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The nervous system of Fibricola seoulensis by acetylcholinesterase histochemistry
E W Cheon and C H Kim*
Department of Biology, Gyeongsang National University, Chinju 660-701, Korea.
Received August 21, 1993; Accepted September 20, 1993.
Abstract
F. seoulensis were obtained from artificially infected albino rats at 3, 4, 5, 6, 7 days after infection. The worms and metacercariae were washed in physiological saline solution, and fixed with 10% neutral formalin. The acetylcholinesterase (AchE) stained by enzyme histochemistry using acetylthiocholine iodide as substrate. Eserine, iso-OMPA and BW284C51 were used as inhibitors of AchE. The nervous system consists of three pairs longitudinal nerve trunks interconnected with excretory plexus in posterior half, and pharynx and oral sucker in anterior half of metacercariae and adults. The longitudinal nerve trunks are interconnected with transverse commissures and numerous circular commissures. Considerable numbers of circular commissures are interconnected with longitudinal nerve trunks lying on the surface of the worms. At each stage of juvenile worms, AchE and nonspecific cholinesterase activities were observed in the oral sucker, ventral sucker, pharynx and nerve system. Isozymes of AchE in F. seoulensis were separated into the two bands, 69 kDa and 132 kDa. The major band was 69 kDa.
Figures
Figs. 1-6 Fig. 1. Histochemical localization of acetylcholinesterase on metacercariae. Localization of AchE in metacercarae treated with substrate (Acetylthiocholine iodide). ×100. Fig. 2. Metacercariae treated with inhibitor, 10-4 M eserine for 1 hour at room temperature. ×100. Fig. 3. Histochemical localization of acetylcholinesterase of F. seoulensis developed in host for 6 days after infection. Localization of AchE in F. seoulensis treated with substrate (Acetylthiocholine iodide). ×40. Fig. 4.F. seoulensis developed in host for 6 days after infection treated with inhibitor, 10-4 M eserine for 1 hour at room temperature. ×40. Fig. 5. Histochemical localization of acetylcholinesterase on adult (7 days). Localization of AchE in F. seoulensis treated with substrate (Acetylthiocholine iodide). ×40. Fig. 6.F. seoulensis developed in host for 7 days after infection treated with inhibitor, 10-4 M eserine for 2 hour at room temperature. ×40.
Figs. 7-14 Fig. 7. Metacercariae treated with inhibitor, 10-4 M iso-OMPA for 2 hour at room temperature. ×100. Fig. 8. Metacercariae treated with inhibitor, 10-4 M BW284C51 for 2 hour at room temperature. ×100. Fig. 9. Histochemical localization of acetylcholinesterase on F. seoulensis developed in host for 3 days after infection. Localization of AchE in F. seoulensis treated with substrate (Acetylthiocholine iodide). ×40. Fig. 10.F. seoulensis developed in host for 3 days after infection treated with inhibitor, 10-4 M eserine for 2 hour at room temperature. ×40. Fig. 11.F. seoulensis developed in host for 3 days after infection treated with inhibitor, 10-4 M iso-OMPA for 2 hour at room temperature. ×40. Fig. 12.F. seoulensis developed in host for 3 days after infection treated with inhibitor, 10-4 M BW284C51 for 2 hour at room temperature. ×40. Fig. 13. Histochemical localization of acetylcholinesterase on F. seoulensis developed in host for 4 days after infection. Localization of AchE in F. seoulensis treated with substrate (Acetylthiocholine iodide). ×40. Fig. 14.F. seoulensis developed in host for 4 days after infection treated with inhibitor, 10-4 M eserine for 2 hour at room temperature. ×40. Os: oral sucker, P: pharynx, Vs: ventral sucker, Nm: neural mass, Lt: Longitudinal nerve trunk, Mlt: main longitudinal nerve trunk, Cc: circular commissure, Ep: excretory plexus, T: tribocytic organ.
Figs. 15-16 Histochemical localization of acetylcholinesterase on F. seoulensis developed in host for 6 days after infection. Localization of AchE in anterior and posterior portion of developmental adult treated with substrate (Acetylthiocholine iodide). ×200. Os: oral sucker, P: pharynx, Vs: ventral sucker, Nm: neural mass, Lt: Longitudinal nerve trunk, Mlt: main longitudinal nerve trunk, Cc: circular commissure, Ep: excretory plexus, T: tribocytic organ.
Fig. 17 Isozyme patterns of acetylcholinesterase of extracts of F. seoulensis by nondenature PAGE. FE: Treated inhibitor (eserine 10-4 M) in extracts of F. seoulensis, FB: Treated inhibitor (BW284C51 10-4 M) in extracts of F. seoulensis. F: Extracts of F. seoulensis treated with substrate (Acetylthiocholine iodide). MW: marker protein, Co: comassie blue staining, TD: tracking dye
Tables
Table 1 The histochemical localization and relative intensity of acetylcholinesterase activity on developing F. seoulensis in host
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