characterised to date, recombination in N. virgatus occurs across almost entire chromosome arms, excluding (peri)centromeres and telomeres. In the longest SCs, we observed a proximal and a distal peak of the recombination focus frequency and explained the peaks by chromosome pairing dynamics. We also revealed the typical positions of focus pairs, demonstrated interference between foci, with the minimal interfocus distance of 4 μm, and described regions of the total recombination suppression near centromeres and telomeres. In sum, our study provides a detailed analysis of recombination patterns in a killifish with a fully acrocentric karyotype and contributes to cytogenomic and statistical methodology for future exploration of meiotic recombination patterns.

Sex chromosomes have evolved repeatedly across eukaryotes. The emergence of a sex‐determining (SD) locus is expected to progressively restrict recombination, driving convergent molecular differentiation. However, evidence from taxa like teleost fishes, representing over half of vertebrate species with unmatched diversity in SD systems, challenges this model. Teleost sex chromosomes are often difficult to detect as they experience frequent turnovers, resetting the differentiation process. Nothobranchius killifishes, which include the XY system shared by N. furzeri and N. kadleci and X₁X₂Y systems in six other species, offer a valuable model to study sex chromosome turnovers. We characterised X₁X₂Y systems in five killifish species and found that sex chromosomes evolved at least four times independently. Sex‐determining regions resided near centromeres or predicted chromosome rearrangement breakpoints in N. brieni and N. guentheri, suggesting recombination cold spots may facilitate sex chromosome evolution. Chromosomes representing the XY system in N. furzeri/N. kadleci were sex‐linked also in the outgroup Fundulosoma thierryi, with several genes, including gdf6, residing in the region of differentiation. Although the X₁X₂Y systems of N. guentheri, N. lourensi (both Coastal clade), and N. brieni (Kalahari clade) involved different chromosomes, they shared a potential SD region. We uncovered two sex‐linked evolutionary strata of distinct age in N. guentheri. However, its potential SD gene amhr2 was located in the younger stratum and is hence unlikely to be the ancestral SD gene in this lineage. Our findings suggest recombination landscapes shape sex chromosome turnover and that certain synteny blocks are repeatedly co‐opted as sex chromosomes in killifishes.

Sex chromosomes of teleost fishes often have low levels of differentiation and undergo frequent turnovers. Annual Nothobranchius killifishes comprise representatives with male-heterogametic XY or X1X2Y sex chromosome systems, scattered across their phylogeny, nested within species lacking cytologically detectable sex chromosomes. They thus provide a suitable system to study sex chromosome evolution and turnover. Here, we combined molecular cytogenetics and genomic analyses to examine several multiple sex chromosome systems in Nothobranchius spp. and their outgroup Fundulosoma thierryi. We used fluorescence in situ hybridization with three sex chromosome-specific painting probes and bacterial artificial chromosomes (BAC) bearing eight orthologues of genes found to be repeatedly co-opted as master sex determining (MSD) genes in fishes. Our results suggest at least four independent origins of sex chromosomes in the genus Nothobranchius. The synteny block carrying amhr2 gene was shared by X1X2Y systems of N. brieni, N. guentheri and N. lourensi, but the autosomal additions and the overall neo-Y chromosome structure differed among these species. On the other hand, gdf6 gene was localized to neo-Y of F. thierryi. None of the mapped MSD gene candidates seems to determine sex in N. ditte. We further sequenced genomes of F. thierryi female and N. guentheri male by long-read platforms and performed analyses of male and female Pool-seq data and coverage to delimit their non-recombining regions, determine degree of their differentiation, and thus complement the cytogenetic data in assessing potential MSD genes. We found low level of sex chromosomes differentiation in F. thierryi. In N. guentheri, however, we identified two distinct evolutionary strata on neo-Y. The amhr2 gene resides in the younger stratum and has low allelic variation, which questions its role in sex determination.

Using African annual killifishes of the genus Nothobranchius from temporary savannah pools with rapid karyotype and sex chromosome evolution, we analysed the chromosomal distribution of telomeric (TTAGGG)_n repeat and Nfu‐SatC satellite DNA (satDNA; isolated from Nothobranchius furzeri) in 15 species across the Nothobranchius killifish phylogeny, and with Fundulosoma thierryi as an out‐group. Our fluorescence in situ hybridization experiments revealed that all analysed taxa share the presence of Nfu‐SatC repeat but with diverse organization and distribution on chromosomes. Nfu‐SatC landscape was similar in conspecific populations of Nothobranchius guentheri and Nothobranchius melanospilus but slightly‐to‐moderately differed between populations of Nothobranchius pienaari, and between closely related Nothobranchius kuhntae and Nothobranchius orthonotus. Inter‐individual variability in Nfu‐SatC patterns was found in N. orthonotus and Nothobranchius krysanovi. We revealed mostly no sex‐linked patterns of studied repetitive DNA distribution. Only in Nothobranchius brieni, possessing multiple sex chromosomes, Nfu‐SatC repeat occupied a substantial portion of the neo‐Y chromosome, similarly as formerly found in the XY sex chromosome system of turquoise killifish N. furzeri and its sister species Nothobranchius kadleci—representatives not closely related to N. brieni. All studied species further shared patterns of expected telomeric repeats at the ends of all chromosomes and no additional interstitial telomeric sites. In summary, we revealed (i) the presence of conserved satDNA class in Nothobranchius clades (a rare pattern among ray‐finned fishes); (ii) independent trajectories of Nothobranchius sex chromosome differentiation, with recurrent and convergent accumulation of Nfu‐SatC on the Y chromosome in some species; and (iii) genus‐wide shared tendency to loss of telomeric repeats during interchromosomal rearrangements. Collectively, our findings advance our understanding of genome structure, mechanisms of karyotype reshuffling, and sex chromosome differentiation in Nothobranchius killifishes from the genus‐wide perspective.

