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Interplay between lysosomal, mitochondrial and death receptor pathways during manganese-induced apoptosis in glial cells

  • Inorganic Compounds
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Abstract

Manganese (Mn) is an essential trace metal which plays a critical role in brain physiology by acting as a cofactor for several enzymes. However, upon overexposure, Mn preferentially accumulates within the basal ganglia leading to the development of a Parkinsonism known as Manganism. Data from our group have proved that Mn induces oxidative stress-mediated apoptosis in astrocytoma C6 cells. In the present study we described how cathepsins impact on different steps of each apoptotic cascade. Evidence obtained demonstrated that Mn generates lysosomal membrane permeabilization (LMP) and cathepsin release. Both cathepsins B (Ca-074 Me) and D (Pepstatin A) inhibitors as well as Bafilomycin A1 prevented caspases-3, -7, -8 and -9 activation, FasL upregulation, Bid cleavage, Δφm disruption and cytochrome c release. Results from in vivo studies showed that intrastriatal Mn injection increased cathepsin D levels from corpus striatum and substantia nigra pars compacta. Our results point to LMP and lysosomal cathepsins as key mediators in the apoptotic process triggered by Mn. These findings highlight the relevance of targeting the lysosomal pathway for Manganism therapy.

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Acknowledgements

This work was supported by Grants from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET PIP 0356 and 0771). The authors thank Roberto Fernandez for his assistance in obtaining confocal microscopy images (IFIBYNE-CONICET, Argentina). R.M.G. and S.P.A are supported by CONICET scholarships. A.A., F.S. and M.L.K. are research members at CONICET.

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Authors and Affiliations

  1. CONICET- Universidad de Buenos Aires, Instituto de Química Biológica Ciencias Exactas y Naturales (IQUIBICEN), Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Laboratorio de Disfunción Celular en Enfermedades Neurodegenerativas y Nanomedicina, Buenos Aires, Argentina

    R. M. Gorojod, A. Alaimo, S. Porte Alcon & M. L. Kotler

  2. CONICET- Universidad de Buenos Aires, Instituto de Biologia y Medicina Experimental (IBYME), Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Laboratorio de Neurobiologia, Buenos Aires, Argentina

    F. Saravia

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  1. R. M. Gorojod
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Correspondence to M. L. Kotler.

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204_2017_1936_MOESM1_ESM.tif

Figure S1. Mn generates an increase in CatD expression levels. Cell cultures were exposed to Mn for 24h. Total cell lysates were run on SDS-PAGE. Blots were probed with anti-CatD and anti-β-Actin as a loading control (TIF 40 KB)

204_2017_1936_MOESM2_ESM.tif

Figure S2. CatD levels in the globus pallidus and the brain cortex after striatal Mn injection. Animals were injected into the striatum with 1µmol MnCl2 (left) and with saline as control (right). Rats were sacrificed 7 days post injection. Coronal slices containing the globus pallidus and the brain cortex were immunolabeled to detect CatD using DAB as the chromogen. Scale bar: 100µm (TIF 8490 KB)

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Gorojod, R.M., Alaimo, A., Porte Alcon, S. et al. Interplay between lysosomal, mitochondrial and death receptor pathways during manganese-induced apoptosis in glial cells. Arch Toxicol 91, 3065–3078 (2017). https://doi.org/10.1007/s00204-017-1936-7

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  • DOI: https://doi.org/10.1007/s00204-017-1936-7

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