A fluorescent dye monochlorobimane (MCB) that binds glutathione
(GSH) was
used as a tool for measuring the
concentration of GSH in skin and mechanically-transformed schistosomula.
The
specificity of MCB binding to GSH was
confirmed by thin layer chromatography (TLC) and high performance liquid
chromatography (HPLC). The MCB
binding to GSH is an energy-dependent process since no labelling could
be seen at
low temperature. When 24-h-old
schistosomula were depleted of GSH by buthionine sulfoximine (a specific
inhibitor
of GSH synthesis) for 18 h, a
significant decrease (P<0·001) in fluorescence was observed.
PZQ
treatment of the schistosomula after first labelling the
parasites with MCB did not greatly affect MCB binding to GSH. However,
when the
24-h-old schistosomula were first
PZQ treated and afterwards labelled with MCB, the pattern of labelling
was
identical to that of those of the non-labelled
parasites. When 24-h-old schistosomula were first PZQ treated, washed and
labelled in the presence of 1 mM GSH, the
level of fluorescence was recovered. These results suggest that PZQ depletes
GSH from schistosomula, and may render
them susceptible to the host's immune system.