Adult Viability Research Articles

Effects of temperature on the development of Rachiplusia nu (Lepidoptera: Noctuidae) and Chrysodeixis includens (Lepidoptera: Noctuidae) and implications on population growth in Brazil.

Temperature is an elementary component in mathematical models for predicting the biotic potential of insects. In this study, the objective was to evaluate the impact of different constant temperatures of 8, 10, 15, 20, 25, 30, and 32°C on the biological parameters, lower temperature thresholds (TT), and estimating the number of annual generations (NAG) of Rachiplusia nu and Chrysodeixis includens, both pests associated with the soybean crop in Brazil. There was no development of the immature stages of R. nu at 8°C, as was also found for C. includens at 8 and 10°C. However, at 10°C all stages of R. nu developed. In general, temperatures of 20 and 25°C were the most suitable for the development of R. nu and C. includens, providing egg to adult viability of over 60% and the highest total fecundities. However, the temperature of 32°C negatively affected the parameters of the fertility life table. Rachiplusia nu showed the lowest TT (eggs: 4.9°C; larvae: 10.8°C; pupa: 14.1°C; and egg to adult: 8.9°C) when compared with C. includens (eggs: 7.5°C; larvae: 15.3°C; pupa: 16.1°C; and egg to adult: 11.3°C). Based on TT values, the NAG varied from 3.9 in cold regions to 7.5 in warm regions. However, for C. includens, we can infer that the species can reach up to 8.8 generations in warm regions. The results of the present study are important for understanding the occurrence of R. nu and C. includens in field conditions and can help with the implications of management strategies.

Plant-Based Insecticides for Control of Aedes aegypti, Sitophilus zeamais and Nasutitermes corniger: A Review and Bibliometric Analysis

Insect pests can transmit diseases or affect the economy and their uncontrolled growth is intensified by human activity, which has affected ecosystems and interfered with trophic chains. Aedes aegypti L. transmits dengue, chikungunya, zika and yellow fever, while Sitophilus zeamais Motsch is a stored grain pest, and Nasutitermes corniger Motsch attacks trees and urban buildings. This work provides a review of the state of art and current trends, as well as a bibliometric analysis on the insecticidal activity of plant-based products against Ae. aegypti, S. zeamais and N. corniger. The review of literature data showed that plant insecticides comprise secondary metabolites and proteins that can impair the viability of embryos, larvae, pupae, and adults, as well as fertility and oviposition or feeding behavior of insects. Also, they tend to be more biodegradable and less harmful to non-target organisms compared to synthetic insecticides. The bibliometric analysis revealed the concentration of publications in tropical regions worldwide, where there is a strong occurrence of diseases transmitted by Ae. aegypti. These reports were far more numerous than those on N. corniger and S. zeamais. Furthermore, the synthesis of metallic nanoparticles from plant products has strongly emerged as a current trend.

Toxicity evaluation of novel imidacloprid nanoribbons, using somatic mutation and fitness indexes in Drosophila melanogaster

ABSTRACT Nanoribbons of imidacloprid, a systemic and chloronicotinyl insecticide, were successfully synthesized by laser-induced fragmentation/exfoliation of imidacloprid powders suspended in water, with widths ranging from 160 to 470 nm, lengths in the micron scale, and thickness of a few atoms layers. The aim of the present study was to examine the effects of acute and chronic exposure to imidacloprid (IMC) bulk and compare its effects with synthesized imidacloprid nanoribbons (IMCNR) on larval and adult viability, developmental time, olfactory capacity, longevity, productivity, and genotoxicity in Drosophila melanogaster. Larvae or adults were exposed at 0.01, 0.02, or 0.03 ppm to IMC or IMCNR. Results demonstrated that IMCNR produced a significant reduction in viability and olfactory ability. IMC did not significantly alter viability and olfactory ability. Similarly, marked differences on longevity were detected between treatment with IMC and IMCNR where the lifespan of males treated with IMC was significantly higher than control while IMCNR produced a reduction. As for productivity, developmental time, and genotoxicity, no marked differences were found between both forms of IMC.

