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Studies on the effect of aminochrome in cytoskeleton protein network and its role in protein degradation systems Irmgard Paris1,2.

Mnica Villa2, Gloria Garrido2, Karina Corts 1,2, Patricia Muoz2, Andrea Briceo2, Maricella Delgado1,2, Macarena Tapia 1,2, Mario Piones1 , Alejandra Caldern1, Constanza Toloza1, Mara Francisca Galleguillos1, Martha Naranjo1, Jennifer Encina1, Sandro Huenchuguala2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan Segura-Aguilar2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Parkinson disease (PD) is a devastating disorder that affect nervous system which is characterized by the loss of dopaminergic neurons in the (SNpc) substantia nigra pars compact. Despite intense worldwide research, the exact mechanisms involved in the pathogenesis of PD are unclear. Increasing evidence suggests PD is associated with the formation of intracytoplasmic protein aggregates (Lewy-body inclusions) in neurons of the SNpc and other brain areas. Several lines of evidence suggest autophagic dysregulation and dysfunction of the aggresome pathway may be responsible for accumulation of abnormal proteins or damaged organelles in this disease. Recently, we reported that aminochrome induces disruption of cytoskeleton by inducing aggregations of - and -tubulin. The dynein/dynactin complex has been involved in autophagy and lysosomal trafficking participating in the clearance of protein aggregates. In addition, the transport of autophagosome and aggresomes towards lysosomes is mediated by microtubules in association with dynein/dynactin complex. The present investigation tested the hypothesis that aminochrome affect the dynein/dynactin complex inducing the dysfunction of protein degradation systems. In this work, we have used a SH-SY5Y human neuroblastoma cell line. The results showed that significantly cell death of differentiated SH-SY5Y cells was observed when the cells were incubated with aminochrome. Under these conditions we observed protein expression changes and cytoskeleton protein aggregation. These results suggest that aminochrome may play a role in the dysfunction of protein degradation systems mediated by alterations in cytoskeleton protein network how is observed in PD. Supported by Fondecyt 1120337 and Project University Santo Toms N000012858.

Studies on the effect of copper in the disruption of cytoskeleton by inducing protein aggregations Mario Piones1, Martha Naranjo1, Mnica Villa2, Gloria Garrido2, Karina Corts 1,2, Patricia Muoz2, Sandro Huenchuguala2, Maricella Delgado 1,2, Macarena Tapia 1,2, Andrea Briceo2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Marianela Arvalo3, Juan Segura-Aguilar2, Irmgard Paris 1,2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. 3 Department of Medic Technology, Universidad Santo Toms, Via del Mar, Chile. Copper is an essential trace element playing an important role in cell physiology of our body. However, under certain circumstances this metal could be toxic. Role of copper in neurodegenerative disease as Parkinsons disease is suggested by the increased incidence of Parkinsonism in subjects exposed to copper who work smelting industry in Chile. Also, Parkinsonism is one of the major neurological symptoms in Wilson disease, a copper deposition disorder. Previous results suggested that copper neurotoxicity was dependent on the formation of Cu-dopamine complexes with concomitant dopamine oxidation to aminochrome and also on one-electron reduction of aminochrome. Recently, we reported that Cu-dopamine complex induces autophagy and aminochrome induces disruption of cytoskeleton by inducing aggregations of - and -tubulin, suggesting that aminochrome may play a role in the dysfunction of protein degradation systems observed in Parkinsons disease. It was described that HDAC6 and microtubules are required for autophagic degradation of aggregated protein. Likewise, the formation of aggresomes is also involved in eliminating misfolded protein. The present investigation tested the hypothesis that copper toxicity is related with the formation of cytoskeleton proteins aggregated that participated in the protein elimination pathway such as -III tubulin, gamma tubulin and histone deacetylase 6 (HDAC6). These proteins participate in both microtubules formation and stabilization and also protein degradation pathways. In this work, we have used a SH-SY5Y human neuroblastoma cell line. Our results showed a moderate cell death in presence of dopamine or copper alone without formation protein aggregates. By contrast, Cudopamine complex induces significantly cell death in differentiated SH-SY5Y cells with the formation -III tubulin, gamma tubulin and HDAC6 aggregates. These results suggest that Cu-dopamine complex leads disruption of cytoskeleton by inducing protein aggregations, suggesting that Cu-dopamine complex may generate the dysfunction of protein degradation systems how observed in Parkinsons disease. Supported by Fondecyt 1120337 and Project University Santo Toms N000012858.

