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- Title
Characterisation of cysteine proteinase activities in the digestive tract of juvenile Paragnathia formica isopods, ectoparasites of estuarine fish.
- Authors
Manship, Brigitte M.; Walker, Anthony J.; Jones, Lucy A.; Davies, Angela J.
- Abstract
Juveniles of Paragnathia formica Hesse (Isopoda; Gnathiidae) are haematophagous ectoparasites, feeding on fish blood which supplies the nutrients for their development through growth and moulting, and the subsequent survival and reproduction of the free-living adults. Little is known of the mechanisms of digestion in juvenile gnathiids, and biochemical studies on the digestive tract of these interesting estuarine isopods have not been undertaken previously. Here, functionally active cathepsin-like cysteine proteinases are identified in the digestive system of juvenile praniza (fed) and zuphea (unfed) forms. The physiological pH of the digestive tract and the optimum proteolytic activities detected in praniza 3 homogenates using the cathepsin B/L, cathepsin B, and cathepsin H fluorogenic substrates, N-Carbobenzoxy-Phe-Arg-4-methoxy-2-naphthylamine ( Z-phe-arg-MNA), N-Carbobenzoxy-Arg-Arg-4-methoxy-2-naphthylamine ( Z-arg-arg-MNA) and Arg-4-methoxy-2-naphthylamine ( H-arg-MNA), respectively, are in the acidic range (pH 5.8–6.7). Inhibition profiles against Z-phe-arg-MNA and Z-arg-arg-MNA using the cathepsin B inactivator urea, and cysteine proteinase inhibitors, support the presence of cathepsin L- and B-like enzymes. These proteolytic activities are 10–50 times higher in homogenates of praniza 3 compared with zuphea 3 forms. Histochemistry of praniza 3 sections reveals that the predominant enzyme activity towards Z-phe-arg-MNA is limited to the digestive glands during early and mid stages of digestion; later, this activity appears in the lining of the anterior hindgut. Moreover, activity towards Z-arg-arg-MNA is generally restricted to the digestive glands, and only occasionally present in the anterior hindgut. In conclusion, the digestive glands are the main site of cathepsin-like cysteine proteinase activities in P. formica juveniles; these enzymes appear to be important to the digestion of host fish blood enabling development through to the free-living adult.
- Subjects
CYSTEINE proteinases; ALIMENTARY canal; ESTUARINE fishes; ESTUARINE animals; FORMICA (Insects); BLOODSUCKING insects; HISTOCHEMISTRY; PARASITIC diseases; PROTEOLYTIC enzymes
- Publication
Marine Biology, 2008, Vol 153, Issue 3, p473
- ISSN
0025-3162
- Publication type
Article
- DOI
10.1007/s00227-007-0834-6