Genotoxicity responses of single and mixed exposure to heavy metals (cadmium, silver, and copper) as environmental pollutants in Drosophila melanogaster

Heavy metals are now persistently present in living things' environments, in addition to their potential toxicity. Therefore, the aim of this study was to utilize D. melanogaster to determine the biological effects induced by different heavy metals including cadmium chloride (CdCl2), copper (II) sulfate pentahydrate (CuSO 4.5 H2O), and silver nitrate (AgNO3). In vivo experiments were conducted utilizing three low and environmentally relevant concentrations from 0.01 to 0.5 mM under single and combined exposure scenarios on D. melanogaster larvae. The endpoints measured included viability, reactive oxygen species (ROS) generation and genotoxic effects using Comet assay and the wing-spot test. Results indicated that tested heavy metals were not toxic in the egg-to adult viability. However, combined exposure (CdCl2+AgNO3 and CdCl2+AgNO3+CuSO 4.5 H2O) resulted in significant genotoxic and unfavorable consequences, as well as antagonistic and/or synergistic effects on oxidative damage and genetic damage.

STUDY OF THE EFFICIENCY AND COMPARATIVE ANALYSIS OF THE APPLICATION OF TECHNIQUES FOR EVALUATING THE QUALITY OF PHYTOPHAGES AND ENTOMOPHAGES AT DIFFERENT STAGES OF ONTOGENESIS

The concept of quality control of insect crops is proposed, which is based on a targeted approach based on the goal and objectives of the implementation of a certain breeding program and is divided into two main directions: control of phytophage production, control of entomophage production. Quality control methods are based on the general biological rule of dependence between the intensity of insect taxis and the overall viability of biomaterial. The effectiveness of methods for assessing the quality of phytophages (on the example of the mother cultures of the grain moth Sitotroga cerealella, the wax moth Galleria mellonella) and entomophages (laboratory and natural populations of P. bioculatus.) of the Center for Mother Cultures of Insects Engineering and Technological Institute “Biotekhnica” of the National Academy of Sciences (ETI “Biotekhnica”) at different stages of ontogenesis was investigated using the indicator the intensity of taxis. As a result of research conducted on the culture of phytophagous, grain moth, a significant difference with the control was established in the indicators of viability of eggs, larvae, pupae and adults: 25; 6; 33 and 11%, respectively. Studies conducted with another phytophagous representative – the wax firefly – also showed positive results (16; 17; 16; 11%, respectively). The obtained (as a result of preliminary quality control) and selection culture had significantly higher viability indicators at all stages of ontogenesis. It was established that the intensity of chemotaxis of larvae of the 1st age of entomophages directly correlates with the indicator of viability of the insect culture. When selecting the starting material for successful culture, it is possible to select by chemotaxis of the larvae after rebirth from the egg. Thus, viability when selecting for Colorado beetle eggs significantly (p<0.01) exceeds this indicator in other selection options – Colorado beetle larvae and wax firefly larvae: 82; 59; 73% respectively. When working with an artificial population of P. bioculatus, on wax moth larvae, the highest intensity of taxis was noted in the variant with wax moth larvae compared to Colorado beetle eggs and Colorado beetle larvae – 37; 34; 29% respectively. This indicates a shift in the number of the population in the direction of individuals that successfully feed on unnatural food. Their viability was significantly higher than the viability in other variants of the study with an artificial population – 78; 71; 68% respectively. The proposed method of selection of starting material for cultivation of P. bioculatus allows differentiation of individuals for further effective reproduction, and the use of such biomaterial in programs of biological protection of plants from pests. A comparative analysis of methods for assessing the quality of phytophagous and entomophagous cultures at different stages of ontogenesis was carried out using the indicator of the overall viability of insect populations. It is an integrated indicator that takes into account the viability of the population at all stages of its development. All tested objects of the study showed that control and improvement of culture quality by the proposed method lead to an increase in the viability of biomaterial by 18.71; 18.56 and 8.86%, respectively (egg, larva, pupa).

