In vitro characterization of cholinesterases in the earthworm Eisenia andrei.
Assessment of pollution impact in soil ecosystems has become a priority and interest has grown concerning the use of invertebrates as sentinel organisms. Inhibition of cholinesterase (ChE) activity has a great potential as a biomarker of pesticide exposure, and we evaluated the ChE kinetic parameters in the earthworm Eisenia andrei in the presence of acetylthiocholine (ASCh), proprionylthiocholine (PSCh) and butyrylthiocholine (BSCh). The highest ChE activity was found in the presence of ASCh and PSCh (42.45 and 49.82 nmol min(-1) mg protein(-1), respectively). BSCh was hydrolyzed at a rate of 4.04 nmol min(-1) mg protein(-1), but the time course did not reach a plateau under our experimental conditions. Km values were 0.142+/-0.006 and 0.183+/-0.053 mM for ASCh and PSCh, respectively. ASCh and PSCh hydrolysis were significantly inhibited by eserine (IC50 values were 1.44 x 10(-8) and 1.20 x 10(-8) M, respectively) and by carbaryl (IC50 values of 5.75 x 10(-9) and 4.79 x 10(-9) M). The presence of different ChEs in tissues from E. andrei was assessed by using selective inhibitors for AChE (BW284c51) and BChE (iso-OMPA). BW284c51 strongly reduced ASCh and PSCh hydrolysis and slightly affected that of BSCh, while iso-OMPA was without effect in all cases.
Caselli
Federico
F
University of Bologna, Interdepartment Centre for Research in Environmental Sciences (CIRSA), via S. Alberto 163, 48100 Ravenna, Italy.
Gastaldi
Laura
L
Gambi
Naimj
N
Fabbri
Elena
E
D000122:Acetylthiocholine / Q000378:metabolism; D000818:Animals; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D002092:Butyrylthiocholine / Q000378:metabolism; D012721:Carbaryl / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002802:Cholinesterases / Q000378:metabolism*; D007700:Kinetics; D009835:Oligochaeta / Q000201:enzymology*; D010830:Physostigmine / Q000494:pharmacology; D012989:Soil Pollutants / Q000032:analysis; D013379:Substrate Specificity; D013770:Tetraisopropylpyrophosphamide / Q000494:pharmacology; D013860:Thiocholine / Q000031:analogs & derivatives / Q000378:metabolism
D002800:Cholinesterase Inhibitors; D012989:Soil Pollutants; C009063:propionylthiocholine; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; D000122:Acetylthiocholine; D002092:Butyrylthiocholine; D013770:Tetraisopropylpyrophosphamide; D013860:Thiocholine; D010830:Physostigmine; D002802:Cholinesterases; D012721:Carbaryl