Several species of Tityus (Scorpiones, Buthidae) present multi-chromosomal meiotic associations and failures in the synaptic process, originated from reciprocal translocations. Holocentric ...chromosomes and achiasmatic meiosis in males are present in all members of this genus. In the present study, we investigated synapse dynamics, transcriptional silencing by γH2AX, and meiotic microtubule association in bivalents and a quadrivalent of the scorpion Tityus maranhensis. Additionally, we performed RT-PCR to verify the expression of mismatch repair enzymes involved in crossing-over formation in Tityus silvestris gonads. The quadrivalent association in T. maranhensis showed delay in the synaptic process and long asynaptic regions during pachytene. In this species, γH2AX was recorded only at the chromosome ends during early stages of prophase I; in metaphase I, bivalents and quadrivalents of T. maranhensis exhibited binding to microtubules along their entire length, while in metaphase II/anaphase II transition, spindle fibers interacted only with telomeric regions. Regarding T. silvestris, genes involved in the recombination process were transcribed in ovaries, testes and embryos, without significant difference between these tissues. The expression of these genes during T. silvestris achiasmatic meiosis is discussed in the present study. The absence of meiotic inactivation by γH2AX and holo/telokinetic behavior of the chromosomes are important factors for the maintenance of the quadrivalent in T. maranhensis and the normal continuation of the meiotic cycle in this species.
Heterozygous chromosomal rearrangements can result in failures during the meiotic cycle and the apoptosis of germline, making carrier individuals infertile. The Amazon frog Leptodactylus ...pentadactylus has a meiotic multivalent, composed of 12 sex chromosomes. The mechanisms by which this multi-chromosome system maintains fertility in males of this species remain undetermined. In this study we investigated the meiotic behavior of this multivalent to understand how synapse, recombination and epigenetic modifications contribute to maintaining fertility and chromosomal sexual determination in this species. Our sample had 2n = 22, with a ring formed by ten chromosomes in meiosis, indicating a new system of sex determination for this species (X1Y1X2Y2X3Y3X4Y4X5Y5). Synapsis occurs in the homologous terminal portion of the chromosomes, while part of the heterologous interstitial regions performed synaptic adjustment. The multivalent center remains asynaptic until the end of pachytene, with interlocks, gaps and rich-chromatin in histone H2A phosphorylation at serine 139 (γH2AX), suggesting transcriptional silence. In late pachytene, paired regions show repair of double strand-breaks (DSBs) with RAD51 homolog 1 (Rad51). These findings suggest that Rad51 persistence creates positive feedback at the pachytene checkpoint, allowing meiosis I to progress normally. Additionally, histone H3 trimethylation at lysine 27 in the pericentromeric heterochromatin of this anuran can suppress recombination in this region, preventing failed chromosomal segregation. Taken together, these results indicate that these meiotic adaptations are required for maintenance of fertility in L. pentadactylus.
ABSTRACT
In the Amazon, some species of Loricariidae are at risk of extinction due to habitat loss and overexploitation by the ornamental fish market. Cytogenetic data related to the karyotype and ...meiotic cycle can contribute to understanding the reproductive biology and help management and conservation programs of these fish. Additionally, chromosomal mapping of repetitive DNA in Loricariidae may aid comparative genomic studies in this family. However, cytogenetics analysis is limited in Amazonian locariids. In this study, chromosomal mapping of multigenic families was performed in Scobinancistrus aureatus, Scobinancistrus pariolispos and Spatuloricaria sp. Meiotic analyzes were performed in Hypancistrus zebra and Hypancistrus sp. “pão”. Results showed new karyotype for Spatuloricaria sp. (2n=66, NF=82, 50m-10sm-6m). Distinct patterns of chromosomal organization of histone H1, histone H3 and snDNA U2 genes were registered in the karyotypes of the studied species, proving to be an excellent cytotaxonomic tool. Hypotheses to explain the evolutionary dynamics of these sequences in studied Loricariidae were proposed. Regarding H. zebra and H. sp. “pão”, we describe the events related to synapse and transcriptional activity during the meiotic cycle, which in both species showed 26 fully synapsed bivalents, with high gene expression only during zygotene and pachytene. Both Hypancistrus species could be used may be models for evaluating changes in spermatogenesis of Loricariidae.
Holocentric chromosomes occur in approximately 750 species of eukaryotes. Among them, the genus Tityus (Scorpiones, Buthidae) has a labile karyotype that shows complex multivalent associations during ...male meiosis. Thus, taking advantage of the excellent model provided by the Buthidae scorpions, here we analyzed the chromosomal distribution of several repetitive DNA classes on the holocentric chromosomes of different populations of the species Tityus obscurus Gervais, 1843, highlighting their involvement in the karyotypic differences found among them.