Functional characterisation of the ACE2 orthologues in Drosophila provides insights into the neuromuscular complications of COVID-19

SARS-CoV-2, the virus responsible for the coronavirus disease of 2019 (COVID-19), gains cellular entry via interaction with the angiotensin-converting enzyme 2 (ACE2) receptor of host cells. Although SARS-CoV-2 mainly targets the respiratory system, the neuromuscular system also appears to be affected in a large percentage of patients with acute or chronic COVID-19. The cause of the well-described neuromuscular manifestations resulting from SARS-CoV-2 infection remains unresolved. These may result from the neuromuscular-invasive capacity of the virus leading to direct injury. Alternatively, they may be the consequence of ACE2 inactivation either due to viral infection, ACE2 autoantibodies or both. Here, we made use of the Drosophila model to investigate whether ACE2 downregulation is sufficient to induce neuromuscular phenotypes. We show that moderate gene silencing of ACE2 orthologues Ance or Ance3 diminished survival on exposure to thermal stress only upon induction of neuromuscular fatigue driven by increased physical activity. A strong knockdown of Ance or Ance3 directed to muscle reduced or abolished adult viability and caused obvious motoric deficits including reduced locomotion and impaired flight capacity. Selective knockdown of Ance and Ance3 in neurons caused wing defects and an age-dependent decline in motor behaviour, respectively, in adult flies. Interestingly, RNA sequencing allowed us to discover several differentially spliced genes that are required for synaptic function downstream of Ance or Ance3 depletion. Our findings are therefore supportive of the notion that loss of a RAS-independent function for ACE2 contributes to the neuromuscular manifestations associated with SARS-CoV-2 infection.

Selectivity of Bacillus thuringiensis (Bacillales: Bacillaceae) to the polyphagous predator Ceraeochrysa claveri (Navás, 1911) (Neuroptera: Chrysopidae)

AbstractBioinsecticides based on Bacillus thuringiensis (Bt) are widely used to control agricultural pests. However, there are concerns about their impact on non‐target organisms, with for example, the green lacewing Ceraeochrysa claveri, a polyphagous predator with important value in the biological control of insect pests in agroecosystems. This study aimed to analyse the effects of Agree® bioinsecticide (containing Bt serovar aizawai GC‐91) on the post‐embryonic development of C. claveri. For this, neonate larvae of C. claveri were fed with Diatraea saccharalis eggs contaminated with different concentrations of Agree® (0.62, 1.25, 2.50 and 5.00 g L−1) until their pupation. Samples of the midgut of larvae were collected for histological and ultrastructural analysis. The larval and pupal inviability data were collected at the end of the respective developmental stages. After the emergence of adult insects, were considered inviable those with morphological alterations. To evaluate the reproduction, adult insect couples were formed, and the oviposition was collected. No significant differences in the viability of eggs, larvae, pupae and adults of C. claveri contaminated with Agree® were reported. Midgut analyses did not reveal cellular damage in the epithelium, indicating a possible lack of insecticidal proteins receptors. The oviposition and their viability were similar between treated and control groups. Our results demonstrated that Agree® bioinsecticide did not affect the post‐embryonic development of C. claveri. This illustrates that the combined use of Bt and C. claveri for the biological control of arthropod pests can be a sustainable strategy for integrated pest management.

