Regulation of MicroRNAs After Spinal Cord Injury in Adult Zebrafish.

Spinal cord injury (SCI) is a central nerve injury that often leads to loss of motor and sensory functions at or below the level of the injury. Zebrafish have a strong ability to repair after SCI, but the role of microRNAs (miRNAs) after SCI remains unclear. Locomotor behavior analysis showed that adult zebrafish recovered about 30% of their motor ability at 2 weeks and 55% at 3 weeks after SCI, reflecting their strong ability to repair SCI. Through miRNA sequencing, mRNA sequencing, RT-qPCR experiment verification, and bioinformatics predictive analysis, the key miRNAs and related genes in the repair of SCI were screened. A total of 38 miRNAs were significantly different, the top ten miRNAs were verified by RT-qPCR. The prediction target genes were verified by the mRNAs sequencing results at the same time point. Finally, 182 target genes were identified as likely to be networked regulated by the 38 different miRNAs. GO and KEGG enrichment analysis found that miRNAs targeted gene regulation of many key pathways, such as membrane tissue transport, ribosome function, lipid binding, and peroxidase activity. The PPI network analysis showed that miRNAs were involved in SCI repair through complex network regulation, among which dre-miR-21 may enhance cell reversibility through nop56, and that dre-miR-125c regulates axon growth through kpnb1 to repair SCI.

Locomotor Assays in Drosophila Larvae and Adult Flies.

Brain defects often lead to motor dysfunctions in humans. Drosophila melanogaster has been one of the most useful organisms in the study of neuronal biology due to its similarities with humans and has contributed to a more detailed understanding of the effects of genetic dysfunctions in the brain on behavior. We herein present modified protocols for the crawling assay with larvae and the climbing assay with adult flies that are simple to perform as well as a series of commands for ImageJ to automatically analyze data for the crawling assay.

In vivo evaluation of the neurogenotoxic effects of exposure to validamycin A in neuroblasts of Drosophila melanogaster larval brain.

Agriculture commonly utilizes crop protection products to tackle infestations from fungi, parasites, insects, and weeds. Validamycin A, an inhibitor of trehalase, possesses antibiotic and antifungal attributes. Epidemiological evidence has led to concerns regarding a potential link between pesticide usage and neurodegenerative diseases. The fruit fly, Drosophila melanogaster, has been recognized as a reliable model for genetic research due to its significant genetic similarities with mammals. Here, we propose to use D. melanogaster as an effective in vivo model system to investigate the genotoxic risks associated with exposure to validamycin A. In this study, we performed a neurotoxic evaluation of validamycin A in D. melanogaster larvae. Several endpoints were evaluated, including toxicity, intracellular oxidative stress (reactive oxygen species), intestinal damage, larval behavior (crawling behavior, light/dark sensitivity assay, and temperature sensitivity assay), locomotor (climbing) behavior, and neurogenotoxic effects (impaired DNA via Comet assay, enhanced by Endo III and formamidopyrimidine DNA glycosylase [FPG]). The results showed that exposure to validamycin A, especially at higher doses (1 and 2.5 mM), induced DNA impairment in neuroblasts as observed by Comet assay. Both larvae and adults exhibited behavioral changes and produced reactive oxygen species. Most importantly, this research represents a pioneering effort to report neurogenotoxicity data specifically in Drosophila larval neuroblasts, thus underscoring the importance of this species as a testing model in exploring the biological impacts of validamycin A. The in vivo findings from the experiments are a valuable and novel addition to the existing validamycin A neurogenotoxicity database.

Minor metabolomic disturbances induced by glyphosate-isopropylammonium exposure at environmentally relevant concentrations in an aquatic turtle, Pelodiscus sinensis.

The ecotoxicological and environmental impacts of glyphosate-based herbicides have received considerable attention due to their extensive use globally. However, the potential for adverse effects in cultured non-fish vertebrate species are commonly ignored. In this study, effects on growth, indicators of functional performance, gut microbial diversity, liver antioxidant responses and metabolite profiles were evaluated in soft-shelled turtle hatchlings (Pelodiscus sinensis) exposed to different concentrations of glyphosate-isopropylammonium (0, 0.02, 0.2, 2 and 20 mg/L). No significant changes in growth or functional performance (food intake, swimming speed), gut microbiota, and liver antioxidant responses (SOD and CAT activities, MDA content) were observed in exposed turtles. However, hepatic metabolite profiles revealed distinct perturbations that primarily involved amino acid metabolism in turtles exposed to environmentally relevant concentrations. Overall, our results suggested that metabolite profiles may be more sensitive than phenotypic or general physiological endpoints and gut microbiota profiling, and indicate a potential mechanism of hepatotoxicity caused by glyphosate-isopropylammonium based on untargeted metabolomics analysis. Furthermore, the toxicity of glyphosate at environmentally relevant concentrations might be relatively minor in aquatic turtle species.