The article discusses possible approaches to cytogenetic homeostasis assessment in natural populations. Various cytogenetic methods, their advantages, disadvantages, and limitations related to the study of animals in their natural habitat are considered. For a reliable characterization of environmental quality, it is recommended to supplement the data obtained in natural populations with laboratory experiments using a test object. Due to a number of features, it is proposed to use the short-living fish Nothobranchius rachovii.

Cytogenetic Homeostasis and Environmental Health (Assessment Practice) Цитогенетический гомеостаз и здоровье среды (практика оценки)

Repetitive DNA may have significant impact on genome evolution. African annual killifishes of the genus Nothobranchius (Teleostei: Nothobranchiidae), which has adapted to temporary water pools in African savannahs, possess genomes with high repeat content. They are also characterized by rapid karyotype and sex chromosome evolution but the role of genome repeats in these processes remains largely unknown. Here, we analyzed the distribution of telomeric (TTAGGG) _n repeat and Nfu-SatC satellite DNA (isolated formerly from N. furzeri ) by fluorescence in situ hybridization in representatives across the Nothobranchius phylogeny (15 species), and with Fundulosoma thierryi as an outgroup. All analyzed taxa shared the presence of Nfu-SatC repeat but with diverse organization and distribution on chromosomes (from small clusters scattered genome-wide, to large localized accumulations, or a combined pattern). Nfu-SatC landscape was similar in conspecific populations of N. guentheri and N. melanospilus but slightly-to-moderately differed between populations of N. pienaari , and between closely related N. kuhntae and N. orthonotus . Inter-individual variability in Nfu-SatC patterns was found in N. orthonotus and N. krysanovi , including distinct segments present often in heterozygous condition. We revealed mostly no sex-linked patterns of studied repeat’s distribution in any of the sampled species including those with known sex chromosomes. Only in N. brieni (having an X ₁ X ₂ Y multiple sex chromosome system), Nfu-SatC probe covered substantial portion of the Y chromosome, similarly as formerly found in N. furzeri and N. kadleci (XY sex chromosomes), sister species not closely related to N. brieni . All studied species further shared patterns of telomeric FISH, with expected signals at the ends of all chromosomes and no additional interstitial telomeric sites. In summary, we revealed i) the presence of conserved satDNA class in Nothobranchius clade (a rare pattern among ray-finned fishes), ii) independent trajectories of Nothobranchius sex chromosome diferentiation, with recurrent and convergent accumulation of Nfu-SatC on the Y chromosome in some species, and iii) genus-wide shared propensity to loss of telomeric repeats during the mechanism of interchromosomal rearrangements. Collectively, our findings advance our understanding of genome structure, mechanisms of karyotype reshuffling and sex chromosome differentiation in Nothobranchius killifishes from the genus-wide perspective.

The karyotype differentiation of the twelve known members of the Nothobranchius ugandensis Wildekamp, 1994 species group is reviewed and the karyotype composition of seven of its species is described herein for the first time using a conventional cytogenetic protocol. Changes in the architecture of eukaryotic genomes often have a major impact on processes underlying reproductive isolation, adaptation and diversification. African annual killifishes of the genus Nothobranchius Peters, 1868 (Teleostei: Nothobranchiidae), which are adapted to an extreme environment of ephemeral wetland pools in African savannahs, feature extensive karyotype evolution in small, isolated populations and thus are suitable models for studying the interplay between karyotype change and species evolution. The present investigation reveals a highly conserved diploid chromosome number (2n = 36) but a variable number of chromosomal arms (46–64) among members of the N. ugandensis species group, implying a significant role of pericentric inversions and/or other types of centromeric shift in the karyotype evolution of the group. When superimposed onto a phylogenetic tree based on molecular analyses of two mitochondrial genes the cytogenetic characteristics did not show any correlation with the phylogenetic relationships within the lineage. While karyotypes of many other Nothobranchius spp. studied to date diversified mainly via chromosome fusions and fissions, the N. ugandensis species group maintains stable 2n and the karyotype differentiation seems to be constrained to intrachromosomal rearrangements. Possible reasons for this difference in the trajectory of karyotype differentiation are discussed. While genetic drift seems to be a major factor in the fixation of chromosome rearrangements in Nothobranchius, future studies are needed to assess the impact of predicted multiple inversions on the genome evolution and species diversification within the N. ugandensis species group.

Surface modification of nanoparticles with different stabilizers is one of the most widely used methods to improve their stability and applicability. Silver nanoparticle (AgNPs) dispersions with biologically active stabilizers have great potential as plant protection products with synergetic antimicrobial properties and sufficient stability in terms of field application. The obtained AgNPs dispersions have the ability to enhance growth, increase yield and give better protection to various crops. At the same time, it is important to determine the fate, stability, and ecotoxicity of the applied nanosized products. The toxic effects of AgNPs dispersions and their constituents, organic stabilizers and additives, were evaluated using a phenotypic sea urchin embryo assay. Certain AgNPs dispersions with organic stabilizers demonstrated sufficient stability, even in seawater. The toxicity of the AgNPs decreased with the increasing tendency to agglomerate in seawater. Furthermore, the applied stabilizers were hazardous towards sea urchin embryos. They caused pronounced embryo abnormalities at 0.25–2.6 mg/L concentrations. AgNPs exhibited a lethal effect at concentrations that were equal to the MLC or exceeded the MEC of their stabilizers. Silver ions were more toxic towards sea urchin embryos than AgNPs.