Obligate role for Rock1 and Rock2 in adult stem cell viability and function

The ability of stem cells to rapidly proliferate and differentiate is integral to the steady-state maintenance of tissues with high turnover such as the blood and intestine. Mutations that alter these processes can cause primary immunodeficiencies, malignancies and defects in barrier function. The Rho-kinases, Rock1 and Rock2, regulate cell shape and cytoskeletal rearrangement, activities essential to mitosis. Here, we use inducible gene targeting to ablate Rock1 and Rock2 in adult mice, and identify an obligate requirement for these enzymes in the preservation of the hematopoietic and gastrointestinal systems. Hematopoietic cell progenitors devoid of Rho-kinases display cell cycle arrest, blocking the differentiation to mature blood lineages. Similarly, these mice exhibit impaired epithelial cell renewal in the small intestine, which is ultimately fatal. Our data reveal a novel role for these kinases in the proliferation and viability of stem cells and their progenitors, which is vital to maintaining the steady-state integrity of these organ systems.

Effects of ELF-PEMF exposure on spontaneous alternation, anxiety, motor co-ordination and locomotor activity of adult wistar rats and viability of C6 (Glial) cells in culture

The effects of ELF-PEMF exposure on spontaneous alternation, anxiety, motor coordination, and locomotor activity have been discussed in various pre-clinical and clinical settings. Several epidemiological and experimental studies have demonstrated the potential effects of ELF-PEMF when exposed > ∼1 h/day; however, very few studies have focused on understanding the influence of ELF-PEMF exposure of 1–3 mT with an exposure duration of < 1 h/day on spontaneous alternation, anxiety, motor coordination, and locomotor activity. Hence, we attempted to study the effects of ELF-PEMF exposure of 1–3 mT, 50 Hz with an exposure duration of 20 min each with a 4 h gap (2 times) on the cellular proliferation and morphologies of C6 (Glial) cells and spontaneous alternation, anxiety, motor coordination and locomotor activity of Wistar rats under in vitro and in vivo conditions, respectively. The results showed that ELF-PEMF exposure did not induce any significant levels of cellular fragmentation and changes in the morphology of glial cells. Also, the outcomes revealed no noticeable effects on spontaneous alternation, anxiety, motor coordination, and locomotor activity in PEMF-exposed groups compared with the control. No undesirable side effects were observed at the highest dose (B=3 mT). We also performed histological analysis of the selected brain sections (hippocampus and cortex) following ELF-PEMF exposure. Incidentally, no significant changes were observed in cortical cell counts, tissue structure, and morphology.

Anti-Onchocercal Properties of Extracts of Scoparia dulcis and Cylicodiscus gabunensis.

The elimination of onchocerciasis is hampered by the absence of suitable drugs that are effective against adult filariae. This study is aimed at assessing the anti-onchocercal effects of extracts of Scoparia dulcis and Cylicodiscus gabunensis that could serve as drug leads against onchocerciasis. Different parts of the plants (Scoparia dulcis and Cylicodiscus gabunensis) were extracted with hexane, methylene chloride, and methanol. The extracts were tested in vitro against the bovine model parasite, Onchocerca ochengi. Adult female worm viability was determined biochemically by MTT/formazan colorimetry, while the adult male and microfilariae viability were determined by microscopy based on % inhibition of worm motility score. Cytotoxicity and acute toxicity of active extracts were tested on monkey kidney epithelial cells (LLC-MK2) and Balb/C mice, respectively. The hexane extract of Scoparia dulcis recorded the highest activity, with IC50s of 50.78 μg/ml on both adult male and female worms and 3.91 μg/ml on microfilariae. For Cylicodiscus gabunensis extract, the highest activity was seen with the methylene chloride extract, with IC50s of 50.78 μg/ml, 62.50 μg/ml, and 16.28 μg/ml on, respectively, adult male, female, and microfilariae. The 50% cytotoxic concentration on the LLC-MK2 cells was 31.25 μg/ml for the most active extracts. No acute toxicity was recorded for the extracts. Phytochemical analysis of the extracts revealed the presence of alkaloids, flavonoids, sterols, saponins, phenols, and glycosides. This study validates the traditional use of these plants in treating onchocerciasis and suggests S. dulcis and C. gabunensis as new potential sources for the isolation of anti-onchocerca lead compounds.

