Glucocerebrosidase reduces the spread of protein aggregation in a Drosophila melanogaster model of neurodegeneration by regulating proteins trafficked by extracellular vesicles

Abnormal protein aggregation within neurons is a key pathologic feature of Parkinson’s disease (PD). The spread of brain protein aggregates is associated with clinical disease progression, but how this occurs remains unclear. Mutations in glucosidase, beta acid 1 (GBA), which encodes glucocerebrosidase (GCase), are the most penetrant common genetic risk factor for PD and dementia with Lewy bodies and associate with faster disease progression. To explore how GBA mutations influence pathogenesis, we previously created a Drosophila model of GBA deficiency (Gba1b) that manifests neurodegeneration and accelerated protein aggregation. Proteomic analysis of Gba1b mutants revealed dysregulation of proteins involved in extracellular vesicle (EV) biology, and we found altered protein composition of EVs from Gba1b mutants. Accordingly, we hypothesized that GBA may influence pathogenic protein aggregate spread via EVs. We found that accumulation of ubiquitinated proteins and Ref(2)P, Drosophila homologue of mammalian p62, were reduced in muscle and brain tissue of Gba1b flies by ectopic expression of wildtype GCase in muscle. Neuronal GCase expression also rescued protein aggregation both cell-autonomously in brain and non-cell-autonomously in muscle. Muscle-specific GBA expression reduced the elevated levels of EV-intrinsic proteins and Ref(2)P found in EVs from Gba1b flies. Perturbing EV biogenesis through neutral sphingomyelinase (nSMase), an enzyme important for EV release and ceramide metabolism, enhanced protein aggregation when knocked down in muscle, but did not modify Gba1b mutant protein aggregation when knocked down in neurons. Lipidomic analysis of nSMase knockdown on ceramide and glucosylceramide levels suggested that Gba1b mutant protein aggregation may depend on relative depletion of specific ceramide species often enriched in EVs. Finally, we identified ectopically expressed GCase within isolated EVs. Together, our findings suggest that GCase deficiency promotes accelerated protein aggregate spread between cells and tissues via dysregulated EVs, and EV-mediated trafficking of GCase may partially account for the reduction in aggregate spread.     

Characterization of aneuploidy in interspecific hybrid between Urochloa ruziziensis (R. Germ. & Evrard) Crins and Urochloa decumbens (Stapf) R. D. Webster

Molecular Biology Reports - The aim of the study was to characterize the type of aneuploidy present in the hybrid Urochloa ruziziensis × Urochloa decumbens and to confirm the...     

Visual signals or displacement activities? The function of visual displays in agonistic interactions in nocturnal tree frogs

The efficiency of intraspecific communication directly affects male reproductive success. Acoustic signaling is the primary form of communication in nocturnal anurans. However, visual signaling can also be important in social interactions. We tested the hypothesis that open environments favor visual signals in a territorial defense context, in a nocturnal tree frog. We established three treatments each with eight males of Hypsiboas albomarginatus: (1) Clear Vision, with a mirror without visual obstacles; (2) Obstructed Vision, with half the mirror covered, and (3) Control, with mirror completely covered. We classified behavioral responses into orientation/locomotion, visual display, or acoustic signal. We calculated the mean emission rate per minute per behavior in each treatment and compared them among treatments using one-way ANOVA. Orientation and locomotion, visual display, and the advertisement call did not differ among treatments. However, the emission of aggressive calls in the Obstructed Vision treatment was significantly higher than in the Clear Vision treatment. The lowest rate of aggressive calls occurred in the Control. Thus, visual recognition of an intruder male was enough for resident males to adjust their rate of emission of acoustic aggressive signals, but not visual displays. Therefore, the recognition of the intruder male is not the only feature required for the evolution of visual signals in nocturnal tree frogs during agonistic interactions. This suggests that some visual displays may not be directly used for communication but rather constitute displacement activity.     

