Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by neuromuscular connectivity decline followed by motoneuron loss. Altered proteostasis is suggested as a transversal pathogenic mechanism, notably involving dysfunction at the level of the endoplasmic reticulum (...

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Main Authors: Martin Sepulveda, Francisca MartinezTraub, Patricia Ojeda, Jessica Mella, Jorge Ojeda, Cristina Pinto, Rodrigo Diaz, Pablo Rozas, Claudia Sepulveda, Bredford Kerr, Vania Morales, Mirva Saaranen, Lloyd Ruddock, Danilo B. Medinas, Juan Pablo Henriquez, Claudio Hetz
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Language:English
Published: Elsevier 2025-10-01
Series:Neurobiology of Disease
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Online Access:http://www.sciencedirect.com/science/article/pii/S096999612500261X
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author Martin Sepulveda
Francisca MartinezTraub
Patricia Ojeda
Jessica Mella
Jorge Ojeda
Cristina Pinto
Rodrigo Diaz
Pablo Rozas
Claudia Sepulveda
Bredford Kerr
Vania Morales
Mirva Saaranen
Lloyd Ruddock
Danilo B. Medinas
Juan Pablo Henriquez
Claudio Hetz
author_facet Martin Sepulveda
Francisca MartinezTraub
Patricia Ojeda
Jessica Mella
Jorge Ojeda
Cristina Pinto
Rodrigo Diaz
Pablo Rozas
Claudia Sepulveda
Bredford Kerr
Vania Morales
Mirva Saaranen
Lloyd Ruddock
Danilo B. Medinas
Juan Pablo Henriquez
Claudio Hetz
author_sort Martin Sepulveda
collection DOAJ
description Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by neuromuscular connectivity decline followed by motoneuron loss. Altered proteostasis is suggested as a transversal pathogenic mechanism, notably involving dysfunction at the level of the endoplasmic reticulum (ER). Protein disulfide isomerases (PDIs) are key enzymes that catalyze protein folding and disulfide bond formation in the ER. Importantly, PDIs function is disrupted in ALS. We previously identified mutations in the gene encoding PDIA3 (also known as Grp58 or ERp57) as risk factors for ALS, which were associated with altered neuromuscular junction (NMJ) organization when expressed in zebrafish, a phenotype recapitulated in PDIA3-null mice. Here, we generated a transgenic mouse line overexpressing the ALS-linked PDIA3 variant D217N and performed a comprehensive characterization of ALS-like features. The transgenic line exhibited moderate overexpression of mutant PDIA3D217N, which led to morphological alterations at the NMJ resembling those observed in ALS models and patients, along with abnormal distribution of oxidative and glycolytic muscle fibers. However, mutant PDIA3D217N expression did not result in motor impairment, coordination deficits, or motoneuron loss. At the molecular level, we observed reduced expression of SV2 in the spinal cord, an important synaptic protein involved in NMJ function. Our findings further support the involvement of PDIA3 dysfunction as a risk factor in the emergence of early features of ALS.