Developmental and reproduction toxicity of piperonyl butoxide part 3 two generation (two litter) reproduction study of piperonyl butoxide administered in the diet to CD ® (Sprague Dawley) rats

Piperonyl butoxide (PBO) an insecticide synergist was evaluated in a guideline multigenerational toxicity study in rats. F0 and F1 adult generations consisted of groups of 26 male and 26 female CD (Sprague Dawley) rats that were exposed to PBO in the diet at concentrations of 0, 300, 1000 or 5000 ppm for 85 (F0) or 83 (F1) days prior to cohabitation and throughout two mating periods (F1a, F2a and F1b, F2b). Exposure to test diets continued through the mating, gestation, and lactation periods for the females. F2 generation pups were euthanized following weaning. There were no effects on survival, clinical observations, gross or histological findings, fertility, pup viability, lactation indices or sex ratio in adults or pups in any generation. All effects of PBO occurred in the 5000-ppm exposure group. These effects included reduced body weight gains for F0 and F1 males and females during pre-cohabitation resulting in reduced body weights during both gestation periods. Food consumption of the F1b group males was slightly or significantly less than control values from week 3 onward. F1a generation pup weights were reduced on days 4, 7, 14 and 21 postpartum. Pup weights in the F1 and F2 generations were significantly reduced on days 14 and 21 postpartum when diets were being consumed by pups. The no-observable-adverse-effect level (NOAEL) for general toxicity was 1000 ppm based on reductions in body weights (parental and offspring) at 5000 ppm; and the NOAEL for reproductive toxicity was 5000 ppm with no direct effects on reproduction.

The WALLET Study: Examining Early Memory Loss and Personal Finance.

Background and ObjectivesThis feasibility study tests a new approach for assessing personal finance in older persons with early memory loss. The project examines 2 primary outcomes that gauge the financial viability and well-being of older adults: wealth loss and financial exploitation. The overall objective is to determine the association of financial literacy and management, financial decision-making, and cognition with wealth loss and financial exploitation.Research Design and MethodsThis cross-sectional study recruited 46 participants who were 60 years of age or older. Participants were classified as having mild cognitive impairment, perceived cognitive impairment, or no cognitive impairment. The study coordinator arranged with each participant to obtain copies of their main checking account statements for 12 consecutive months within the previous 2 years and, if appropriate, credit card statements. All statements were de-identified and assigned a random ID number. Participants then completed 2 telephone interviews.ResultsThe average participant age was 72 years (standard deviation [SD] = 7.7); 84% were female, 39% White, and 35% currently married. Average education was 16.2 years (SD = 2.4); mean yearly household income was almost $42,000 (SD = 25,752); and monthly social security payments averaged $1,446 (SD = 1,244). Our results indicate that the methods used to analyze checking account statements, followed by telephone interviews to verify identified trends, were useful in developing a financial behavior index to measure wealth loss.Discussion and ImplicationsWe demonstrate an alternative method for assessing personal finance using person-centered principles, which we believe are critical in the presence of diminished or impaired cognition. Our findings offer an innovative method for assessing the risk for wealth loss and financial exploitation.

RNAi in Piezodorus guildinii (Hemiptera: Pentatomidae): Transcriptome Assembly for the Development of Pest Control Strategies.

