Orientador: Fabio Porto-Foresti / Resumo: Dentre os peixes Neotropicais, o gênero Astyanax é caracterizado como um dos mais comuns e diversificados, representado por 147 espécies, das quais pelo menos 12 destas espécies têm demonstrado a presença de elementos adicionais denominados cromossomos B em seu genoma. No entanto, muitas vezes estas informações encontram-se dispersas ou fragmentadas. Assim, no presente estudo reportamos a diversidade de cromossomos B no gênero Astyanax, projetando nichos ecológicos preferenciais entre as espécies e populações B+ e B- do grupo em cenários do passado (Last Glacial Maximum - LGM: 22.000 e Mid-Holoceno - MD: 6.000 anos atrás), presente e futuro (2.060), além de análises populacionais de A. bockmanni a partir do mtDNA D-loop. Os períodos LGM e MH registraram ampla ocorrência de populações B+ e B- em boa parte das regiões Sudeste, Norte, Centro-Oeste e parte da Bahia. No presente observamos áreas favoráveis B+ e B- quase que exclusivamente em parte de SP, MG, PR, RJ, MT e ES. Para 2.060, verificamos áreas preferenciais B+ quase que exclusivamente em parte de MG, RJ e SP. Quando sobrepostos, os cenários indicam um acúmulo das populações B+ em uma mesma área, padrão não observado nas populações B-, sugerindo que a região sobreposta poderia representar uma zona climaticamente estável para populações B+ desde o LGM, já que nenhuma população B- foi mantida nesta região. Análises qPCR indicaram a presença de cromossomos BM em seis populações de A. bockmanni. A partir do mtDNA D-loop em 1... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Among the Neotropical fish, the genus Astyanax is characterized as the most common and diversified, represented by 147 species, of which at least 12 of these species have demonstrated the presence of additional elements denominated B chromosomes in their genome. However, this information is often scattered or fragmented. Thus, in the present study we report the diversity of B chromosomes in the Astyanax genus, projecting preferential ecological niches among the species and B+ and B- populations of the group in scenarios of the past (Last Glacial Maximum - LGM: 22.000 and MidHoloceno - MD: 6.000 years ago), present and e future (2.060), in addition to population analyzes of A. bockmanni from the mtDNA D-loop. The LGM and MH periods recorded a large occurrence of B+ and B- populations in most of the Southeast, North, Central West regions and part of Bahia State. In the present we observed favorable areas B+ and B- almost exclusively in part of SP, MG, PR, RJ, MT and ES. For 2.060, we see B+ preferred areas almost exclusively in part of MG, RJ and SP States. When overlapping, the scenarios indicate an accumulation of B+ populations in the same area, a pattern not observed in B- populations, suggesting that the overlapping region could represent a climatically stable zone for B+ populations since the LGM period, since no B- population was maintained in this region. qPCR analyzes indicated the presence of BM chromosomes in six A. bockmanni populations. From the D-loop mtDNA in 16 ... (Complete abstract click electronic access below) / Doutor

The nurse shark, Ginglymostoma cirratum, is a sedentary shark species that inhabits coral reefs in the tropical and subtropical Atlantic Ocean and along the western coast of the Americas in the Pacific Ocean. Nurse shark tissue samples were collected from the Bahamas, Belize, Brazil, and Dry Tortugas National Park in Florida. 186 individuals were genotyped at 11 microsatellite loci, the control region of the mitochondrial genome was sequenced in 190 individuals, and 89 individuals from the Bahamas, Belize, and Dry Tortugas were genotyped at the major histocompatibility complex (MHC) class IIα locus. An analysis of molecular variance (AMOVA) for the microsatellite loci indicated significant subdivision only between the Bahamas and Dry Tortugas populations. An AMOVA for the mitochondrial control region sequences indicated significant subdivision between all population pairs. The AMOVA for MHC class IIα locus indicated significant subdivision between two population pairs: the Bahamas population and the Dry Tortugas population and the Belize population and the Dry Tortugas population. The nurse shark has the lowest mitochondrial DNA nucleotide diversity (π=0.0125%) and haplotype diversity (h=0.2402) of any shark species to date. There were 14 MHC alleles from 39 polymorphic sites; ten were the same as published alleles (Kasahara et al. 1993; Ohta et al. 2000). This study was the first study to use MHC class IIα genes as a marker for population genetics in sharks. Our results showed that MHC class IIα locus behaves as a diploid locus and is a powerful tool for determining population genetic structure between populations.

