Reconstitution of Leishmania plasma membrane to understand the photodynamic effect
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Biophysical Reviews
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INTRODUCTION
Leishmaniasis is an important neglected disease. Photodynamic therapy
(PDT) has been used to fight cutaneous leishmaniasis showing good
results. However, PDT mechanisms in Leishmania parasites are not yet
completely clarified.
OBJECTIVES
In this work, our objective was to develop a protocol to produce giant
plasma membrane vesicles (GPMVs) from Leishmania amazonensis
promastigotes to understand the mechanisms of action of methylene blue
(MB)-mediated PDT on the cell membrane of parasites.
MATERIALS AND METHODS
For membrane extraction, several techniques were tested. The osmotic
shock was the technique that presented the best yield and effectiveness.
Phosphate and protein measurements were performed to confirm membrane
extraction. For the growth of GPMVs, the best technique was
electroforming using different frequencies and voltages in 4 cycles.
Reconstituted GPMVs were observed by phase-contrast light microscopy.
Subsequently, PDT was applied to GPMVs dispersed in an aqueous
solution containing 50 μM MB and we verified the changes in permeability
before and after exposure to light. The same processwas applied to
giant unilamellar vesicles (GUVs) with lipid compositions similar to the
parasite membrane.
DISCUSSION AND RESULTS
The electroforming technique with the protocol developed in this work
made it possible to obtain GPMVs froma promastigote membrane isolate
of L. amazonensis. The membrane isolation technique was effective to
extract the parasite's membrane while preserving lipids and proteins. In
GUVs we observe an increase in the area during PDT in different compositions
and loss of contrast. The GPMVs showed a loss of contrast as
well as the GUVs but did not show an increase in area.
CONCLUSION
This factor could be explained by the high degree of complexity of the
membrane, which contains membrane proteins in addition to containing
lipids.
Como referenciar
SOUZA, MARESSA D.F. de; ITRI, ROSANGELA; RIBEIRO, MARTHA S. Reconstitution of Leishmania plasma membrane to understand the photodynamic effect. Biophysical Reviews, v. 13, n. 6, p. 1384-1384, 2021. DOI: 10.1007/s12551-021-00845-2. Disponível em: http://repositorio.ipen.br/handle/123456789/32887. Acesso em: 30 Dec 2025.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.