Characterizing variability in total mercury hair:blood ratio in the general Canadian population.

BACKGROUND/OBJECTIVES: The body burden of mercury in humans can be measured through hair or blood biomarkers. To compare results from different studies, it is often required to convert mercury in hair to an equivalent level in blood, using a default hair:blood ratio of 250:1 by the World Health Organization (WHO). However, the actual ratio may vary within and between populations. The objectives of this study were to analyze the hair:blood mercury ratio in the general Canadian population, explore factors associated with higher/lower ratios, and determine if the standard ratio of 250:1 is supported. METHODS: The Canadian Health Measures Survey (CHMS) Cycle 5 (2016-2017) measured total mercury (THg) in both hair and blood of 1168 participants 20-59 years of age. We calculated geometric mean (GM) concentrations of THg for this entire sample and subgroups. The subgroups included biological sex, women of childbearing age, race, hair treatments, categories of blood and hair selenium, urinary arsenobetaine/arsenocholine, categories of blood and hair mercury, and food consumption. We calculated a hair:blood ratio for each participant and determined population-level ratios from the GMs of the distributions. Differences by subgroups, and agreement with the WHO ratio of 250:1, were tested. The combined effect of factors on the THg hair:blood ratio was explored using staged regression analysis. RESULTS: For participants with paired hair and blood mercury measurements, the GM of the hair:blood THg ratio was 293 (95%CI:273-316), and significantly >250. In women of childbearing age, the ratio did not differ from 250. The GMs of the ratio were higher (i.e.>300) for second tertile blood selenium (365, 95%CI:307-433), third and fourth quartiles hair mercury (347, 95%CI:308-390 and 376, 95%CI:336-422), and consumers of shellfish (338, 95%CI:308-371). Shellfish consumption was the only statistically significant factor associated with the hair:blood ratio as identified in the regression model. CONCLUSIONS: The mean hair:blood THg ratio among Canadians generally exceeded the default ratio of 250:1. Higher ratios were observed in certain subgroups, such as seafood consumers, and shellfish consumption was the most important variable associated with the ratio. Our results suggest that population-specific hair:blood THg ratios be considered, if possible, when converting mercury levels from hair to blood to better characterize the variation around the conversion.

Localizing organomercury uptake and accumulation in zebrafish larvae at the tissue and cellular level.

Using synchrotron x-ray fluorescence mapping, we have examined the uptake and localization of organic mercury in zebrafish larvae. Strikingly, the greatest accumulation of methyl and ethyl mercury compounds was highly localized in the rapidly dividing lens epithelium, with lower levels going to brain, optic nerve, and various other organs. The data suggest that the reported impairment of visual processes by mercury may arise not only from previously reported neurological effects, but also from direct effects on the ocular tissue. This novel approach is a powerful tool for directly investigating the molecular toxicology of heavy metals, and should be equally applicable to the study of a wide range of elements in developing embryos.

The heat-inducible zebrafish hsp70 gene is expressed during normal lens development under non-stress conditions.

In the present study, we show that the stress-inducible hsp70 gene in zebrafish is strongly and specifically expressed during normal lens formation from 28 to 38 hours post-fertilization, and is subsequently downregulated by 2 days of age. Only weak constitutive hsp70 mRNA signal was sporadically observed in other embryonic tissues. Similarly, transgenic fish carrying a 1.5 kb fragment of the hsp70 promoter linked to eGFP exhibited fluorescence only in the lens. In contrast, both the endogenous hsp70 gene and the transgene were strongly expressed throughout the embryo following heat shock at the same developmental stages.

Developmental toxicology of cadmium in living embryos of a stable transgenic zebrafish line.

The toxic effects of cadmium and other heavy metals have been well established, and many of these and other environmental pollutants are known to be embryotoxic or teratogenic. However, it has proven difficult to identify individual cells that respond to toxicants among the wide range of cell populations in an intact animal, particularly during early development when cells are continually changing their molecular and physiologic characteristics as they differentiate. Here we report the establishment of an in vivo system that uses hsp70 gene activation as a measure of cadmium toxicity in living early larvae of transgenic zebrafish carrying a stably integrated hsp70-enhanced green fluorescent protein (eGFP) reporter gene. We demonstrate that eGFP expression in this strain of fish acts as an accurate and reproducible indicator of cell-specific induction of hsp70 gene expression. Furthermore, the transgene responds in a dose-dependent manner at concentrations similar to those observed for morphologic indicators of early-life-stage toxicity and is sensitive enough to detect cadmium at doses below the median combined adverse effect concentration and the median lethal concentration. The stable nature of this transgenic line should allow for extremely rapid and reproducible toxicologic profiling of embryos and larvae throughout development.

Brief embryonic cadmium exposure induces a stress response and cell death in the developing olfactory system followed by long-term olfactory deficits in juvenile zebrafish.

The toxic effects of cadmium and other metals have been well established. A primary target of these metals is known to be the olfactory system, and fish exposed to a number of different waterborne metals display deficiencies in olfaction. Importantly, exposure over embryonic/larval development periods can cause deficits in chemosensory function in juvenile fish, but the specific cell types affected are unknown. We have previously characterized a transgenic zebrafish strain expressing the green fluorescent protein (eGFP) gene linked to the hsp70 gene promoter, and shown it to be a useful tool for examining cell-specific toxicity in living embryos and larvae. Here we show that the hsp70/eGFP transgene is strongly and specifically upregulated within the olfactory sensory neurons (OSNs) of transgenic zebrafish larvae following a brief 3-h exposure to water-borne cadmium. This molecular response was closely correlated to an endpoint for tissue damage within the olfactory placode, namely cell death. Furthermore, cadmium-induced olfactory cytotoxicity in zebrafish larvae gives rise to more permanent effects. Juvenile zebrafish briefly exposed to cadmium during early larval development display deficits in olfactory-dependent predator avoidance behaviors 4-6 weeks after a return to clean water. Lateral line neuromasts of exposed zebrafish larvae also activate both the endogenous hsp70 gene and the hsp70/eGFP transgene. The data reveal that even a very brief exposure period that gives rise to cell death within the developing olfactory placode results in long-term deficits in olfaction, and that hsp70/eGFP may serve as an effective indicator of sublethal cadmium exposure in sensory cells.

Heat shock gene expression and function during zebrafish embryogenesis.

Recent work in the zebrafish, Danio rerio, indicates that heat shock genes are expressed in unique spatial patterns under non-stress conditions. In particular, hsp90alpha is expressed during the normal differentiation of striated muscle fibres, and hsp70-4 is expressed during normal lens development in the eye. Furthermore, disruption of the activity of either of these genes or their protein products gives rise to unique embryonic phenotypes that result from failures in proper somitic muscle development and lens development, respectively. Embryonic hsp70-4 expression is also activated in a cell-specific manner following heavy metal exposure. This has allowed for the development of a hsp70-4/eGFP reporter gene system in stable transgenic zebrafish that serves as a reliable yet extremely quick indicator of cell-specific toxicity in the context of the multicellular, living embryo.