Characterization of the avian aryl hydrocarbon receptor 1 from blood using non-lethal sampling methods
The amino acid sequence of the aryl hydrocarbon receptor 1 ligand binding domain (AHR1 LBD) is an important determinant of sensitivity to dioxin-like compounds in avian species. We are interested in surveying AHR1 LBD sequences in a large number of birds as a means of identifying species that are particularly sensitive to dioxin-like compounds. Our original method for determining AHR1 LBD genotype used liver tissue and required lethal sampling. Here we present two alternate methods for determining AHR1 LBD genotype which use non-lethal sampling and are more appropriate for ecologically sensitive species. First, we establish that AHR1 LBD mRNA is expressed in avian blood and test a variety of blood collection and handling protocols in order to establish a method that is convenient for field collections. Our findings also identify which types of archival blood samples might be appropriate for AHR1 LBD sequence determination. Second, we present a method for obtaining AHR1 LBD coding sequences from DNA. A DNA-based method is advantageous because DNA can be isolated from many tissue types, is more stable than RNA, and requires less specific sample handling and preservation. This work extends applicability of a genetic screen for dioxin sensitivity to a larger number of species and sample types including endangered species and potentially museum specimens.
KeywordsAvian Dioxin-like compounds Aryl hydrocarbon receptor 1 Species sensitivity Non-lethal sampling
We thank Ian Ritchie for facilitating collection of the American kestrel samples. We are grateful to Guy Morrison for donation of ivory gull samples and to Ryan Zimmerling for donation of piping plover samples. This work was supported by Environment Canada’s Wildlife Toxicology and Disease Program and by Environment Canada’s Strategic Technology Applications of Genomics for the Environment (STAGE) program.
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