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Korean J Parasitol > Volume 31(4):1993 > Article

Original Article
Korean J Parasitol. 1993 Dec;31(4):353-361. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1993.31.4.353
Copyright © 1993 by The Korean Society for Parasitology
Localization and isozyme patterns of phosphatase in Fibricola seoulensis
H J Kim and C W Kim*
Department of Biology, Gyeongsang National University, Chinju 660-701, Korea.
Received August 17, 1993; Accepted September 20, 1993.

Abstract

The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acid phosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the non-substrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme band (62 kDa) of AcPase was recognized in metacercariae and 4 isozyme bands (95 kDa, 85 kDa, 73 kDa and 62 kDa) in adults.

Figures


Fig. 1
Histochemical localization of acid phosphatase in F. seoulensis treated with substrate, Naphthol AS-BI phosphate.

1~2, Treated at pH 2: 1. (A:adult) 40 ×, 2. (M: metacercaria) 40 ×, 3~4, Treated at pH 4:3. (A) 40 ×, 4. (M) 100 ×, 5~6, Treated at pH 5:5. (A) 40 ×, 6. (M) 100 ×. 7~8, Treated at pH 7:7. (A) 40 ×, 8. (M) 100 ×. OS: oral sucker, P: pharynx, VS: ventral sucker, IC: intestinal caecum, TO: tribocytic organ, TG: tribocytic gland



Fig. 2
Histochemical localization of acid phosphatase in F. seoulensis. 9~10, Treated without substrate: 7. (A:adult) 40 ×, 8. (M: metacercaria) 100 ×. 11~12, Treated with inhibitor (Oxalic acid): 9. (A) 40 ×, 10. (M) 100 ×.


Fig. 3
Histochemical localization of alkaline phosphatase in F. seoulensis treated with substrate, Naphthol AS-MX phosphate

13~14, Treated at pH 6 (A:40 ×, M: 40 ×), 15~16, Treated at pH 7 (A: 40 ×, M: 100 ×), 17~18, Treated at pH 8 (A: 40 ×, M 100 ×), 19~20, Treated at pH 9 (A:40 ×, M: 40 ×), IC: intestinal caecum, GA: genital anlagen TO: tribocytic organ EP: excretory pore



Fig. 4
Histochemical localization of alkaline phosphatase in F. seoulensis treated with substrate.

21~22, Treated at pH 7 (A: 40 ×, M: 100 ×), 23~24, Treated at pH 8 (A: 40 ×, M 100 ×), TG: tribocytic gland



Fig. 5
Protein fractions in extracts of F. seoulensis by non-denature PAGE and denature PAGE.

NA: non-denature adults, NM: non-denature metacercariae, DA: denature adult, DM: denature metacercariae, MW: molecular weight, Ma: marker protein, TD: tracking dye



Fig. 6
Isozyme patterns of acid phosphatase and general protein patterns in extracts of F. seoulensis by non-denature PAGE.

NA: non-denature adults, NM: non-denature metacercariae, MW: molecular weight, Ma: marker protein, 5: pH 5


Tables


Table 1
The activity and localization of acid phosphatase in adults and metacercariae of F. seoulensis in various pH valule


Table 2
The activity and localization of alkaline phosphatase in adult and metacerariae of F. seoulensis in various pH valule

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