Red-banded stink bug Piezodorus guildinii (P. guildinii) has been described as the most damaging stink bug regarding soybean crops, leading to seed injury, low germination percentages, and foliar retention, at low population densities. In recent years, RNA interference (RNAi), a conserved eukaryote silencing mechanism has been explored to develop species-selective pesticides. In this work, we evaluated RNAi in P. guildinii to develop new pest-control strategies. For this, we assembled and annotated a P. guildinii transcriptome from a pool of all developmental stages. Analysis of this transcriptome led to the identification of 56 genes related to the silencing process encompassing siRNA, miRNA, and piRNA pathways. To evaluate the functionality of RNAi machinery, P. guildinii adults were injected with 28 ng/mg of body weight of double stranded RNA (dsRNA) targeting vATPase A. A mortality of 35 and 51.6% was observed after 7 and 14 days, respectively, and a downregulation of vATPase A gene of 84% 72 h post-injection. In addition, Dicer-2 and Argonaute-2 genes, core RNAi proteins, were upregulated 1.8-fold 48 h after injection. These findings showed for the first time that RNAi is functional in P. guildinii and the silencing of essential genes has a significant effect in adult viability. Taken together, the work reported here shows that RNAi could be an interesting approach for the development of red-banded stink bug control strategies.

Assessing the Anthelmintic Candidates BLK127 and HBK4 for Their Efficacy on Haemonchus contortus Adults and Eggs, and Their Hepatotoxicity and Biotransformation.

As a widely distributed parasitic nematode of ruminants, Haemonchus contortus has become resistant to most anthelmintic classes, there has been a major demand for new compounds against H. contortus and related nematodes. Recent phenotypic screening has revealed two compounds, designated as BLK127 and HBK4, that are active against H. contortus larvae. The present study was designed to assess the activity of these compounds against H. contortus eggs and adults, hepatotoxicity in rats and sheep, as well as biotransformation in H. contortus adults and the ovine liver. Both compounds exhibited no inhibitory effect on the hatching of eggs. The benzyloxy amide BLK127 significantly decreased the viability of adults in sensitive and resistant strains of H. contortus and showed no hepatotoxic effect, even at the highest concentration tested (100 µM). In contrast, HBK4 had no impact on the viability of H. contortus adults and exhibited significant hepatotoxicity. Based on these findings, HBK4 was excluded from further studies, while BLK127 seems to be a potential candidate for a new anthelmintic. Consequently, biotransformation of BLK127 was tested in H. contortus adults and the ovine liver. In H. contortus, several metabolites formed via hydroxylation, hydrolysis and glycosidation were identified, but the extent of biotransformation was low, and the total quantity of the metabolites formed did not differ significantly between the sensitive and resistant strains. In contrast, ovine liver cells metabolized BLK127 more extensively with a glycine conjugate of 4-(pentyloxy)benzoic acid as the main BLK127 metabolite.

Disruption of Survival Motor Neuron in Glia Impacts Survival but has no Effect on Neuromuscular Function in Drosophila

Increasing evidence points to the involvement of cell types other than motor neurons in both amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA), the predominant motor neuron disease in adults and infants, respectively. The contribution of glia to ALS pathophysiology is well documented. Studies have since focused on evaluating the contribution of glia in SMA. Here, we made use of the Drosophila model to ask whether the survival motor neuron (Smn) protein, the causative factor for SMA, is required selectively in glia. We show that the specific loss of Smn function in glia during development reduced survival to adulthood but did not affect motoric performance or neuromuscular junction (NMJ) morphology in flies. In contrast, gain rather than loss of ALS-linked TDP-43, FUS or C9orf72 function in glia induced significant defects in motor behaviour in addition to reduced survival. Furthermore, glia-specific gain of TDP-43 function caused both NMJ defects and muscle atrophy. Smn together with Gemins 2–8 and Unrip, form the Smn complex which is indispensable for the assembly of spliceosomal small nuclear ribonucleoproteins (snRNPs). We show that glial-selective perturbation of Smn complex components or disruption of key snRNP biogenesis factors pICln and Tgs1, induce deleterious effects on adult fly viability but, similar to Smn reduction, had no negative effect on neuromuscular function. Our findings suggest that the role of Smn in snRNP biogenesis as part of the Smn complex is required in glia for the survival of the organism, underscoring the importance of glial cells in SMA disease formation.