Studies on the effect of iron in the disruption of cytoskeleton by inducing protein aggregations Jennifer Encina1, Alejandra Caldern1, Mara Francisca Galleguillos1, Mnica Villa2, Gloria Garrido2, Karina Corts 1,2, Patricia Muoz2, Sandro Huenchuguala2, Maricella Delgado 1,2, Macarena Tapia 1,2, Andrea Briceo2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan Segura-Aguilar2, Irmgard Paris 1,2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Iron is essential trace element for life; however under certain circumstances this metal could be toxic. The role of iron in the pathophysiology of Parkinsons disease is supported by the finding that iron accumulates in the brain regions affected by this disease. Previous results suggested that iron neurotoxicity was dependent on the formation of Fe-dopamine complexes with concomitant dopamine oxidation to aminochrome and also on one-electron reduction of aminochrome. Recently, we reported that aminochrome induces disruption of cytoskeleton by inducing aggregations of - and -tubulin, suggesting that aminochrome may play a role in the vesicles transport systems like observed in Parkinsons disease. It was described that myosinVa, microfilaments and microtubules are required for vesicles transport into the axon end. The present investigation tested the hypothesis that iron toxicity is related with the formation of cytoskeleton proteins aggregated that participated in the vesicles transport systems such as tubulin, actin and myosinVa. In this work, we have used a SH-SY5Y human neuroblastoma cell line. Our results showed a moderate cell death in presence of dopamine or iron alone without formation protein aggregates. By contrast, Fe-dopamine complex induces significantly cell death in differentiated SH-SY5Y cells with the formation acetylated tubulin, and myosinVa aggregates. These results suggest that Fedopamine complex leads disruption of cytoskeleton by inducing protein aggregations, suggesting that Fe-dopamine complex may generate the dysfunction in the vesicles release and transport how observed in Parkinsons disease. Supported by Fondecyt 1120337.

Studies on the effect of manganese in the cytoskeleton network Constanza Toloza1, Jennifer Encina1, Mnica Villa2, Gloria Garrido2, Karina Corts 1,2, Patricia Muoz2, Sandro Huenchuguala2, Maricella Delgado 1,2, Macarena Tapia 1,2, Andrea Briceo2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan Segura-Aguilar2, Irmgard Paris 1,2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Manganese is an essential element for brain development; however under certain circumstances this metal could be toxic. Manganese exposure has been associated with the development of Parkinsonism. Previous results suggested that manganese neurotoxicity was dependent on the dopamine oxidation to aminochrome and also on one-electron reduction of aminochrome. Recently, we reported that aminochrome induces disruption of cytoskeleton by inducing aggregations of actin and - and tubulin. Microfilaments and microtubules are required not only for vesicles transport into the axon end but also for the clearance of protein aggregates. The present investigation tested the hypothesis that manganese toxicity is related with the formation of cytoskeleton proteins aggregated that participated in the vesicles transport systems and autophagy/aggresome pathways such as tubulin, actin and gamma tubulin. In this work, we have used a SH-SY5Y human neuroblastoma cell line. Our results showed a moderate cell death in presence of dopamine or manganese alone without formation protein aggregates. By contrast, manganese in presence of dopamine induces significantly cell death in differentiated SH-SY5Y cells with the formation gamma tubulin, -III tubulin and actin aggregates. These results suggest that manganese neurotoxicity dependent on the dopamine oxidation to aminochrome leads disruption of cytoskeleton by inducing protein aggregations which could generate the dysfunction in the vesicles transport and clearance of protein aggregates how observed in Parkinsons disease. Supported by Fondecyt 1120337.

