Functional visualization of the excretory system of adult Schistosoma mansoni by the fluorescent marker resorufin

Excretion of metabolic wastes as well as xenobiotics is a major concern of all living organisms, and the Platyhelminthes including Schistosoma mansoni possess the protonephridial excretory system for their survival. Except for some ultrastructural and biochemical information, little is known about the protonephridium of platyhelminths due to a lack of established techniques for exploring the excretory activity. This study describes a new technique to assess the excretory activity of S. mansoni by use of the fluorescent marker resorufin, which is a potential substrate of the drug efflux pump, P-glycoprotein. After simple diffusion into the schistosome body, fluorescent resorufin was concentrated in the excretory tubules by an energy-dependent mechanism and excreted via the nephridiopore. The present technique of labelling functionally the excretory system was applicable to adult worms, but not schistosomula or cercariae. A variety of modulators known to interfere with mammalian P-glycoprotein function perturbed resorufin excretion from male adult schistosomes, including cyclosporin A, Ro11-2933, verapamil, or nifedipine. This technique of labelling the excretory system with fluorescent resorufin has enabled us to study aspects of the physiological function, hitherto unknown, of the protonephridial system of S. mansoni.