Sodium chloride reduces the viability of eggs, larvae, and pupae of Spodoptera frugiperda, as well as delays its development

The control of fall armyworm (Spodoptera frugiperda) populations involves the application of pesticides that cause several unwanted effects to humans and other animals, as well as to environment. Sodium chloride (NaCl) has been used in agriculture to potentiate the toxicity of pesticides. Here, the effects of NaCl on the viability of eggs, larvae, and pupae, as well as on development of S. frugiperda were evaluated. The treatment with 0.15 M NaCl caused a 35% reduction of egg hatching and the ingestion of artificial diet containing the salt caused mortality of caterpillars, pre-pupae, and pupae, as well as delayed the development of the insects. The larval, pre-pupa and pupa stages were elongated by about 2, 1 and 0.8 days, respectively. Additionally, significant reduction in pupae body weight was recorded in groups treated with NaCl at 0.62 mg/g and 3.12 mg/g. Despite these effects, NaCl ingestion did not affect the viability of adults and their reproductive performance. Food deterrent assay revealed that the salt acted as a phagostimulant agent. In conclusion, NaCl reduces the hatching of S. frugiperda eggs and damage the survival of carterpillars and pupae, as well interferes with insect development.

The Putative Drosophila TMEM184B Ortholog Tmep Ensures Proper Locomotion by Restraining Ectopic Firing at the Neuromuscular Junction.

TMEM184B is a putative seven-pass membrane protein that promotes axon degeneration after injury. TMEM184B mutation causes aberrant neuromuscular architecture and sensory and motor behavioral defects in mice. The mechanism through which TMEM184B causes neuromuscular defects is unknown. We employed Drosophila melanogaster to investigate the function of the closely related gene, Tmep (CG12004), at the neuromuscular junction. We show that Tmep is required for full adult viability and efficient larval locomotion. Tmep mutant larvae have a reduced body contraction rate compared to controls, with stronger deficits in females. In recordings from body wall muscles, Tmep mutants show substantial hyperexcitability, with many postsynaptic potentials fired in response to a single stimulation, consistent with a role for Tmep in restraining synaptic excitability. Additional branches and satellite boutons at Tmep mutant neuromuscular junctions are consistent with an activity-dependent synaptic overgrowth. Tmep is expressed in endosomes and synaptic vesicles within motor neurons, suggesting a possible role in synaptic membrane trafficking. Using RNAi knockdown, we show that Tmep is required in motor neurons for proper larval locomotion and excitability, and that its reduction increases levels of presynaptic calcium. Locomotor defects can be rescued by presynaptic knockdown of endoplasmic reticulum calcium channels or by reducing evoked release probability, further suggesting that excess synaptic activity drives behavioral deficiencies. Our work establishes a critical function for Tmep in the regulation of synaptic transmission and locomotor behavior.

Two Isoforms of serpent Containing Either One or Two GATA Zinc Fingers Provide Functional Diversity During Drosophila Development.

GATA transcription factors play crucial roles in various developmental processes in organisms ranging from flies to humans. In mammals, GATA factors are characterized by the presence of two highly conserved domains, the N-terminal (N-ZnF) and the C-terminal (C-ZnF) zinc fingers. The Drosophila GATA factor Serpent (Srp) is produced in different isoforms that contains either both N-ZnF and C-ZnF (SrpNC) or only the C-ZnF (SrpC). Here, we investigated the functional roles ensured by each of these isoforms during Drosophila development. Using the CRISPR/Cas9 technique, we generated new mutant fly lines deleted for one (ΔsrpNC) or the other (ΔsrpC) encoded isoform, and a third one with a single point mutation in the N-ZnF that alters its interaction with its cofactor, the Drosophila FOG homolog U-shaped (Ush). Analysis of these mutants revealed that the Srp zinc fingers are differentially required for Srp to fulfill its functions. While SrpC is essential for embryo to adult viability, SrpNC, which is the closest conserved isoform to that of vertebrates, is not. However, to ensure its specific functions in larval hematopoiesis and fertility, Srp requires the presence of both N- and C-ZnF (SrpNC) and interaction with its cofactor Ush. Our results also reveal that in vivo the presence of N-ZnF restricts rather than extends the ability of GATA factors to regulate the repertoire of C-ZnF bound target genes.