Chromosome painting of Z and W sex chromosomes in Characidium (Characiformes, Crenuchidae)

Some species of the genus Characidium have heteromorphic ZZ/ZW sex chromosomes with a totally heterochromatic W chromosome. Methods for chromosome microdissection associated with chromosome painting have become important tools for cytogenetic studies in Neotropical fish. In Characidium cf. fasciatum, the Z chromosome contains a pericentromeric heterochromatin block, whereas the W chromosome is completely heterochromatic. Therefore, a probe was produced from the W chromosome through microdissection and degenerate oligonucleotide-primed polymerase chain reaction amplification. FISH was performed using the W probe on the chromosomes of specimens of this species. This revealed expressive marks in the pericentromeric region of the Z chromosome as well as a completely painted W chromosome. When applying the same probe on chromosome preparations of C. cf. gomesi and Characidium sp., a pattern similar to C. cf. fasciatum was found, while C. cf. zebra, C. cf. lagosantense and Crenuchus spilurus species showed no hybridization signals. Structural changes in the chromosomes of an ancestral sexual system in the group that includes the species C. cf. gomesi, C. cf. fasciatum and Characidium sp., could have contributed to the process of speciation and could represent a causal mechanism of chromosomal diversification in this group. The heterochromatinization process possibly began in homomorphic and homologous chromosomes of an ancestral form, and this process could have given rise to the current patterns found in the species with sex chromosome heteromorphism.     

BTEX removal in a horizontal-flow anaerobic immobilized biomass reactor under denitrifying conditions

Because benzene, toluene, ethylbenzene, and xylenes (BTEX) and ethanol are important contaminants present in Brazilian gasoline, it is essential to develop technology that can be used in the bioremediation of gasoline-contaminated aquifers. This paper evaluates the performance of a horizontal-flow anaerobic immobilized biomass (HAIB) reactor fed with water containing gasoline constituents under denitrifying conditions. Two HAIB reactors filled with polyurethane foam matrices (5 mm cubes, 23 kg/m³ density and 95 % porosity) for biomass attachment were assayed. The reactor fed with synthetic substrate containing protein, carbohydrates, sodium bicarbonate and BTEX solution in ethanol, at an Hydraulic retention time (HRT) of 13.5 h, presented hydrocarbon removal efficiencies of 99 % at the following initial concentrations: benzene 6.7 mg/L, toluene 4.9 mg/L, m-xylene and p-xylene 7.2 mg/L, ethylbenzene 3.7 mg/L, and nitrate 60 mg N/L. The HAIB reactor fed with gasoline-contaminated water at an HRT of 20 h showed hydrocarbon removal efficiencies of 96 % at the following initial concentrations: benzene, 4.9 mg/L; toluene, 7.2 mg/L; m-xylene, 3.7 mg/L; and nitrate 400 mg N/L. Microbiological observations along the length of the HAIB reactor fed with gasoline-contaminated water confirmed that in the first segment of the reactor, denitrifying metabolism predominated, whereas from the first sampling port on, the metabolism observed was predominantly methanogenic.     

Species delimitation in Neotropical fishes of the genus Characidium (Teleostei,Characiformes)

Neotropical darters of the genus Characidium have a complex systematic history with several examples of sympatry throughout their distribution range in Neotropical freshwaters. Although various species within the genus have been used as models to investigate chromosomal evolution and biogeography, species boundaries and relationships still remain uncertain. Here, we use mitochondrial DNA sequences to perform species delimitation analyses within Characidium and test previous hypotheses of species richness within the Characidium zebra complex and among sympatric morphotypes of C. alipioi. Results indicate high genetic distances within tested species complexes and revealed the presence of strongly supported lineages such as the large C. lauroi group from southeastern Brazil. This suggests that the evolutionary history of these groups may be correlated with biogeographic history. Analyses also reveal that three geographically isolated populations of C. zebra represent a single species, leading us to reject prior hypothesis of multiple species. Species delimitation using mitochondrial data strongly supports the presence of two sympatric species within C. alipioi in southeastern Brazil despite limited morphological variation and conserved chromosomal patterns. These results provide a framework to further the study of systematics and evolution within Characidium.     

