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Manganese (Mn) overexposure is frequently associated with the development of a neurodegenerative disorder known as Manganism. The Mn-mediated generation of reactive oxygen species (ROS) promotes cellular damage, finally leading to apoptotic cell death in rat astrocytoma C6 cells. In this scenario, the autophagic pathway could play an important role in preventing cytotoxicity. In the present study, we found that Mn induced an increase in the amount and total volume of acidic vesicular organelles (AVOs), a process usually related to the activation of the autophagic pathway. Particularly, the generation of enlarged AVOs was a ROS- dependent event. In this report we demonstrated for the first time that Mn induces autophagy in glial cells. This conclusion emerged from the results obtained employing a battery of autophagy markers: a) the increase in LC3-II expression levels, b) the formation of autophagic vesicles labeled with monodansylcadaverine (MDC) or LC3 and, c) the increase in Beclin 1/ Bcl-2 and Beclin 1/ Bcl-XL ratio. Autophagy inhibition employing 3-MA and mAtg5K130R resulted in decreased cell viability indicating that this event plays a protective role in Mn- induced cell death. In addition, mitophagy was demonstrated by an increase in LC3 and TOM-20 colocalization. On the other hand, we proposed the occurrence of lysosomal membrane permeabilization (LMP) based in the fact that cathepsins B and D activities are essential for cell death. Both cathepsin B inhibitor (Ca-074 Me) or cathepsin D inhibitor (Pepstatin A) completely prevented Mn- induced cytotoxicity. In addition, low dose of Bafilomycin A1 showed a similar effect, a finding that adds evidence about the lysosomal role in Mn cytotoxicity. Finally, in vivo experiments demonstrated that Mn induces injury and alters LC3 expression levels in rat striatal astrocytes. In summary, our results demonstrated that autophagy is activated to counteract the harmful effect caused by Mn. These data is valuable to be considered in future research concerning Manganism therapies. © 2015 Published by Elsevier Inc.


Documento: Artículo
Título:The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions
Autor:Gorojod, R.M.; Alaimo, A.; Porte Alcon, S.; Pomilio, C.; Saravia, F.; Kotler, M.L.
Filiación:Departamento de Química Biológica, Universidad de Buenos Aires, Instituto de Química Biológica, Ciencias Exactas y Naturales (IQUIBICEN), Avda. Intendente Güiraldes 2 160Buenos Aires C1428EGA, Argentina
Departamento de Química Biológica, Universidad de Buenos Aires, Instituto de Biología y Medicina Experimental (IBYME)Buenos Aires, Argentina
Palabras clave:Acidic vesicular organelles; Autophagy; Cathepsin; Glial cells; Lysosomal cell death; Manganese; Mitophagy; Reactive oxygen species; bafilomycin A1; beclin 1; cathepsin B; cathepsin B inhibitor; dansylcadaverine; manganese; protein bcl 2; protein bcl x; LC3 protein, rat; manganese; microtubule associated protein; reactive oxygen metabolite; animal cell; Article; astrocyte; autophagy; cell death; cell permeabilization; cell viability; cytotoxicity; glia cell; lysosome; lysosome membrane; mitophagy; mouse; nonhuman; oxidative stress; priority journal; animal; apoptosis; astrocytoma; autophagy; biosynthesis; cell survival; drug effects; gene expression regulation; genetics; glia; lysosome; metabolism; pathology; rat; tumor cell line; Rattus; Animals; Apoptosis; Astrocytoma; Autophagy; Cell Line, Tumor; Cell Survival; Gene Expression Regulation; Lysosomes; Manganese; Metabolic Networks and Pathways; Microtubule-Associated Proteins; Neuroglia; Oxidative Stress; Rats; Reactive Oxygen Species
Página de inicio:237
Página de fin:251
Título revista:Free Radical Biology and Medicine
Título revista abreviado:Free Radic. Biol. Med.
CAS:bafilomycin A1, 88899-55-2; cathepsin B, 9047-22-7; dansylcadaverine, 10121-91-2; manganese, 16397-91-4, 7439-96-5; protein bcl 2, 219306-68-0; LC3 protein, rat; Manganese; Microtubule-Associated Proteins; Reactive Oxygen Species


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---------- APA ----------
Gorojod, R.M., Alaimo, A., Porte Alcon, S., Pomilio, C., Saravia, F. & Kotler, M.L. (2015) . The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions. Free Radical Biology and Medicine, 87, 237-251.
---------- CHICAGO ----------
Gorojod, R.M., Alaimo, A., Porte Alcon, S., Pomilio, C., Saravia, F., Kotler, M.L. "The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions" . Free Radical Biology and Medicine 87 (2015) : 237-251.
---------- MLA ----------
Gorojod, R.M., Alaimo, A., Porte Alcon, S., Pomilio, C., Saravia, F., Kotler, M.L. "The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions" . Free Radical Biology and Medicine, vol. 87, 2015, pp. 237-251.
---------- VANCOUVER ----------
Gorojod, R.M., Alaimo, A., Porte Alcon, S., Pomilio, C., Saravia, F., Kotler, M.L. The autophagic- lysosomal pathway determines the fate of glial cells under manganese- induced oxidative stress conditions. Free Radic. Biol. Med. 2015;87:237-251.