This species shows inter- and intrapopulational karyotype variation, with seven distinct cytotypes: A (2n = 16), B (2n = 14), C (2n = 13), D (2n = 13), E (2n = 12), F (2n = 12) and G (2n = 11). Furthermore, exhibits achiasmatic male meiosis and lacks heteromorphic sex chromosomes. Trivalent and quadrivalent meiotic associations were found in some cytotypes. In them, 45S rDNAs were found in the terminal portions of two pairs, while TTAGG repeats were found only at the end of the chromosomes. In the cytotype A (2n = 16), the U2 snRNA gene mapped to pair 1, while the H3 histone cluster and C
t-1 DNA fraction was terminally distributed on all pairs. Mariner transposons were found throughout the chromosomes, with the exception of one individual of cytotype A (2n = 16), in which it was concentrated in heterochromatic regions.
Chromosomal variability found in T. obscurus are due to rearrangements of the type fusion/fission and reciprocal translocations in heterozygous. These karyotype differences follow a geographical pattern and may be contributing to reproductive isolation between populations analyzed. Our results also demonstrate high mobility of histone H3 genes. In contrast, other multigene families (45S rDNA and U2 snRNA) have conserved distribution among individuals. The accumulation of repetitive sequences in distal regions of T. obscurus chromosomes, suggests that end of chromosome are not covered by the kinetochore.
Achiasmatic male meiosis in scorpions is characterized by a high frequency of gaps, asynaptic regions and multivalent associations. Here, we performed an immunocytogenetic analysis to investigate ...recombination, and synapsis and chromatin-remodeling events during meiosis of the scorpion
Our results demonstrate that the synaptonemal complex (SC) begins its organization in the zygotene stage and persists until metaphase I. The advancement of the synaptic process is related to the epigenetic modification histone H3 lysine 27 trimethylation (H3K27m3). The distribution and dynamics patterns of variant γH2AX and recombinase Rad51 during achiasmatic meiosis suggests formation and repair of DNA double-strand breaks (DSBs) during early stages of prophase I. The epigenetic modifications, histone H3 lysine 4 trimethylation (H3K4m3) and histone H3 lysine 9 acetylation (H3K9ac), showed a dispersed distribution along the bivalents, suggesting that transcriptional activity is maintained constitutively during prophase I. However, H3K9ac modifications are absent in constitutive heterochromatin carrying the 45S rDNA in pachytene and post-pachytene stages. Collectively, our data demonstrate that
exhibits adaptations to the achiasmatic mode, and suggest that epigenetic modifications may act in the regulation of these mechanisms to favor the normal continuation of meiosis in this scorpion.
(Cichliformes, Cichlidae) present a highly conserved diploid number 2n=48 with fundamental numbers varying between 52 and 62. We analyzed four species in order to investigate the role of repetitive ...DNA in chromosome evolution in the genus.
,
cf.
and
cf.
have 2n=48 (8 m/sm and 40st/a) and FN=56, while
sp. 'Xingu I' has 2n=48 (48 st/a) and FN=48. Different patterns of constitutive heterochromatin distribution were observed including pericentric, interstitial and whole arm C bands. A single chromosome bears 18S rDNA clusters in most species, except
, where population variation exists in terms of the quantity and distribution of clusters and their association with interstitial telomeric sequences. All species showed hybridization of 5S rDNA sequences in an interstitial region on an acrocentric chromosome pair. The karyotypic differences and maintenance of the diploid number supports chromosome evolution mediated by inversions in
The telomeric and 18S rDNA sequence association in various chromosomes of
are proposed to represent hotspots for breakage, favoring intra-chromosomal rearrangements. The results suggest that repetitive sequences can contribute to microstructural cytogenetic diversity in
.
ABSTRACT Scorpion envenoming is considered a public health problem in Brazil. A recent study described a variation in the clinical outcome of envenoming by Tityus obscurus in two populations ...separated by 850 km in the northeastern Amazon region. Our aim was to evaluate whether such clinical and toxinological variations are associated with underlying differences in genetic diversity between these two T. obscurus populations. We obtained DNA from five individuals of each population, in the municipalities of Belém and Santarém, located east and west of the state of Pará, Brazil, respectively. Gene regions encoding mitochondrial DNA (mtDNA) markers cytochrome oxidase subunit I (COI) and ribosomal 16S RNA (16S) were amplified and sequenced. Phylogenetic analyses were performed using maximum likelihood (ML) and Bayesian inferences (BA) for both molecular data (COI and 16S). The sampled T. obscurus populations corresponded to two distinct mtDNA lineages (genetic distance COI K2 P = 0.08 to 0.13; 16S K2 P = 0.10 to 0.11) with no shared mutations between groups and well supported by ML and BA inferences. Based on the divergence values found between eastern and western populations (COI, 0.07 to 0.12; 16S, 0.10), our study confirms the genetic heterogeneity of T. obscurus populations within the state of Pará, which correlates with observed venom and clinical differences, and reinforces the need for mapping the distribution of haplotypes throughout the geographic range of T. obscurus, to aid in future epidemiological, toxinological, and evolutionary studies.
