Fasciola hepatica: localisation of glutathione S-transferase isoenzymes in adult and juvenile liver fluke.
Four cDNA clones (GST-1, -7, -47, and -51) encoding isoenzymes of the detoxification enzyme glutathione S-transferase (GST) have previously been identified and characterised from Fasciola hepatica. In the present study, antisera were generated to synthetic peptides of regions unique to each of the four GST proteins predicted by the cDNAs. The antisera were characterised, and two were found to distinguish GST-1 from GST-7, GST-47, and GST-51 as a group. These two antisera were used to localise different GSTs in adult and newly excysted juvenile F. hepatica. The antiserum to GST-1 was specific and localised GST-1 to the parenchyma of adult fluke but not to the lamellae of the intestinal caeca. The antiserum to a GST-51 peptide, which cross-reacted with GST-7 and GST-47 but not GST-1, localised the other GSTs not only to the parenchyma but also to the intestinal lamellae of adult fluke. This appears to be the first evidence of tissue-specific expression of GST isoenzymes in trematodes. In contrast to adult fluke, immunolocalisation of the GSTs in juvenile F. hepatica revealed the binding of both the GST-1 and GST-51 antisera to the parenchymal cytoplasm, to cytoplasmic extensions of the parenchyma cells in the subtegumental area, as well as the excretory ducts. No labeling was observed in the intestinal epithelium of the juvenile fluke. These results demonstrate that adult F. hepatica, in contrast to juvenile flukes, contain a GST, which is not GST-1, associated with the lamellae of the gut and suggest that GSTs in adult fluke may play a role in the absorptive function of the adult gut.
Creaney
J
J
Victorian Institute of Animal Science, Department of Agriculture, Australia.
Wijffels
G L
GL
Sexton
J L
JL
Sandeman
R M
RM
Spithill
T W
TW
Parsons
J C
JC
D000375:Aging / Q000378:metabolism*; D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D005210:Fasciola hepatica / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D005982:Glutathione Transferase / Q000032:analysis / Q000378:metabolism*; D007106:Immune Sera; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity
D007106:Immune Sera; D007527:Isoenzymes; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D005982:Glutathione Transferase