Development of gene expression-based biomarkers of exposure to metals and pesticides in the freshwater amphipod Hyalella azteca

Ecological risk assessment (ERA) is a framework for monitoring risks of exposure and adverse effects of environmental stressors to populations or communities of interest. One tool of ERA is the biomarker, which is a characteristic of an organism that reliably indicates exposure to or effects of a stressor like chemical pollution. Traditional biomarkers which rely on characteristics at the tissue level and higher often detect only acute exposures to stressors. Sensitive molecular biomarkers may detect lower stressor levels than traditional biomarkers, which helps inform risk mitigation and restoration efforts before populations and communities are irreversibly affected. In this study I developed gene expression-based molecular biomarkers of exposure to metals and insecticides in the model toxicological freshwater amphipod Hyalella azteca. My goals were to not only create sensitive molecular biomarkers for these chemicals, but also to show the utility and versatility of H. azteca in molecular studies for toxicology and risk assessment. I sequenced and assembled the H. azteca transcriptome to identify reference and stress-response gene transcripts suitable for expression monitoring. I exposed H. azteca to sub-lethal concentrations of metals (cadmium and copper) and insecticides (DDT, permethrin, and imidacloprid). Reference genes used to create normalization factors were determined for each exposure using the programs BestKeeper, GeNorm, and NormFinder. Both metals increased expression of a nuclear transcription factor (Cnc), an ABC transporter (Mrp4), and a heat shock protein (Hsp90), giving evidence of general metal exposure signature. Cadmium uniquely increased expression of a DNA repair protein (Rad51) and increased Mrp4 expression more than copper (7-fold increase compared to 2-fold increase). Together these may be unique biomarkers distinguishing cadmium and copper exposures. DDT increased expression of Hsp90, Mrp4, and the immune response gene Lgbp. Permethrin increased expression of a cytochrome P450 (Cyp2j2) and decreased expression of the immune response gene Lectin-1. Imidacloprid did not affect gene expression. Unique biomarkers were seen for DDT and permethrin, but the genes studied were not sensitive enough to detect imidacloprid at the levels used here. I demonstrated that gene expression in H. azteca detects specific chemical exposures at sub-lethal concentrations, making expression monitoring using this amphipod a useful and sensitive biomarker for risk assessment of chemical exposure.

Cortisol Reactivity and Observed Parenting among Mothers of Children with and without ADHD

Parenting is a robust predictor of developmental outcomes among children with ADHD. Early parenting predicts the persistence and course of ADHD and comorbid problems above and beyond risk associated with shared genetic effects. Yet, on average, mothers of children with ADHD are less positive and more negative in their parent-child interactions compared to mothers of non-disordered children. Little is known about psychobiological markers which may be associated with individual variations in maternal parenting in families of children with ADHD. Neurobiological models of parenting suggest that maternal cortisol levels following a stressor may be positively associated with hostile and intrusive parenting; however, to date no studies have examined maternal cortisol reactivity and parenting in school-age, or clinical samples of, children. Mothers’ regulation of physiological stress responses may be particularly important for families of children with ADHD, as parenting a child with chronically challenging behaviors represents a persistent environmental stressor. The current study sought to extend the existing literature by providing an empirical examination of the relationship between maternal cortisol reactivity following two laboratory stressors and parenting among mothers of children with and without ADHD. It was hypothesized that child ADHD group would moderate the relationship between cortisol reactivity and self-reported and observed parenting. Greater total cortisol output and greater increase in cortisol during the TSST were associated with decreased positive parenting and increased negative and directive parenting, with the exception of parental involvement, which was associated with increased cortisol output during the TSST. Conversely, cortisol output during the PCI was associated with increased positive parenting, increased parental involvement, and decreased negative parenting. In contrast to the TSST, a greater decrease in cortisol during the PCI indicated more positive parenting and parental involvement. These associations were specific to mothers of children with ADHD, with the exception of maternal directiveness, which was specific to comparison mothers. Findings add to our understanding of physiological processes associated with maternal parenting and contribute to an integrative biological, psychological, and cognitive process model of parenting in families of children with ADHD.