O gênero Partamona compreende 33 espécies, distribuídas do sul do México ao sul do Brasil. O gênero tem sido amplamente estudado em diferentes níveis: citogenético, etológico e morfológico. O presente trabalho teve como objetivo contribuir com dados moleculares para o conhecimento do grupo, realizando estudos populacionais por meio da caracterização do DNA mitocondrial por PCR+RFLP e da análise de regiões de microssatélites do DNA genômico de duas espécies: P. mulata de distribuição restrita ao sul de Mato Grosso e norte do Mato Grosso do Sul, e P. helleri de distribuição mais ampla, do sul da Bahia até Santa Catarina. Foram detectados apenas dois haplótipos em P. mulata, os quais diferiram entre si por apenas um sítio de restrição. As análises estatísticas demonstraram não haver estruturação entre as populações sugerindo que esta espécie possa ter passado por recente afunilamento populacional. Em P. helleri foram observados dez haplótipos sendo alguns exclusivos e outros compartilhados. Análises estatísticas apontaram alta estruturação entre as populações e a distribuição filogeográfica observada sugere um possível isolamento por fragmentação da Mata Atlântica durante o Pleistoceno. A análise dos locos microssatélites mostrou baixa variabilidade genética em ambas espécies e discreta estruturação entre as populações, não relacionada com a distribuição geográfica das mesmas. Isto pode ser conseqüência de migração de machos entre populações visto que as rainhas são filopátricas ou, fragmentação dos habitats pela rápida degradação do cerrado e da Mata Atlântica, ou por alelos nulos causados pelo uso de primers heteroespecíficos. A análise de parentesco entre abelhas de um mesmo ninho apontou a existência de apenas uma patrilínea em P. mulata sugerindo monoandria para esta espécie. Foram encontradas duas patrilíneas em algumas colônias de P. helleri, o que pode ser resultante de fecundação por mais de um macho ou substituição recente da rainha. A caracterização parcial do DNAmt de duas espécies de Partamona poderá contribuir em estudos filogenéticos tanto do gênero quanto de outras espécies de Meliponini. A análise populacional mostrou o status da variabilidade genética das espécies, suas possíveis histórias evolutivas e a possível relação desta com degradação dos ambientes onde estas estão distribuídas. / The Partamona genus comprises 33 species distributed from south Mexico to south Brazil. This genus has been studied at different levels: cytogenetical, ethological and morphological. This work aimed at to contribute with molecular data for the knowledge about the group performing a population study employing the PCR+RFLP of mtDNA, and analysis of microsatellite loci from nuclear DNA of two species, P. mulata which is distributed in south Mato Grosso and north Mato Grosso do Sul, and P. helleri which geographic distribution is wider, from Santa Catarina to southern Bahia. It was detected two haplotypes in 58 colonies of P. mulata, each one differing by one single restriction site. The statistical analyses indicated no differentiation among populations suggesting that the species could have passed through a recent populational bottleneck. It was observed ten haplotypes in 47 colonies of P. helleri, some exclusive and others shared among populations. Statistical analysis pointed high population differentiation and the observed phylogeography distribution suggested a possible recent isolation probably by Atlantic Forest fragmentation during the Pleistocene. The microsatellite analysis showed low genetic variability in both species and discrete population structuring, not related to the geographic distribution. This might be consequence of migration of males, since the queens are highly phylopatric, or habitat fragmentation by degradation of savanna and Atlantic forest areas, or null alleles caused by the use of heterospecific primers. The relatedness investigation revealed only one patriline in nest mates of P. mulata that suggests monoandry for this species. It was found two patrilines in P. helleri that can be resulted from more than one mating or recent queen replacement. The partial characterization of the mtDNA of two Partamona species can contribute to further phylogenetic studies among bees of this genus or among other Meliponini species. The populational analysis showed the genetic variability status of the species, their putative evolutionary histories and the possible relation between the results and the environmental degradation in their distribution areas.