RESUMO O envenenamento por escorpiões é considerado um problema de saúde pública no Brasil. Um estudo recente descreve a variação do quadro clínico de envenenamento por Tityus obscurus em duas populações separadas por uma distância de 850 km no nordeste da Amazônia. O objetivo deste estudo foi avaliar se tais variações clínicas e toxicológicas estão associadas a diferenças subjacentes na diversidade genética entre essas duas populações de T. obscurus. Obtivemos DNA de cinco indivíduos de cada população nos municípios de Belém e Santarém, localizados a leste e oeste do estado do Pará, Brasil, respectivamente. Regiões gênicas que codificam marcadores de DNA mitocondrial (mtDNA) citocromo oxidase subunidade I (COI) e RNA ribossômico 16S (16S) foram amplificadas e sequenciadas. As análises filogenéticas foram realizadas por máxima verossimilhança (ML) e inferência Bayesiana (BA) para ambos os dados moleculares (COI e 16S). As populações de T. obscurus amostradas corresponderam a duas linhagens distintas de mtDNA (distância genética COI K2 P = 0,08 a 0,13; 16S K2 P = 0,10 a 0,11) sem mutações compartilhadas entre os grupos, e bem corroboradas por inferências ML e BA. Com base nos valores de divergência encontrados entre as populações oriental e ocidental (COI, 0,07 a 0,12; 16S, 0,10), nosso estudo confirma a heterogeneidade genética das populações de T. obscurus no estado do Pará. Os resultados são congruentes com as diferenças observadas no quadro clínico dos acidentes e toxicidade do veneno, e reforçam a necessidade de mapear a distribuição de haplótipos em toda a distribuição geográfica de T. obscurus, para auxiliar em futuros estudos epidemiológicos, toxinológicos e evolutivos.
Exceptionally well-preserved three-dimensional insects with fine details and even labile tissues are ubiquitous in the Crato Member Konservat Lagerstätte (northeastern Brazil). Here we investigate ...the preservational pathways which yielded such specimens. We employed high resolution techniques (EDXRF, SR-SXS, SEM, EDS, micro Raman, and PIXE) to understand their fossilisation on mineralogical and geochemical grounds. Pseudomorphs of framboidal pyrite, the dominant fossil microfabric, display size variation when comparing cuticle with inner areas or soft tissues, which we interpret as the result of the balance between ion diffusion rates and nucleation rates of pyrite through the originally decaying carcasses. Furthermore, the mineral fabrics are associated with structures that can be the remains of extracellular polymeric substances (EPS). Geochemical data also point to a concentration of Fe, Zn, and Cu in the fossils in comparison to the embedding rock. Therefore, we consider that biofilms of sulphate reducing bacteria (SRB) had a central role in insect decay and mineralisation. Therefore, we shed light on exceptional preservation of fossils by pyritisation in a Cretaceous limestone lacustrine palaeoenvironment.
Loricariidae (Siluriformes) comprises ∼1026 species of neotropical fish, being considered the most diverse among the Siluriformes. Studies on repetitive DNA sequences have provided important data on ...the evolution of the genomes of members of this family, especially of the Hypostominae subfamily. In this study, the chromosomal mapping of the histone multigene family and U2 snRNA was performed in two species belonging to the
genus,
sp. "pão" (2n = 52, 22m + 18sm +12st) and
(2n = 52, 16m + 20sm +16st). The presence of dispersed signals of histones H2A, H2B, H3, and H4 in the karyotype of both species, with each sequence displaying a varied level of accumulation and dispersion of these sequences between them was observed; in addition, U2 snDNA probe only showed positive results in
, which present this multigene in the terminal region of three chromosomal pairs. The obtained results resemble data already analyzed in the literature, in which the action of transposable elements interfere in the organization of these multigene families, in addition to other evolutionary processes that shape the evolution of the genome, such as circular or ectopic recombination. This study also shows that the dispersion of the multigene histone family is quite complex, and from this, these data serve as a point of discussion for the evolutionary processes that occur in the
karyotype.
There are several scorpion species of medical relevance around the world. Some of them are well characterized by their toxins and clinical outcomes. Brazilian Amazon has a great amount of these ...arthropods that have an impact in the scorpionism events specifically in this region of Brazil. Recently, several studies pointed out the immune system activation during scorpion envenouming as an important facet of scorpionism, inducing a sepsis-like state that culminates in clinical severity and death. In this work, we characterized the macrophage response of three species of clinical relevance in Brazilian Amazon: Tityus silvestris, T. metuendus and T. obscurus and one specie with no toxic effects to humans, Brotheas amazonicus. All the four species analyzed were able to induce pro- and anti-inflammatory cytokine production in a J774.1 murine macrophage model. This activation was dependent on TLR2/TLR4/MyD88 activation and abolished by TLRs antagonists. These results suggest that the venoms of the four species analyzed were able to induce macrophage response in agreement to the well-established immune activation by T. serrulatus venom. Our findings provide new insights into the clinical repercussions of scorpionism of uncharacterized species and point to new biotechnological applications of these venoms and possible supportive therapies in scorpionism.
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•Scorpion Venoms from the Brazilian Amazon induce inflammatory macrophage response.•Scorpion venoms-stimulated macrophages release pro and anti-inflammatory cytokines.•TLR2/TLR4 recognize scorpion venoms from the Brazilian Amazon and their antagonism abolish cytokine production.•The axis TLR-MyD88 is evolved in macrophage activation induced by scorpion venoms from the Brazilian Amazon.
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