Elizabeth McCandlish

Neural stem cell apoptosis after low‐methylmercury exposures in postnatal hippocampus produce persistent cell loss and adolescent memory deficits

The developing brain is particularly sensitive to exposures to environmental contaminants. In contrast to the adult, the developing brain contains large numbers of dividing neuronal precursors, suggesting that they may be vulnerable targets. The postnatal day 7 (P7) rat hippocampus has populations of both mature neurons in the CA1–3 region as well as neural stem cells (NSC) in the dentate gyrus (DG) hilus, which actively produce new neurons that migrate to the granule cell layer (GCL). Using this well‐characterized NSC population, we examined the impact of low levels of methylmercury (MeHg) on proliferation, neurogenesis, and subsequent adolescent learning and memory behavior. Assessing a range of exposures, we found that a single subcutaneous injection of 0.6 µg/g MeHg in P7 rats induced caspase activation in proliferating NSC of the hilus and GCL. This acute NSC death had lasting impact on the DG at P21, reducing cell numbers in the hilus by 22% and the GCL by 27%, as well as reductions in neural precursor proliferation by 25%. In contrast, non‐proliferative CA1–3 pyramidal neuron cell number was unchanged. Furthermore, animals exposed to P7 MeHg exhibited an adolescent spatial memory deficit as assessed by Morris water maze. These results suggest that environmentally relevant levels of MeHg exposure may decrease NSC populations and, despite ongoing neurogenesis, the brain may not restore the hippocampal cell deficits, which may contribute to hippocampal‐dependent memory deficits during adolescence.

Functional Rehabilitation of Cadmium-Induced Neurotoxicity Despite Persistent Peripheral Pathophysiology in the Olfactory System

Intranasal exposure to the heavy metal cadmium has been linked to olfactory dysfunction and neurotoxicity. Here, we combine optical imaging of in vivo neurophysiology, genetically defined anatomical tract tracing, mass spectrometry, and behavioral psychophysical methods to evaluate the persistent harmful effects of acute intranasal exposure to cadmium in a mouse model and to investigate the functional consequences of sensory rehabilitation training. We find that an acute intranasal instillation of cadmium chloride leads to an accumulation of cadmium in the brains olfactory bulb that persists for at least 4 weeks. This is accompanied by persistent severe pathophysiology of the olfactory nerve, a gradual reduction in axonal projections from the olfactory epithelium, and complete impairment on an olfactory detection task. Remarkably, 2 weeks of odorant-guided operant conditioning training proved sufficient to restore olfactory detection performance to control levels in cadmium-exposed mice. Optical imaging from rehabilitated mice showed that this training did not cause any detectable restoration of olfactory nerve function, suggesting that the recovery of function was mediated by central neuroplasticity in which the brain learned to interpret the degraded sensory input. These data demonstrate that sensory learning can mask even severe damage from neurotoxicants and suggest that explicit sensory training may be useful in rehabilitation of olfactory dysfunction.

N-acetyl cysteine treatment reduces mercury-induced neurotoxicity in the developing rat hippocampus

Mercury is an environmental toxicant that can disrupt brain development. However, although progress has been made in defining its neurotoxic effects, we know far less about available therapies that can effectively protect the brain in exposed individuals. We previously developed an animal model in which we defined the sequence of events underlying neurotoxicity: Methylmercury (MeHg) injection in postnatal rat acutely induced inhibition of mitosis and stimulated apoptosis in the hippocampus, which later resulted in intermediate‐term deficits in structure size and cell number. N‐acetyl cysteine (NAC) is the N‐acetyl derivative of L‐cysteine used clinically for treatment of drug intoxication. Here, based on its known efficacy in promoting MeHg urinary excretion, we evaluated NAC for protective effects in the developing brain. In immature neurons and precursors, MeHg (3 μM) induced a >50% decrease in DNA synthesis at 24 hr, an effect that was completely blocked by NAC coincubation. In vivo, injection of MeHg (5 μg/g bw) into 7‐day‐old rats induced a 22% decrease in DNA synthesis in whole hippocampus and a fourfold increase in activated caspase‐3‐immunoreactive cells at 24 hr and reduced total cell numbers by 13% at 3 weeks. Treatment of MeHg‐exposed rats with repeated injections of NAC abolished MeHg toxicity. NAC prevented the reduction in DNA synthesis and the marked increase in caspase‐3 immunoreactivity. Moreover, the intermediate‐term decrease in hippocampal cell number provoked by MeHg was fully blocked by NAC. Altogether these results suggest that MeHg toxicity in the perinatal brain can be ameliorated by using NAC, opening potential avenues for therapeutic intervention.

Transgenic Expression of the Human MRP2 Transporter Reduces Cisplatin Accumulation and Nephrotoxicity in Mrp2-Null Mice

The chemotherapeutic drug cisplatin is actively transported into proximal tubules, leading to acute renal injury. Previous studies suggest that the multidrug resistance-associated protein 2 (Mrp2) transporter may efflux cisplatin conjugates from cells. We sought to determine whether the absence of Mrp2 alters the accumulation and toxicity of platinum in the kidneys of mice and whether transgenic expression of the human MRP2 gene could protect against cisplatin injury in vivo. Plasma, kidneys, and livers from vehicle- and cisplatin-treated wild-type and Mrp2-null mice were collected for quantification of platinum and toxicity. By 24 hours, twofold higher concentrations of platinum were detected in the kidneys and livers of Mrp2-null mice compared with wild types. Enhanced platinum concentrations in Mrp2-null mice were observed in DNA and cytosolic fractions of the kidneys. Four days after cisplatin treatment, more extensive proximal tubule injury was observed in Mrp2-null mice compared with wild-type mice. Kidneys from naive Mrp2-null mice had elevated glutathione S-transferase mRNA levels, which could increase the formation of cisplatin-glutathione conjugates that may be metabolized to toxic thiol intermediates. Transgenic expression of the human MRP2 gene in Mrp2-null mice reduced the accumulation and nephrotoxicity of cisplatin to levels observed in wild-type mice. These data suggest that deficiency in Mrp2 lowers platinum excretion and increases susceptibility to kidney injury, which can be rescued by the human MRP2 ortholog.