The primary objective of this proposal was to determine whether mitochondrial oxidative stress and variation in a particular mtDNA lineage contribute to the risk of developing cortical dysplasia and are potential contributing factors in epileptogenesis in children. The occurrence of epilepsy in children is highly associated with malformations of cortical development (MCD). It appears that MCD might arise from developmental errors due to environmental exposures in combination with inherited variation in response to environmental exposures and mitochondrial function. Therefore, it is postulated that variation in a particular mtDNA lineage of children contributes to the effects of mitochondrial DNA damage on MCD phenotype. Quantitative PCR and dot blot were used to examine mitochondrial oxidative damage and single nucleotide polymorphism (SNP) in the mitochondrial genome in brain tissue from 48 pediatric intractable epilepsy patients from Miami Children’s Hospital and 11 control samples from NICHD Brain and Tissue Bank for Developmental Disorders. Epilepsy patients showed higher mtDNA copy number compared to normal health subjects (controls). Oxidative mtDNA damage was lower in non-neoplastic but higher in neoplastic epilepsy patients compared to controls. There was a trend of lower mtDNA oxidative damage in the non-neoplastic (MCD) patients compared to controls, yet, the reverse was observed in neoplastic (MCD and Non-MCD) epilepsy patients. The presence of mtDNA SNP and haplogroups did not show any statistically significant relationships with epilepsy phenotypes. However, SNPs G9804A and G9952A were found in higher frequencies in epilepsy samples. Logistic regression analysis showed no relationship between mtDNA oxidative stress, mtDNA copy number, mitochondrial haplogroups and SNP variations with epilepsy in pediatric patients. The levels of mtDNA copy number and oxidative mtDNA damage and the SNPs G9952A and T10010C predicted neoplastic epilepsy, however, this was not significant due to a small sample size of pediatric subjects. Findings of this study indicate that an increase in mtDNA content may be compensatory mechanisms for defective mitochondria in intractable epilepsy and brain tumor. Further validation of these findings related to mitochondrial genotypes and mitochondrial dysfunction in pediatric epilepsy and MCD may lay the ground for the development of new therapies and prevention strategies during embryogenesis.

OPA1 regulates cristae structure and mitochondrial DNA (mtDNA) maintenance. Recently, our lab identified ATAD3A and SLC25 proteins as OPA1 interactors. After validating these interactions by co-immunoprecipitation, the role of these proteins in OPA1 function was examined. Previously, ATAD3A was implicated in mtDNA maintenance. However, no change in mtDNA content or nucleoid number was observed in my studies following long-term and short-term ATAD3A knockdown suggesting that OPA1 maintains mtDNA independently of ATAD3A. Previous data from our lab demonstrates that OPA1 oligomerization and cristae structure is altered by nutrients. SLC25 proteins transport nutrients into mitochondria. Therefore, OPA1 oligomerization and cristae structure was analyzed following SLC25 protein inhibition and knockdown. Decreased OPA1 oligomerization and cristae remodeling was observed following SLC25 protein inhibition and OGC knockdown. In addition these changes correlate with decreased ATP synthase monomers and oligomers suggesting that cristae remodeling may affect metabolism. Overall, these studies enhance our understanding of OPA1 function.

The Himalayan Mountain range encompasses an unparalleled landscape featuring some of the planet’s highest peaks, including Mount Everest. In the heart of this massive orographic barrier lies Nepal, sandwiched in the historically geostrategic position between the Tibetan plateau to the north and India in the south. Until recently, Nepalese and Tibetan populations remained poorly characterized genetically, partly because of their inaccessible geographical locations. In the present study, the genetic diversity of these two Himalayan populations is evaluated using different marker systems, including mitochondrial DNA (mtDNA) and Short Tandem Repeats (STRs) in the autosomes as well as on the Y-chromosome (Y-STR). While autosomal STRs are distributed throughout the genome and are biparentally inherited, the Y-chromosome and mtDNA are haploid markers and provide the paternal and maternal histories of the population, respectively. Fifteen autosomal STR loci were typed in 341 unrelated individuals from three Nepalese populations (188), namely Tamang (45), Newar (66) and Kathmandu (77), and a general collection from Tibet (153). These samples were also sequenced for the mtDNA control region and all of them were subsequently assigned to 75 different mtDNA haplogroups and sub-haplogroups by screening their diagnostic sites in the coding region using Restriction Fragment Length Polymorphism analysis and/or sequencing, thus achieving an unprecedented level of resolution. The results from the autosomal and mtDNA data suggest a Northeast Asian origin for the Himalayan populations, with significant genetic influence from the Indian subcontinent in Kathmandu and Newar, corroborating our previous Y-chromosome study. In contrast, Tibet displays a limited Indian component, suggesting that the Himalayan massif acted as a natural barrier for gene flow from the south. The presence of ancient Indian mtDNA lineages in Nepal implies that the region may have been inhabited by the earliest settlers who initially populated South Asia. In addition, seventeen Y-STR loci were analyzed in 350 Tibetan males from three culturally defined regions of historical Tibet: Amdo (88), Kham (109) and U-Tsang (153). The results demonstrate that the 17 Y-STR loci studied are highly polymorphic in all the three Tibetan populations examined and hence are useful for forensic cases, paternity testing and population genetic studies.