Studies on the effect of aminochrome in cytoskeleton protein and its role in protein degradation systems Gloria Garrido2, Mnica Villa2, Andrea Briceo2, Patricia Muoz2, Maricella Delgado1,2, Macarena Tapia1,2, Karina Corts 1,2, Sandro Huenchuguala2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan Segura-Aguilar2, Irmgard Paris1,2
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Parkinson disease (PD) is neurodegenerative disorder that affects nervous system which is characterized by the loss of dopaminergic neurons in the (SNpc) substantia nigra pars compact. Currently, the exact mechanisms involved in the pathogenesis of PD are unclear. Increasing evidence suggests PD is associated with the formation of intracytoplasmic protein aggregates in neurons of the SNpc and other brain areas. Several lines of evidence suggest autophagic dysregulation and dysfunction of the aggresome pathway may be responsible for accumulation of abnormal proteins or damaged organelles in this disease. Recently, we reported that aminochrome induces disruption of cytoskeleton by inducing aggregations of - and -tubulin. The dynein/dynactin complex has been involved in autophagy/aggresome pathway participating in the clearance of protein aggregates. In addition, the transport of autophagosome and aggresomes towards lysosomes is mediated by microtubules in association with dynein/dynactin complex. The present investigation tested the hypothesis that aminochrome affect the dynein/dynactin complex inducing the dysfunction of protein degradation systems. In this work, we have used a SH-SY5Y human neuroblastoma cell line. The results showed that significantly cell death of differentiated SH-SY5Y cells was observed when the cells were incubated with aminochrome. Under these conditions we observed protein expression changes related with autophagy/aggresome pathway. These results suggest that aminochrome may play a role in the dysfunction of protein degradation systems mediated by alterations in cytoskeleton protein network how is observed in PD. Supported by Fondecyt 1120337.

Studies on the effect of aminochrome in the microtubules network and its association with motor proteins Andrea Briceo2, Gloria Garrido2, Mnica Villa2, Patricia Muoz2, Maricella Delgado1,2, Macarena Tapia1,2, Karina Corts1,2, Sandro Huenchuguala2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan Segura-Aguilar2, Irmgard Paris1,2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Parkinson disease (PD) is one of most neurodegenerative disorder that affects nervous system. This disease is characterized by the loss of dopaminergic neurons in the (SNpc) substantia nigra pars compact. Although, the exact mechanisms involved in the pathogenesis of PD are unclear, increasing evidence suggests PD is associated with the formation of intracytoplasmic protein aggregates in neurons of the SNpc and other brain areas. Several lines of evidence suggest dysfunction of the autophagic/aggresome pathway may be responsible for accumulation of abnormal proteins or damaged organelles in this disease. Recently, we reported that aminochrome induces disruption of cytoskeleton by inducing aggregations of - and -tubulin. The dynein/dynactin complex has been involved in autophagy/aggresome pathway participating in the clearance of protein aggregates. In addition, the transport of autophagosome and aggresomes towards lysosomes is mediated by microtubules in association with dynein/dynactin complex. The present investigation tested the hypothesis that aminochrome affect the microtubules polymerization and its interaction with dynein/dynactin complex. In this work, we have used a SH-SY5Y human neuroblastoma cell line. The results showed that aminochrome alter the ratio monomer/polymer tubulin affecting thus the association of microtubules with dynein/dynactin complex. These results suggest that aminochrome may play a role in the vesicles transport and protein degradation systems mediated by alterations in microtubules network how is observed in PD.

Supported by Fondecyt 1120337.

Studies on epigenetic alterations induced by aminochrome Karina Corts1,2, Patricia Muoz2, Mnica Villa2, Gloria Garrido2, Maricella Delgado1,2, Macarena Tapia1,2, Andrea Briceo2, Sandro Huenchuguala2, Carlos Cuevas2, Ulises Ahumada2, Catalina Melndez2, Sebastin Castillo2, Andrea Herrera2, Juan SeguraAguilar2, Irmgard Paris1,2.
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Department of Basic Sciences, University Santo Toms, Via del Mar, Chile. Programme of Molecular and Clinical Pharmacology, Faculty of Medicine, Santiago, Chile. Parkinson disease (PD) is one of most neurodegenerative disorder that affects nervous system. This disease is characterized by the loss of dopaminergic neurons in the (SNpc) substantia nigra pars compact. Although, the exact mechanisms involved in the pathogenesis of PD are unclear, increasing evidence suggests genetics and epigenetic factors are involved in this disorder. Diverse epigenetic mechanisms, such as DNA methylation, histone modifications and miRNA expression may play a key role in disease development. The present investigation tested the hypothesis that aminochrome induces aggregation of histone deacetylase (HDAC) affecting their histone deacetylase activity. In this work, we have used a SH-SY5Y human neuroblastoma cell line. The results showed that aminochrome induces significantly cell death of differentiated SHSY5Y cells; and decrease HDACs activity and expression. These results suggest that aminochrome may play a key role in the epigenetic regulation mechanisms in PD.

Supported by Fondecyt 1120337.