Temporal sex specific brain gene expression pattern during early rat embryonic development.

Background: The classical concept of brain sex differentiation suggests that steroid hormones released from the gonads program male and female brains differently. However, several studies indicate that steroid hormones are not the only determinant of brain sex differentiation and that genetic differences could also be involved. Methods: In this study, we have performed RNA sequencing of rat brains at embryonic days 12 (E12), E13, and E14. The aim was to identify differentially expressed genes between male and female rat brains during early development. Results: Analysis of genes expressed with the highest sex differences showed that Xist was highly expressed in females having XX genotype with an increasing expression over time. Analysis of genes expressed with the highest male expression identified three early genes, Sry2, Eif2s3y, and Ddx3y. Discussion: The observed sex-specific expression of genes at early development confirms that the rat brain is sexually dimorphic prior to gonadal action on the brain and identifies Sry2 and Eif2s3y as early genes contributing to male brain development.

Assessing organism differences in mixed metal sensitivity.

Metal contamination of aquatic environments remains a major concern and has received significant attention in recent years. The present study aimed to evaluate the effects of metal mixtures of varying concentrations over time in a lake receiving runoff water from a decommissioned mine. By subjecting several organisms to this water, we aimed to identify the most susceptible species, thus enabling a comprehensive evaluation of the risk posed by different toxins to the biotic environment. We have evaluated the effects of mixed metal exposure on survival and stress gene expression in selected invertebrate and vertebrate model species. Our observations revealed differences in sensitivity among the invertebrate models Caenorhabditis elegans, Daphnia magna, Ceriodaphnia dubia, and Heterocypris incongruens, as well as in the vertebrate model Zebrafish (Danio rerio) and two cell lines; a zebrafish liver cell line (ZFL) and a human hepatocellular carcinoma cell line (HepG2). While the sensitivity shows great variation among the tested species, the expression of metallothionein was consistent with the levels of metals found in the mixed exposure media. Despite differences in acute toxicity, the universal induction of mt1/A and mt2/B genes make them important biomarkers for assessing the environmental risk of metals.

Utilization of Computer Classification Methods for Exposure Prediction and Gene Selection in Daphnia magna Toxicogenomics.

Zinc (Zn) is an essential element that influences many cellular functions. Depending on bioavailability, Zn can cause both deficiency and toxicity. Zn bioavailability is influenced by water hardness. Therefore, water quality analysis for health-risk assessment should consider both Zn concentration and water hardness. However, exposure media selection for traditional toxicology tests are set to defined hardness levels and do not represent the diverse water chemistry compositions observed in nature. Moreover, these tests commonly use whole organism endpoints, such as survival and reproduction, which require high numbers of test animals and are labor intensive. Gene expression stands out as a promising alternative to provide insight into molecular events that can be used for risk assessment. In this work, we apply machine learning techniques to classify the Zn concentrations and water hardness from Daphnia magna gene expression by using quantitative PCR. A method for gene ranking was explored using techniques from game theory, namely, Shapley values. The results show that standard machine learning classifiers can classify both Zn concentration and water hardness simultaneously, and that Shapley values are a versatile and useful alternative for gene ranking that can provide insight about the importance of individual genes.

Influence of water hardness on zinc toxicity in Daphnia magna.

Zinc is an essential trace metal required for the maintenance of multiple physiological functions. Due to this, organisms can experience both zinc deficiency and toxicity. Hardness is recognized as one of the main modifying physiochemical factors regulating zinc bioavailability. Therefore, the present study analyzed the effect of hardness on zinc toxicity using Daphnia magna. Endpoint parameters were acute-toxicity, development, reproduction, and expression data for genes involved in metal regulation and oxidative stress. In addition, the temporal expression profiles of genes during the initiation of reproduction and molting were investigated. Water hardness influenced the survival in response to exposures to zinc. A zinc concentration of 50 µg/l in soft water (50 mg CaCO3 /L) caused 73% mortality after 96 h exposure, whereas the same zinc concentration in the hardest water did not cause any significant mortality. Moreover, increasing water hardness from 100 to 200 mg CaCO3 /L resulted in a reduced number of offspring. Fecundity was higher at first brood for groups exposed to higher Zn concentrations. The survival data were used to assess the precision of the bioavailability models (Bio-met) and the geochemical model (Visual MINTEQ). As the Bio-met risk predictions overestimated the Zn toxicity, a competition-based model to describe the effects of hardness on zinc toxicity is proposed. This approach can be used to minimize differences in setting environmental quality standards. Moreover, gene expression data showed that using the toxicogenomic approach was more sensitive than the physiological endpoints. Therefore, data presented in the study can be used to improve risk assessment for zinc toxicity.

Sublethal effects of DBE-DBCH diastereomers on physiology, behavior, and gene expression of Daphnia magna.

1,2-dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH) is a brominated flame retardant used in commercial and industrial applications. The use of DBE-DBCH containing products has resulted in an increased release into the environment. However, limited information is available on the long-term effects of DBE-DBCH and its effects in aquatic invertebrates. Thus, the present study was aimed at determining how DBE-DBCH diastereomers (αß and γδ) affects aquatic invertebrates using Daphnia magna as a model organism. Survival, reproduction, feeding, swimming behavior and toxicogenomic responses to environmental relevant concentrations of DBE-DBCH were analyzed. Chronic exposure to DBE-DBCH resulted in decreased lifespan, and reduced fecundity. Expression of genes involved in reproductive processes, vtg1 and jhe, were also inhibited. DBE-DBCH also induced hypoxia by inhibiting the transcription of genes involved in heme biosynthesis and oxygen transport. Furthermore, DBE-DBCH also inhibited feeding resulting in emptiness of the alimentary canal. Increased expression of the stress response biomarkers was observed following DBE-DBCH exposure. In addition, DBE-DBCH diastereomers also altered the swimming behavior of Daphnia magna. The present study demonstrates that DBE-DBCH cause multiple deleterious effects on Daphnia magna, including effects on reproduction and hormonal systems. These endocrine disrupting effects are in agreement with effects observed on vertebrates. Furthermore, as is the case in vertebrates, DBE-DBCH γδ exerted stronger effects than DBE-DBCH αß on Daphnia magna. This indicate that DBE-DBCH γδ has properties making it more toxic to all so far studied animals than DBE-DBCH αß.