Hippocampal developmental vulnerability to methylmercury extends into prepubescence.

The developing brain is sensitive to environmental toxicants such as methylmercury (MeHg), to which humans are exposed via contaminated seafood. Prenatal exposure in children is associated with learning, memory and IQ deficits, which can result from hippocampal dysfunction. To explore underlying mechanisms, we have used the postnatal day (P7) rat to model the third trimester of human gestation. We previously showed that a single low exposure (0.6 μg/gbw) that approaches human exposure reduced hippocampal neurogenesis in the dentate gyrus (DG) 24 h later, producing later proliferation and memory deficits in adolescence. Yet, the vulnerable stem cell population and period of developmental vulnerability remain undefined. In this study, we find that P7 exposure of stem cells has long-term consequences for adolescent neurogenesis. It reduced the number of mitotic S-phase cells (BrdU), especially those in the highly proliferative Tbr2+ population, and immature neurons (Doublecortin) in adolescence, suggesting partial depletion of the later stem cell pool. To define developmental vulnerability to MeHg in prepubescent (P14) and adolescent (P21) rats, we examined acute 24 h effects of MeHg exposure on mitosis and apoptosis. We found that low exposure did not adversely impact neurogenesis at either age, but that a higher exposure (5 μg/gbw) at P14 reduced the total number of neural stem cells (Sox2+) by 23% and BrdU+ cells by 26% in the DG hilus, suggesting that vulnerability diminishes with age. To determine whether these effects reflect changes in MeHg transfer across the blood brain barrier (BBB), we assessed Hg content in the hippocampus after peripheral injection and found that similar levels (~800 ng/gm) were obtained at 24 h at both P14 and P21, declining in parallel, suggesting that changes in vulnerability depend more on local tissue and cellular mechanisms. Together, we show that MeHg vulnerability declines with age, and that early exposure impairs later neurogenesis in older juveniles.

Risk Assessment for Children Exposed to Beach Sands Impacted by Oil Spill Chemicals

Due to changes in the drilling industry, oil spills are impacting large expanses of coastlines, thereby increasing the potential for people to come in contact with oil spill chemicals. The objective of this manuscript was to evaluate the health risk to children who potentially contact beach sands impacted by oil spill chemicals from the Deepwater Horizon disaster. To identify chemicals of concern, the U.S. Environmental Protection Agency’s (EPA’s) monitoring data collected during and immediately after the spill were evaluated. This dataset was supplemented with measurements from beach sands and tar balls collected five years after the spill. Of interest is that metals in the sediments were observed at similar levels between the two sampling periods; some differences were observed for metals levels in tar balls. Although PAHs were not observed five years later, there is evidence of weathered-oil oxidative by-products. Comparing chemical concentration data to baseline soil risk levels, three metals (As, Ba, and V) and four PAHs (benzo[a]pyrene, benz[a]anthracene, benzo[b]fluoranthene, and dibenz[a,h]anthracene) were found to exceed guideline levels prompting a risk assessment. For acute or sub-chronic exposures, hazard quotients, computed by estimating average expected contact behavior, showed no adverse potential health effects. For cancer, computations using 95% upper confidence limits for contaminant concentrations showed extremely low increased risk in the 10−6 range for oral and dermal exposure from arsenic in sediments and from dermal exposure from benzo[a]pyrene and benz[a]anthracene in weathered oil. Overall, results suggest that health risks are extremely low, given the limitations of available data. Limitations of this study are associated with the lack of toxicological data for dispersants and oil-spill degradation products. We also recommend studies to collect quantitative information about children’s beach play habits, which are necessary to more accurately assess exposure scenarios and health risks.

Concentrations of heavy metals in fish species targeted by anglers in central New Jersey: A pilot study

ABSTRACT The purpose of this study was to investigate the concentrations of toxic metals in the edible portion of fish tissue obtained from the Raritan River in New Brunswick, New Jersey (NJ) between April and May of 2014. Species collected for this study included commonly caught fish such as bluegill, small and large mouth bass, brown and rainbow trout, bullhead catfish, and white perch. Samples were filleted and the muscle tissue subsequently dehydrated and then chemically digested. Samples were analyzed using Gas chromatography mass spectrometry (GC–MS). Levels of 28 different metals were quantified for each specimen. In general, metal contamination in fish tissues was below the recommended limits. However, lead was found in one sample at a tissue concentration of 88 µg per 225 g fillet, which is above the recommended daily consumption limit as set by the Food and Safety Authority of Ireland. The maximum level found for arsenic was 23 µg per 225 g fillet. The fish samples taken from the Raritan River in New Brunswick, NJ for this study did not contain dangerous levels of most of the metals tested.