Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection

Objective: Metabolic disruption is a central feature to many neurodegenerative diseases. Despite this, many gaps exist in our understanding of how these perturbations link to the mechanisms of neural disease. In this study, we sought to understand how genetically-controlled, cell-specific loss of py...

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Main Authors: Thomas J. Waller, Catherine A. Collins, Monica Dus
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:Molecular Metabolism
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Online Access:http://www.sciencedirect.com/science/article/pii/S2212877825000948
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author Thomas J. Waller
Catherine A. Collins
Monica Dus
author_facet Thomas J. Waller
Catherine A. Collins
Monica Dus
author_sort Thomas J. Waller
collection DOAJ
description Objective: Metabolic disruption is a central feature to many neurodegenerative diseases. Despite this, many gaps exist in our understanding of how these perturbations link to the mechanisms of neural disease. In this study, we sought to understand how genetically-controlled, cell-specific loss of pyruvate kinase (PyK) impacts motor neuron synaptic integrity and how the canonical neurodegenerative proteins DLK and SARM1 respond to this break in homeostasis. Methods: This study made use of the genetically-tractable Drosophila melanogaster to cell-specifically express proteins (via the GAL4/UAS binary system), knockdown gene transcripts (via RNA interference), and knockout gene loci (via guide RNA-directed Cas9). Synaptic and axonal degeneration were measured through immunohistochemistry, microscopy, and blinded scoring of fly larvae at both early and later 3rd instar stages to test for progressive phenotypes. Nervous system injury through a physical nerve crush assay was used to assay functional outcomes of protective stress responses. Results: We found that knockdown or knockout of PyK results in progressive axonal and synaptic degeneration, dependent on signaling through DLK and SARM1. This degeneration is preceded by nuclear transcriptional activation by DLK and the downstream AP-1 transcription factor Fos. We also found evidence of a neuroprotective response through injury of PyK-deficient axons (before progressive degeneration has occurred), which results in delayed Wallerian degeneration. This delay shows dependence on DLK and Fos, and coincides with reduced axonal localization of SARM1 whose overexpression fully restores degeneration speed. Conclusions: These data support a rheostat model of DLK signaling that both promotes and inhibits axon degeneration in response to metabolic disruption. This rheostat likely converges on regulation of SARM1, which is required for the progressive synapse loss following PyK, but also abolishes the protective delay in injury-induced Wallerian degeneration when overexpressed. Overall, we conclude that metabolic signaling through PyK is essential for the integrity of motor neuron axons and synapses, and that its disruption activates both neurodegenerative and neuroprotective mechanisms
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spelling doaj-art-d3b8092ec14b443f8ec00ab24e2fd5182025-08-20T03:50:06ZengElsevierMolecular Metabolism2212-87782025-08-019810218710.1016/j.molmet.2025.102187Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protectionThomas J. Waller0Catherine A. Collins1Monica Dus2Molecular, Cellular, and Developmental Biology Department, College of Literature, Science, and the Arts, The University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding authors.Molecular, Cellular, and Developmental Biology Department, College of Literature, Science, and the Arts, The University of Michigan, Ann Arbor, MI, 48109, USA; Department of Neurosciences, Case Western Reserve University, Cleveland, OH, 44106, USAMolecular, Cellular, and Developmental Biology Department, College of Literature, Science, and the Arts, The University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding authors.Objective: Metabolic disruption is a central feature to many neurodegenerative diseases. Despite this, many gaps exist in our understanding of how these perturbations link to the mechanisms of neural disease. In this study, we sought to understand how genetically-controlled, cell-specific loss of pyruvate kinase (PyK) impacts motor neuron synaptic integrity and how the canonical neurodegenerative proteins DLK and SARM1 respond to this break in homeostasis. Methods: This study made use of the genetically-tractable Drosophila melanogaster to cell-specifically express proteins (via the GAL4/UAS binary system), knockdown gene transcripts (via RNA interference), and knockout gene loci (via guide RNA-directed Cas9). Synaptic and axonal degeneration were measured through immunohistochemistry, microscopy, and blinded scoring of fly larvae at both early and later 3rd instar stages to test for progressive phenotypes. Nervous system injury through a physical nerve crush assay was used to assay functional outcomes of protective stress responses. Results: We found that knockdown or knockout of PyK results in progressive axonal and synaptic degeneration, dependent on signaling through DLK and SARM1. This degeneration is preceded by nuclear transcriptional activation by DLK and the downstream AP-1 transcription factor Fos. We also found evidence of a neuroprotective response through injury of PyK-deficient axons (before progressive degeneration has occurred), which results in delayed Wallerian degeneration. This delay shows dependence on DLK and Fos, and coincides with reduced axonal localization of SARM1 whose overexpression fully restores degeneration speed. Conclusions: These data support a rheostat model of DLK signaling that both promotes and inhibits axon degeneration in response to metabolic disruption. This rheostat likely converges on regulation of SARM1, which is required for the progressive synapse loss following PyK, but also abolishes the protective delay in injury-induced Wallerian degeneration when overexpressed. Overall, we conclude that metabolic signaling through PyK is essential for the integrity of motor neuron axons and synapses, and that its disruption activates both neurodegenerative and neuroprotective mechanismshttp://www.sciencedirect.com/science/article/pii/S2212877825000948NeurodegenerationGlycolysisMetabolismAxonSynapseStress response
spellingShingle Thomas J. Waller
Catherine A. Collins
Monica Dus
Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
Molecular Metabolism
Neurodegeneration
Glycolysis
Metabolism
Axon
Synapse
Stress response
title Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
title_full Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
title_fullStr Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
title_full_unstemmed Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
title_short Pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
title_sort pyruvate kinase deficiency links metabolic perturbations to neurodegeneration and axonal protection
topic Neurodegeneration
Glycolysis
Metabolism
Axon
Synapse
Stress response
url http://www.sciencedirect.com/science/article/pii/S2212877825000948
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