Hazard Assessment of the Effects of Acute and Chronic Exposure to Permethrin, Copper Hydroxide, Acephate, and Validamycin Nanopesticides on the Physiology of Drosophila: Novel Insights into the Cellular Internalization and Biological Effects.

New insights into the interactions between nanopesticides and edible plants are required in order to elucidate their impacts on human health and agriculture. Nanopesticides include formulations consisting of organic/inorganic nanoparticles. Drosophila melanogaster has become a powerful model in genetic research thanks to its genetic similarity to mammals. This project mainly aimed to generate new evidence for the toxic/genotoxic properties of different nanopesticides (a nanoemulsion (permethrin nanopesticides, 20 ± 5 nm), an inorganic nanoparticle as an active ingredient (copper(II) hydroxide [Cu(OH)2] nanopesticides, 15 ± 6 nm), a polymer-based nanopesticide (acephate nanopesticides, 55 ± 25 nm), and an inorganic nanoparticle associated with an organic active ingredient (validamycin nanopesticides, 1177 ± 220 nm)) and their microparticulate forms (i.e., permethrin, copper(II) sulfate pentahydrate (CuSO4·5H2O), acephate, and validamycin) widely used against agricultural pests, while also showing the merits of using Drosophila-a non-target in vivo eukaryotic model organism-in nanogenotoxicology studies. Significant biological effects were noted at the highest doses of permethrin (0.06 and 0.1 mM), permethrin nanopesticides (1 and 2.5 mM), CuSO4·5H2O (1 and 5 mM), acephate and acephate nanopesticides (1 and 5 mM, respectively), and validamycin and validamycin nanopesticides (1 and 2.5 mM, respectively). The results demonstrating the toxic/genotoxic potential of these nanopesticides through their impact on cellular internalization and gene expression represent significant contributions to future nanogenotoxicology studies.

How do the physiological traits of a lizard change during its invasion of an oceanic island?

Physiology is crucial for the survival of invasive species in new environments. Yet, new climatic conditions and the limited genetic variation found within many invasive populations may influence physiological responses to new environmental conditions. Here, we studied the case of the delicate skinks (Lampropholis delicata) invading Lord Howe Island (LHI), Australia. On LHI, the climate is different from the mainland source of the skinks, and independent introduction events generated invasive populations with distinct genetic backgrounds. To understand how climate and genetic background may shape physiological responses along biological invasions, we compared the physiological traits of a source and two invasive (single-haplotype and multi-haplotype) populations of the delicate skink. For each population, we quantified physiological traits related to metabolism, sprint speed, and thermal physiology. We found that, for most physiological traits analysed, population history did not influence the ecophysiology of delicate skinks. However, invasive populations showed higher maximum speed than the source population, which indicates that locomotor performance might be a trait under selection during biological invasions. As well, the invasive population with a single haplotype was less cold-tolerant than the multi-haplotype and source populations. Our results suggest that limited genetic variability and climate may influence physiological responses of invasive organisms in novel environments. Incorporating the interplay between genetic and physiological responses into models predicting species invasions can result in more accurate understanding of the potential habitats those species can occupy.

Effects of extremely low-frequency electromagnetic field on different developmental stages of Drosophila melanogaster.

PURPOSE: The model biological organism Drosophila melanogaster has been utilized to assess the effect of extremely low-frequency electromagnetic field (ELF-EMF) on locomotion, longevity, developmental dynamics, cell viability and oxidative stress. MATERIALS AND METHOD: Developmental stages of Drosophila melanogaster (Oregon R strain) individually exposed to ELF-EMF (75 Hz, 550 µT) for 6 h once for acute exposure. For chronic exposure, complete life cycle of fly, that is, egg to adult fly was exposed to ELF-EMF for 6 h daily. The effect of exposure on their crawling and climbing ability, longevity, development dynamics, cellular damage and oxidative stress (generation of reactive oxygen species (ROS)) was evaluated. RESULTS: The crawling ability of larvae was significantly (p < .05) reduced on acute (third stage instar larvae) as well as chronic exposure (F0 and F1 larvae). When locomotion of flies was tested using climbing assay, no alteration was observed in their climbing ability under both acute and chronic exposure; however, when their speed of climbing was compared, a significant decrease in speed of F1 flies was observed (p = .0027) on chronic exposure. The survivability of flies was significantly affected under chronic and acute exposure (at third stage instar larvae). In case of acute exposure of the third stage instar larvae, although all the flies were eclosed by the 17th day, there was a significant decline in the number of flies (p = .007) in comparison to control. While in case of chronic exposure apart from low number of flies eclosed in comparison to control, there was delay in eclosion by one day (p = .0004). Using trypan blue assay, the internal gut damage of third stage instar larvae was observed. Under acute exposure condition at third stage instar larvae, 30% larvae has taken up trypan blue, while only 10% larvae from acute exposure at adult stage. On chronic exposure, 50% larvae of the F1 generation have taken up trypan blue. On evaluation of oxidative stress, there is a significant rise in ROS in case of acute exposure at third stage instar larvae (p = .0004), adult fly stage (p = .0004) and chronic exposure (p = .0001). CONCLUSION: ELF-EMF has maximum effects on acute exposure of third stage instar larvae and chronic exposure (egg to adult fly stage). These results suggest that electromagnetic radiations, though, have become indispensible part of our lives but they plausibly affect our health.

