PERMEATION OF LYSOSOMAL MEMBRANES IN THE COURSE OF PHOTOSENSITIZATION WITH METHYLENE BLUE AND HEMATOPORPHYRIN: STUDY BY CELLULAR MICROSPECTROFLUOROMETRY |
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| Authors: | R Santus † C Kohen E Kohen ‡ J P Reyftmann † P Morliere § L Dubertret § P M Tocci &#; |
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| Institution: | *Papanicolaou Cancer Research Institute, 1155 N.W. 14th Street, P.O. Box 016188, Miami, Florida 33101, USA;†Laboratoire de Physico-Chimie de l'Adaptation Biologique, Equipe de Recherche Associée au CNRS No 951, Museum National d'Histoire Naturelle, 43 rue Cuvier, 75231, Paris Cedex 05, France;‡Also department of Biology, University of Miami, Coral Gables, Florida USA, Affiliate Faculty, Department of Pathology, School of Medicine, University of Miami, Miami, Florida USA.;§Hôpital Henri Mondor, Service de Dermatologie, 94010 Creteil, France, USA.;‖Department of Pediatrics, School of Medicine, University of Miami, Miami, Florida, USA |
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| Abstract: | Abstract— The photodynamically-induced liberation of lysosomal enzymes using ß-galactosidase as marker for the lysosomal enzymes has been studied by microspectrofluorometry on mouse L cells. Similar studies have been carried out using N-acetyl-ß-D-glucosaminidase as marker for the lysosomal enzymes of human fibroblasts. The high sensitivity of the fluorescence detection makes it possible to use 4-methylumbelliferyl substrates for the enzymes contained in a single cell. Methylene blue and hematoporphyrin readily incorporate into both cells and upon excitation, sensitize lysosomal membrane damages, leading to enzyme release accompanying strong morphological changes. |
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