Journal of Environmental Biology

Gene expression of acetylcholinesterase in hybrid catfish (Clarias gariepinus X Clarias macrocephalus) exposed to chlorpyrifos and carbaryl

 

Chawanrat Somnuek¹, Chuta Boonphakdee², Voravit Cheevaporn*¹ and Keiichi Tanaka³

¹Graduate School of Environmental Science, Faculty of Science, Burapha University, Chonburi - 20131, Thailand

²Department of Biology, Faculty of Science, Burapha University, Chonburi - 20131, Thailand

³Faculty of Pharmacy, Osaka Ohtani University, Tondabayashi, Osaka - 584-8540, Japan

(Received:March 05, 2008 ; Revised received:May 10, 2008; Accepted: June 20,2008)

Abstract: After serving as a neurotransmitter, acetylcholine is hydrolyzed by acetylcholinesterase (AChE). Inhibition of AChE is considered to be a specific biomarker for exposure to organophosphate and carbamate insecticides. In this study, the AChE gene was isolated from hybrid catfish brain by RT-PCR methods using degenerate primers. The amplified fragment of 966 nucleotides generated by PCR was cloned, and sequence analysis showed 82% nucleotide identity with AChE of the Electrophorus electricus. Specific primers of the hybrid catfish AChE gene were then synthesized and used in the examination of AChE gene expression in brain tissue of hybrid catfish exposed to sublethal concentrations of chlorpyrifos (0.43, 4.3 and 43 ?M) and carbaryl (1.19, 11.9 and 119 ?M) for 24 hr. Real-time PCR was used to compare with the amplified 28S rRNA gene. AChE gene expression was significantly elevated 12.4 times in catfish exposed to 43 ?M chlorpyrifos in comparison to the control group (p<0.05). Carbaryl did not produce any significant change. These results indicate that AChE is more sensitive to administration of chlorpyrifos than carbaryl. The induction of the AChE gene indicates the possibility of using this gene as biomarker for detecting effects of organophosphate insecticides in hybrid catfish.

Key words: Acetylcholinesterase, Organophosphate, Carbamate, Chlorpyrifos, Carbaryl, Hybrid catfish, Real-time PCR

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