Detection of ‘long-haired’ Saprolegnia (S. parasitica) isolates using monoclonal antibodies

The ability of five monoclonal antibodies (Mabs) raised against a pathogenic Saprolegnia parasitica isolate from brown trout to detect and differentiate between isolates with bundles of long hairs (S. parasitica) and other Saprolegnia species was determined by means of an indirect immunofluorescence assay. Four of the Mabs used recognized some of the long-haired S. parasitica isolates but also cross-reacted with other Saprolegnia species without bundles of hairs and with Achlya sp. The other Mab (named 18A6) was able to differentiate between the asexual and most of the sexual isolates in the group of long-haired S. parasitica isolates, but did not recognize Achlya sp. or the Saprolegnia species without bundles of hairs, with the exception of S. hypogyna. These results indicate that isolates with bundles of long hairs are closely related with other members of genus Saprolegnia and share several antigens. However, Mab 18A6 seems to recognize an epitope that is expressed mainly in the asexual isolates in the long-haired S. parasitica isolates.     

Establishment of rapid and non-invasive protocols to identify B-carrying individuals of Psalidodon paranae

Supernumerary, or B, chromosomes are present in several eukaryotes, including characid fish of the genus Psalidodon. Notably, Psalidodon paranae carries the most studied B chromosome variant, a macro-B chromosome. The origin of this element was determined to be an isochromosome; however, data regarding its inheritance remain unavailable due to methodological barriers such as the lack of an efficient, non-invasive, and rapid protocol for identifying B-carrying individuals that would enable the design of efficient crossing experiments. Thus, in this study, we primarily aimed was to develop two non-invasive and fast (approximately 2 h) methods to identify the presence of B chromosomes in live specimens of P. paranae based on satellite DNA (satDNA) sequences known to be present in this element. The methods include fluorescence in situ hybridization in interphase nuclei and relative gene quantification of satDNAs using quantitative polymerase chain reaction. Our results reveal the efficiency of quick-fluorescence in situ hybridization and quantitative polymerase chain reaction for identifying B-carrying individuals using the proposed satDNA sequences and open up new possibilities to study B chromosomes.     

Phytoremediation applications in natural condition and in mesocosm: The uptake of cadmium by Lemna minuta Kunth,a non-native species in Italian watercourses

Metal pollution in water and soil is an environmental and public health issue. Cadmium (Cd) is included in the list of priority hazardous substances in the European Water Framework Directive. Phytoremediation system is a cost-effective, plant-based approach that takes advantage of the ability of plants to concentrate elements and compounds from the environment and to metabolize various molecules in their tissues. We studied the presence and the importance of an invasive species, such as Lemna minuta, in the environment and the effects of Cd pollution on this species. Growth, removal, and tolerance were evaluated for different Cd concentrations and different times of plant exposure. Overall, the results show that L. minuta has a good capacity of growth, metal bioconcentration, and tolerance up to 3 days of exposure at 0.5 and 1.5 mg L^?1 of Cd. In particular, L. minuta was able to accumulate Cd up to 3771 mg kg^?1 on dry mass basis. We can conclude that L. minuta possesses a great capability of Cd absorption and accumulation, thus supporting a potential use of this species in designing a metal bioremediation system in phytoremediation field.     

Molecular cytogenetic analysis and the establishment of a cell culture in the fish species Hollandichthys multifasciatus (Eigenmann & Norris, 1900) (Characiformes,Characidae)

Hollandichthys is a fish genus of the family Characidae that was until recently considered to be monotypic, with cytogenetic, morphological, and molecular data being restricted to a few local populations. In the present study, the karyotype of a population of Hollandichthys multifasciatus was analyzed using classical and molecular cytogenetic approaches for the investigation of potential markers that could provide new perspectives on the cytotaxonomy. H. multifasciatus presented a diploid number of 2n=50 chromosomes and a karyotype formula of 8m+10sm+32st. A single pair of chromosomes presented Ag-NORs signals, which coincided with the 18S rDNA sites visualized by FISH, whilst the 5S rDNA sequences were mapped in two chromosome pairs. The distribution of the U snRNA genes was mapped on the Hollandichthys chromosomes for the first time, with the probes revealing the presence of the U1 snDNA on the chromosomes of pair 20, U2 on pairs 6 and 19, U4 on pair 16, and U6 on the chromosomes of pair 11. The results of the present study indicated karyotypic differences in comparison with the other populations of H. multifasciatus studied previously, reinforcing the need for further research to identify isolated populations or the potential existence of cryptic Hollandichthys species.