In vivo aflatoxin B1 metabolism and hepatic DNA adduction in zebrafish (Danio rerio).
The zebrafish (Danio rerio) is assuming prominence in developmental genetics research. By comparison, little is known of tumorigenesis and nothing is known of carcinogen metabolism in this species. This study evaluated the ability of zebrafish to metabolize a well-characterized human carcinogen, aflatoxin B1 (AFB1), to phase I and phase II metabolites and assessed hepatic AFB1-DNA adduction in vivo. Fish i.p. injected with 50-400 micrograms [3H]AFB1/kg body wt displayed a linear dose response for hepatic DNA binding at 24 hr. AFB1-DNA adduct levels among treatments showed no statistical difference over the period from 1 to 21 days after injection, suggesting poor adduct repair in this species. DNA binding in female fish was 1-7-fold higher than that in males (p < 0.01). An in vitro AFB1 metabolism assay verified that zebrafish liver extracts oxidize AFB1 to the 8,9-epoxide proximate electrophile (Km = 79.0 +/- 16.4 microM, Vmax = 11.7 +/- 1.4 pmol/min/mg protein at 28 degrees C). The excretion of AFB1 and its metabolites was also examined by HPLC. As is typical of other fish studied, major metabolites excreted were aflatoxicol (AFL) and aflatoxicol-glucuronide (AFL-g), followed by unreacted AFB1. AFL appeared as early as 5 min after injection, whereas AFL-g was a significant metabolite after 18 hr. This study shows that in vivo administration of AFB1 to zebrafish results in moderate adduction of the carcinogen to liver DNA and that zebrafish have the capacity for both phase I and phase II metabolism of AFB1. The approximate fourfold difference between rainbow trout and zebrafish AFB1-DNA covalent binding index appears insufficient to explain the relative resistance of zebrafish to dietary AFB1 hepatocarcinogenicity.
Troxel
C M
CM
Department of Food Science and Technology, Oregon State University, Corvallis 97331, USA.
Reddy
A P
AP
O'Neal
P E
PE
Hendricks
J D
JD
Bailey
G S
GS
D016604:Aflatoxin B1 / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001681:Biological Assay; D002273:Carcinogens / Q000378:metabolism* / Q000633:toxicity; D018736:DNA Adducts / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D007113:Immunity, Innate; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009374:Neoplasms, Experimental / Q000139:chemically induced / Q000503:physiopathology; D017686:Oncorhynchus mykiss / Q000378:metabolism; D013045:Species Specificity; D015027:Zebrafish / Q000378:metabolism*
D002273:Carcinogens; D018736:DNA Adducts; D016604:Aflatoxin B1