The disease-associated proteins Drosophila Nab2 and Ataxin-2 interact with shared RNAs and coregulate neuronal morphology.

Nab2 encodes the Drosophila melanogaster member of a conserved family of zinc finger polyadenosine RNA-binding proteins (RBPs) linked to multiple steps in post-transcriptional regulation. Mutation of the Nab2 human ortholog ZC3H14 gives rise to an autosomal recessive intellectual disability but understanding of Nab2/ZC3H14 function in metazoan nervous systems is limited, in part because no comprehensive identification of metazoan Nab2/ZC3H14-associated RNA transcripts has yet been conducted. Moreover, many Nab2/ZC3H14 functional protein partnerships remain unidentified. Here, we present evidence that Nab2 genetically interacts with Ataxin-2 (Atx2), which encodes a neuronal translational regulator, and that these factors coordinately regulate neuronal morphology, circadian behavior, and adult viability. We then present the first high-throughput identifications of Nab2- and Atx2-associated RNAs in Drosophila brain neurons using RNA immunoprecipitation-sequencing (RIP-Seq). Critically, the RNA interactomes of each RBP overlap, and Nab2 exhibits high specificity in its RNA associations in neurons in vivo, associating with a small fraction of all polyadenylated RNAs. The identities of shared associated transcripts (e.g., drk, me31B, stai) and of transcripts specific to Nab2 or Atx2 (e.g., Arpc2 and tea) promise insight into neuronal functions of, and genetic interactions between, each RBP. Consistent with prior biochemical studies, Nab2-associated neuronal RNAs are overrepresented for internal A-rich motifs, suggesting these sequences may partially mediate Nab2 target selection. These data support a model where Nab2 functionally opposes Atx2 in neurons, demonstrate Nab2 shares associated neuronal RNAs with Atx2, and reveal Drosophila Nab2 associates with a more specific subset of polyadenylated mRNAs than its polyadenosine affinity alone may suggest.

Cell death induction of dibutyl phthalate (DBP) on primary brain cells from adult zebrafish

The presence of phthalates as plasticizer on pharmaceutical products as well as in aquatic environments requires serious attention. Phthalates are compounds widely used in the manufacturing industry and has been associate with multiple adverse health effects to human. Among the family of phthalates Dibutyl phthalate (DBP) is well used for pharmaceutical products. Limited information however has been generated on the effect of DBP on the brain. The objective of this investigation was to characterize potential neurotoxicity resulting from short term exposure to DBP. Considering the efficacy and sensitivity of the zebrafish model as an early stage- screening tool, and their neurological and biochemical similarities to those of human, this study evaluates the effect of DBP on primary brain cells of the adult zebrafish. This well optimized and implemented methodology can be consider a guide for future studies using primary cells of adult zebrafish given the reduction on reagents, and the number of cells used for treatment. For indirect confirmation of extracted nervous tissue, the Acetylcholinesterase activity (AChEs) assay kit was implemented. In our study primary cells from adult zebrafish brain were exposed to DBP for 24 hours at aerobic environment at concentrations from 3uM to 100μM. The effect on cell viability as well as cell death mechanisms was determined. Results indicate that adult primary brain cells viability is compromised as low as 8uM of DBP. In terms of mechanism, cells undergo apoptosis and autophagy at concentrations of 8μM and 100μM. This study confirms that low concentrations of DBP over a 24-hour period induces apoptosis and autophagy, potentially activating degradation of brain cells and altering biochemical processes of the nervous system. This research presents a novel approach for future molecular studies using brain primary cells as model to screen pharmaceuticals and environmental agents for risk assessment.