Hazardous effect of organophosphate compound, dichlorvos in transgenic Drosophila melanogaster (hsp70-lacZ): induction of hsp70, anti-oxidant enzymes and inhibition of acetylcholinesterase.
We tested a working hypothesis that stress genes and anti-oxidant enzyme machinery are induced by the organophosphate compound dichlorvos in a non-target organism. Third instar larvae of Drosophila melanogaster transgenic for hsp70 were exposed to 0.1 to 100.0 ppb dichlorvos and 5.0 mM CuSO(4) (an inducer of oxidative stress and stress genes) and hsp70, and activities of acetylcholinesterase (AchE), superoxide dismutase (SOD), catalase (CAT) and lipid peroxidation (LPO) product were measured. The study was further extended to examine tissue damage, if any, under such conditions. A concentration- and time-dependent increase in hsp70 and anti-oxidant enzymes was observed in the exposed organism as compared to control. A comparison of stress gene expression with SOD, CAT activities and LPO product under similar experimental conditions revealed that induction of hsp70 precedes the anti-oxidant enzyme activities in the exposed organism. Further, concomitant with a significant inhibition of AChE activity, significant induction of hsp70 was observed following chemical exposure. Mild tissue damage was observed in the larvae exposed to 10.0 ppb dichlorvos for 48 h when hsp70 expression reaches plateau. Dichlorvos at 0.1 ppb dietary concentration did not evoke significant hsp70 expression, anti-oxidant enzymes and LPO and AchE inhibition in the exposed organism, and thereby, was found to be non-hazardous to D. melanogaster. Conversely, 1.0 ppb of the test chemical stimulated a significant induction of hsp70 and anti-oxidant enzymes and significant inhibition of AchE; hence this concentration of test chemical was hazardous to the organism. The present study suggests that (a) both stress genes and anti-oxidant enzymes are stimulated as indices of cellular defense against xenobiotic hazard in D. melanogaster with hsp70 being proposed as first-tier bio-indicator of cellular hazard, (b) 0.1 ppb of the test chemical may be regarded as No Observed Adverse Effect Level (NOAEL), and 1.0 ppb dichlorvos as Low Observed Adverse Effect Level (LOAEL).
Gupta
Subash Chandra
SC
Embryotoxicology Section, Industrial Toxicology Research Center, Lucknow 226001, Uttar Pradesh, India.
Siddique
Hifzur Rahman
HR
Saxena
Daya Krishna
DK
Chowdhuri
Debapratim Kar
DK
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D000975:Antioxidants / Q000378:metabolism*; D002374:Catalase / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D019327:Copper Sulfate / Q000494:pharmacology; D004006:Dichlorvos / Q000494:pharmacology*; D004331:Drosophila melanogaster / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D004790:Enzyme Induction / Q000187:drug effects; D005724:Ganglia / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D018869:Heat-Shock Response / Q000502:physiology; D007150:Immunohistochemistry; D007814:Larva / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D015227:Lipid Peroxidation; D013482:Superoxide Dismutase / Q000378:metabolism; D014343:Trypan Blue
D000975:Antioxidants; D002800:Cholinesterase Inhibitors; D018840:HSP70 Heat-Shock Proteins; D004006:Dichlorvos; D002374:Catalase; D013482:Superoxide Dismutase; D000110:Acetylcholinesterase; D014343:Trypan Blue; D019327:Copper Sulfate