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Megasomes as the targets of leucine methyl ester in Leishmania amazonensis amastigotes

Published online by Cambridge University Press:  06 April 2009

J.-C. Antoine ,
Colette Jouanne  and
Antoinette Ryter
J.-C. Antoine
Affiliation:
Unité d'Immunophysiologie cellulaire de l'Institut Pasteur et du C.N.R.S. (UA 1113), Département de Physiopathologie expérimentale
Colette Jouanne
Affiliation:
Unité d'Immunophysiologie cellulaire de l'Institut Pasteur et du C.N.R.S. (UA 1113), Département de Physiopathologie expérimentale
Antoinette Ryter
Affiliation:
Unité de Microscopie électronique de l'Institut Pasteur et du C.N.R.S. (UA 1148), Département de Biologie moléculaire Institut Pasteur, 25, rue du Dr. Roux, 75724 Paris Cédex 15, France
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Summary

Certain L-amino acid esters, such as L-leucine methyl ester (Leu-OMe), can kill intracellular and isolated Leishmania amazonensis amastigotes. Killing appears to involve ester trapping and hydrolysis within an acidified parasite compartment (M. Rabinovitch and S. C. Alfieri, 1987, Brazilian Journal of Medical and Biological Research 20, 665–74). We show here by acid phosphatase light microscopic cytochemistry and by ultrastructural morphometry that megasomes, lysosome-like amastigote organelles, are the putative parasite targets of Leu-OMe. This conclusion is supported by the following observations, (a) Control amastigotes displayed a string of electron-dense, acid phosphatase-positive megasomes mostly located in the cellular poles opposite the flagellar pockets. Incubation of the amastigotes with Leu-OMe resulted in concentration-dependent swelling and fusion of the organelles as well as decreased electron density of the internal contents. These changes, which preceded parasite disruption, were followed by the progressive loss of parasite viability and the release of acid phosphatase activity into the medium, (b) Incubation of the amastigotes with L-isoleucine methyl ester, a non-leishmanicidal compound, induced only moderate fusion of the megasomes. (c) Pre-incubation of the parasites with the proteinase inhibitors antipain and chymostatin, previously shown to confer protection from Leu-OMe toxicity, nearly completely prevented the morphological changes of megasomes. (d) Exposure of amastigotes to tryptophanamide (Trp-NH2), the leishmanicidal activity of which is not reduced by antipain and chymostatin, did not result in swelling and fusion of the megasomes. This last finding suggests that different mechanisms underlie the destruction of amastigotes by Trp-NH2 and Leu-OMe. Overall, the results are compatible with the hypothesis that Leu-OMe and other amino acid esters are trapped and hydrolysed within megasomes.

Keywords

Leishmania amazonensisamastigotemegasomeamino acid ester.
Type
Research Article
Information
Parasitology , Volume 99 , Issue 1 , August 1989 , pp. 1 - 9
Copyright
Copyright © Cambridge University Press 1989

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