According to the WHO, 38 million individuals were living with human immunodeficiency virus (HIV), 25.4 million of which were using antiretroviral therapy (ART) at the end of 2019. Despite ART-mediated suppression of viral replication, ART is not a cure and is associated with viral persistence, residual inflammation, and metabolic disturbances. Indeed, due to the presence of viral reservoirs, lifelong ART therapy is required to control viremia and prevent disease progression into acquired immune deficiency syndrome (AIDS). Successful ART treatment allows people living with HIV (PLHIV) to achieve a similar life expectancy to uninfected individuals. However, recent studies have illustrated the presence of increased comorbidities, such as accelerated, premature immune aging, in ART-controlled PLHIV compared to uninfected individuals. Studies suggest that both HIV-infection and ART-treatment lead to mitochondrial dysfunction, ultimately resulting in cellular exhaustion, senescence, and apoptosis. Since mitochondria are essential cellular organelles for energy homeostasis and cellular metabolism, their compromise leads to decreased oxidative phosphorylation (OXPHOS), ATP synthesis, gluconeogenesis, and beta-oxidation, abnormal cell homeostasis, increased oxidative stress, depolarization of the mitochondrial membrane potential, and upregulation of mitochondrial DNA mutations and cellular apoptosis. The progressive mitochondrial damage induced by HIV-infection and ART-treatment likely contributes to accelerated aging, senescence, and cellular dysfunction in PLHIV. This review discusses the connections between mitochondrial compromise and cellular dysfunction associated with HIV-and ART-induced toxicities, providing new insights into how HIV and current ART directly impact mitochondrial functions and contribute to cellular senescence and aging in PLHIV. Identifying this nexus and potential mechanisms may be beneficial in developing improved therapeutics for treating PLHIV.

No description available.

>Magister Scientiae - MSc / Bladder grasshoppers belong to the order Orthoptera, ancient family Pneumoridae and Superfamily Pneumoroidea. This small group of grasshoppers are sound producing, nocturnal, herbivorous grasshoppers endemic to the coastal regions of southern Africa. Very little genetic work has been done on these grasshoppers, and there is some taxonomic confusion regarding the validity of some species descriptions. The aim of this study was to provide much needed clarity on the true taxonomic diversity and polymorphic attributes within the Pneumoridae, focusing on selected taxa of uncertain status. Bladder grasshoppers show distinct discontinuous polymorphism, resulting in two clearly different male morphs utilizing two different mating strategies. Primary males make use of acoustic communication for mate location. Secondary males (alternate males) are significantly smaller and employ a “sneaker” or satellite strategy where they exploit the calling between duetting couples to locate the females before the primary male. Three species of bladder grasshoppers have been described (Parabullacris vansoni, Paraphysemacris spinosus and Pneumoracris browni) that only have an alternate male morph. The validity of these species descriptions has come into question with the discovery of alternate male morphs in at least three other species (Bullacris discolor, B. membracioides and B. obliqua). Thus, the species described by Dirsh (1963) may simply be alternate males of existing species. However, to date there have been no studies looking at the genetics of alternate males, which would definitively establish whether they are conspecific with primary males.

No / Accumulation of large-scale mitochondrial DNA (mtDNA) deletions and chronic, subclinical inflammation are concomitant during skin aging, thus raising the question of a causal link. To approach this, we generated mice expressing a mutant mitochondrial helicase (K320E-TWINKLE) in the epidermis to accelerate the accumulation of mtDNA deletions in this skin compartment. Mice displayed low amounts of large-scale deletions and a dramatic depletion of mtDNA in the epidermis and showed macroscopic signs of severe skin inflammation. The mtDNA alterations led to an imbalanced stoichiometry of mitochondrial respiratory chain complexes, inducing a unique combination of cytokine expression, causing a severe inflammatory phenotype, with massive immune cell infiltrates already before birth. Altogether, these data unraveled a previously unknown link between an imbalanced stoichiometry of the mitochondrial respiratory chain complexes and skin inflammation and suggest that severe respiratory chain dysfunction, as observed in few cells leading to a mosaic in aged tissues, might be involved in the development of chronic subclinical inflammation. / Deutsche Forschungsgemeinschaft (Wi 889/6-3 to RJW, SFB 829 A14 to RJW, Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases–CECAD to RJW, BR2304/9-1 to BB, and SFB 829 A1, A5, and Z2 to CMN) and the Center of Molecular Medicine Cologne of the Medical Faculty (CMMC, to RJW)