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spelling doaj-art-b950470a460a43dfbcb829670c81e45d2025-08-20T03:41:43ZengElsevierNeurobiology of Disease1095-953X2025-10-0121410704510.1016/j.nbd.2025.107045Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junctionMartin Sepulveda0Francisca MartinezTraub1Patricia Ojeda2Jessica Mella3Jorge Ojeda4Cristina Pinto5Rodrigo Diaz6Pablo Rozas7Claudia Sepulveda8Bredford Kerr9Vania Morales10Mirva Saaranen11Lloyd Ruddock12Danilo B. Medinas13Juan Pablo Henriquez14Claudio Hetz15Institute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileCenter for Geroscience, Brain Health and Metabolism, Santiago, Chile; Neuromuscular Studies Lab (NeSt Lab), Instituto de Anatomía, Histología y Patología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, Chile; Department of Cell Biology, Faculty of Biological Sciences, Universidad de Concepción, Concepción, ChileDepartment of Cell Biology, Faculty of Biological Sciences, Universidad de Concepción, Concepción, ChileDepartment of Cell Biology, Faculty of Biological Sciences, Universidad de Concepción, Concepción, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, ChileCentro de Biología Celular y Biomedicina - CEBICEM, Facultad de Ciencias, Universidad San Sebastián, Chile; Centro de Estudios Científicos, Valdivia, ChileInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, Chile; Buck Institute for Research on Aging, Novato, CA, USAFaculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FinlandFaculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FinlandInstitute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, Chile; Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, BrazilNeuromuscular Studies Lab (NeSt Lab), Instituto de Anatomía, Histología y Patología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, Chile; Department of Cell Biology, Faculty of Biological Sciences, Universidad de Concepción, Concepción, Chile; Corresponding authors at: Institute of Biomedical Sciences, University of Chile, Independencia 1027, Santiago, Chile (CH) and Neuromuscular Studies Lab (NeSt Lab), Instituto de Anatomía, Histología y Patología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, Chile (JPH).Institute of Biomedical Sciences (ICBM), Faculty of Medicine, University of Chile, Santiago, Chile; Center for Geroscience, Brain Health and Metabolism, Santiago, Chile; Biomedical Neuroscience Institute (BNI) Corporation, University of Chile, Santiago, Chile; Buck Institute for Research on Aging, Novato, CA, USA; Corresponding authors at: Institute of Biomedical Sciences, University of Chile, Independencia 1027, Santiago, Chile (CH) and Neuromuscular Studies Lab (NeSt Lab), Instituto de Anatomía, Histología y Patología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, Chile (JPH).Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by neuromuscular connectivity decline followed by motoneuron loss. Altered proteostasis is suggested as a transversal pathogenic mechanism, notably involving dysfunction at the level of the endoplasmic reticulum (ER). Protein disulfide isomerases (PDIs) are key enzymes that catalyze protein folding and disulfide bond formation in the ER. Importantly, PDIs function is disrupted in ALS. We previously identified mutations in the gene encoding PDIA3 (also known as Grp58 or ERp57) as risk factors for ALS, which were associated with altered neuromuscular junction (NMJ) organization when expressed in zebrafish, a phenotype recapitulated in PDIA3-null mice. Here, we generated a transgenic mouse line overexpressing the ALS-linked PDIA3 variant D217N and performed a comprehensive characterization of ALS-like features. The transgenic line exhibited moderate overexpression of mutant PDIA3D217N, which led to morphological alterations at the NMJ resembling those observed in ALS models and patients, along with abnormal distribution of oxidative and glycolytic muscle fibers. However, mutant PDIA3D217N expression did not result in motor impairment, coordination deficits, or motoneuron loss. At the molecular level, we observed reduced expression of SV2 in the spinal cord, an important synaptic protein involved in NMJ function. Our findings further support the involvement of PDIA3 dysfunction as a risk factor in the emergence of early features of ALS.http://www.sciencedirect.com/science/article/pii/S096999612500261XAmyotrophic lateral sclerosisEndoplasmic reticulumProtein disulfide isomeraseProteostasisNeuromuscular junction
spellingShingle Martin Sepulveda
Francisca MartinezTraub
Patricia Ojeda
Jessica Mella
Jorge Ojeda
Cristina Pinto
Rodrigo Diaz
Pablo Rozas
Claudia Sepulveda
Bredford Kerr
Vania Morales
Mirva Saaranen
Lloyd Ruddock
Danilo B. Medinas
Juan Pablo Henriquez
Claudio Hetz
Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
Neurobiology of Disease
Amyotrophic lateral sclerosis
Endoplasmic reticulum
Protein disulfide isomerase
Proteostasis
Neuromuscular junction
title Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
title_full Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
title_fullStr Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
title_full_unstemmed Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
title_short Expression of amyotrophic lateral sclerosis associated protein disulfide isomerase A3 D217N variant recapitulates early morphological alterations at the neuromuscular junction
title_sort expression of amyotrophic lateral sclerosis associated protein disulfide isomerase a3 d217n variant recapitulates early morphological alterations at the neuromuscular junction
topic Amyotrophic lateral sclerosis
Endoplasmic reticulum
Protein disulfide isomerase
Proteostasis
Neuromuscular junction
url http://www.sciencedirect.com/science/article/pii/S096999612500261X
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