Shexiang Baoxin Pill, a Proprietary Multi-Constituent Chinese Medicine, Prevents Locomotor and Cognitive Impairment Caused by Brain Ischemia and Reperfusion Injury in Rats: A Potential Therapy for Neuropsychiatric Sequelae of Stroke.

Ischemic stroke is a common type of cerebrovascular event and also the leading cause of disability. Post-stroke cognitive impairment occurs frequently in stroke survivors. Shexiang Baoxin Pill (SBP) is a proprietary Chinese medicine, initially used to treat cardiovascular diseases. Herein, we aim to explore the effects of SBP on oxygen glucose deprivation and reoxygenation (OGD/R) in neuronal cells (CATH.a) and cerebral ischemia/reperfusion injury induced post-stroke cognitive impairment in middle cerebral artery occlusion (MCAO) rat model. MCAO rats received two doses of oral SBP treatment (28 or 56 mg/kg) after 1 h of operation and once daily for 2 weeks continuously. Behavioral tests, immunoblotting, and immunofluorescence were examined after 14 days. Current data suggest that SBP enhanced cell viability and downregulated apoptosis via activating the PI3K/Akt signaling pathway in CATH. a cells. Furthermore, 14 days of SBP treatment promoted the recovery of learning and locomotor function in the MCAO rats. SBP up-regulated the expression of p-Akt, p-GSK3ß, as well as the expression of NMDAR1, PSD-95, and AMPAR. Also, SBP down-regulated the expression of p-CaMKII. These results indicated that long-term SBP treatment might be a potential option for cognitive impairment induced by the ischemic stroke.

Thermal physiological performance of two freshwater turtles acclimated to different temperatures.

The thermal physiological performance of invasive species may play a crucial role in determining their invasion success. In this study, we acclimated two cohorts of hatchlings of freshwater turtles (native Mauremys reevesii and invasive Trachemys scripta elegans) from low and high-latitude collection sites, respectively, to different thermal conditions (20 and 30 °C) for 4 weeks, and then compared their thermal tolerance and locomotor performance. T. scripta elegans hatchlings could swim faster (but righted themselves more slowly), and tolerate a higher temperature and wider temperature range than M. reevesii hatchlings. Similarly, T. scripta elegans hatchlings had a greater maximal performance (Pmax) value for swimming speed (but a lower Pmax value for righting time) than M. reevesii hatchlings. Temperature acclimation had a significant impact on the thermal tolerance and locomotor ability of turtles, but the acclimation effect did not differ between the two species. T. scripta elegans hatchlings seemed to have a greater thermal plasticity than M. reevesii hatchlings. High-latitude individuals showed a greater low-temperature tolerance, but lower locomotor ability (longer righting time) than low-latitude ones. However, the thermal plasticity did not differ between latitudinal cohorts. Our results indicated that T. scripta elegans performed better than M. reevesii, which might contribute to its range expansion and invasive success.

Reduction in locomotor ability as a cost of reproduction in gravid snakes.

Recent studies suggest that lower survival among gravid squamate reptiles may be partially the result of decreased locomotor ability during gestation. In this study, we compared the speed and endurance of female garter snakes (Thamnophis marcianus), before, during, and after pregnancy. Gravid snakes had significantly lower locomotor performance than did non-gravid females, and performance varied among stages of gestation, reaching a minimum 0-6 weeks prior to parturition. Both number of offspring and relative clutch mass were inversely correlated with locomotor performance; as females increased these traits, locomotor ability decreased. If reduced locomotor performance results in greater risk of predation and/or lowered foraging ability, then natural selection (operating via differential mortality or feeding rates of gravid females) may result in important constraints on both clutch size and relative clutch mass in squamates.