RESUMO
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.
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.
RESUMO
Charadriiformes represent one of the largest orders of birds; members of this order are diverse in morphology, behavior and reproduction, making them an excellent model for studying evolution. It is accepted that the avian putative ancestral karyotype, with 2n = 80, remains conserved for about 100 million years. So far, only a few species of Charadriiformes have been studied using molecular cytogenetics. Here, we performed chromosome painting on metphase chromosomes of two species of Charadriidae, Charadrius collaris and Vanellus chilensis, with whole chromosome paint probes from Burhinus oedicnemus. Charadrius collaris has a diploid number of 76, with both sex chromosomes being submetacentric. In V. chilensi a diploid number of 78 was identified, and the Z chromosome is submetacentric. Chromosome painting suggests that chromosome conservation is a characteristic common to the family Charadriidae. The results allowed a comparative analysis between the three suborders of Charadriiformes and the order Gruiformes using chromosome rearrangements to understand phylogenetic relationships between species and karyotypic evolution. However, the comparative analysis between the Charadriiformes suborders so far has not revealed any shared rearrangements, indicating that each suborder follows an independent evolutionary path, as previously proposed. Likewise, although the orders Charadriiformes and Gruiformes are placed on sister branches, they do not share any signature chromosomal rearrangements.
Assuntos
Anfípodes , Charadriiformes , Anfípodes/genética , Animais , Aves/genética , Charadriiformes/genética , Coloração Cromossômica/métodos , Evolução Molecular , Filogenia , Cromossomos Sexuais/genéticaRESUMO
BACKGROUND: The Scolopacidae family (Suborder Scolopaci, Charadriiformes) is composed of sandpipers and snipes; these birds are long-distance migrants that show great diversity in their behavior and habitat use. Cytogenetic studies in the Scolopacidae family show the highest diploid numbers for order Charadriiformes. This work analyzes for the first time the karyotype of Actitis macularius by classic cytogenetics and chromosome painting. RESULTS: The species has a diploid number of 92, composed mostly of telocentric pairs. This high 2n is greater than the proposed 80 for the avian ancestral putative karyotype (a common feature among Scolopaci), suggesting that fission rearrangements have formed smaller macrochromosomes and microchromosomes. Fluorescence in situ hybridization using Burhinus oedicnemus whole chromosome probes confirmed the fissions in pairs 1, 2, 3, 4 and 6 of macrochromosomes. CONCLUSION: Comparative analysis with other species of Charadriiformes studied by chromosome painting together with the molecular phylogenies for the order allowed us to raise hypotheses about the chromosomal evolution in suborder Scolopaci. From this, we can establish a clear idea of how chromosomal evolution occurred in this suborder.
Assuntos
Charadriiformes , Coloração Cromossômica , Animais , Aves/genética , Charadriiformes/genética , Evolução Molecular , Hibridização in Situ FluorescenteRESUMO
Semipalmated sandpiper (Calidris pusilla) migration to the Southern Hemisphere includes a 5-day non-stop flight over the Atlantic Ocean, whereas semipalmated plover (Charadrius semipalmatus) migration, to the same area, is largely over land, with stopovers for feeding and rest. We compared the number and 3D morphology of hippocampal astrocytes of Ch. semipalmatus before and after autumnal migration with those of C. pusilla to test the hypothesis that the contrasting migratory flights of these species could differentially shape hippocampal astrocyte number and morphology. We captured individuals from both species in the Bay of Fundy (Canada) and in the coastal region of Bragança (Brazil) and processed their brains for selective GFAP immunolabeling of astrocytes. Hierarchical cluster analysis of astrocyte morphological features distinguished two families of morphological phenotypes, named type I and type II, which were differentially affected after migratory flights. Stereological counts of hippocampal astrocytes demonstrated that the number of astrocytes decreased significantly in C. pusilla, but did not change in Ch. semipalmatus. In addition, C. pusilla and Ch. semipalmatus hippocampal astrocyte morphological features were differentially affected after autumnal migration. We evaluated whether astrocyte morphometric variables were influenced by phylogenetic differences between C. pusilla and Ch. semipalmatus, using phylogenetically independent contrast approach, and phylogenetic trees generated by nuclear and mitochondrial markers. Our findings suggest that phylogenetic differences do not explain the results and that contrasting long-distance migratory flights shape plasticity of type I and type II astrocytes in different ways, which may imply distinct physiological roles for these cells.
Assuntos
Astrócitos , Charadriiformes , Animais , Canadá , Hipocampo , Humanos , FilogeniaRESUMO
Astrocytes are essential for lipid neuronal metabolism in long-distance uninterrupted migratory flights, when glucose is not available as the main source of energy. We previously demonstrated in Calidris pusilla that after uninterrupted 5 days transatlantic flight, astrocytes shrink and reduce its number in the hippocampal formation. Here we shifted our attention to the wintering period and tested the hypothesis that hippocampal astrocyte morphology of A interpres will change as the wintering period progresses towards the premigration window. To that end we used Arenaria interpres, which also crosses the Atlantic Ocean and reaches the mangroves of the Amazon River estuary for wintering. Birds were captured in September/October (closer to the arrival in the coast of Bragança, Para, Brazil for wintering) and in April/May (closer to the departure towards the breeding sites) and had their brains processed for selective GFAP-astrocyte immunolabeling. Three-dimensional reconstructions of the immunostained astrocytes were performed and morphological classification was done based on hierarchical cluster and discriminant analysis of multimodal morphometric features. We found two morphological phenotypes of astrocytes in the newcomers which differentially increased its morphological complexities as wintering period progresses towards the pre-migration window. Taken together, our findings demonstrate that the long-distance non-stop flight and wintering period differentially affected the two astrocytes morphotypes, suggesting distinct physiological roles for these cells. We suggest that morphological changes during the wintering period, may be part of the adaptive plasticity of the local hippocampal circuits of A. interpres in preparation for the long journey back to their breeding sites in the north hemisphere.