A new method to measure autophagy flux in the nervous system
Open accessAbstract
A current need in the neuroscience field is a simple method to monitor autophagic activity in vivo in neurons. Until very recently, most reports have been based on correlative and static determinations of the expression levels of autophagy markers in the brain, generating conflicting interpretations. Autophagy is a fundamental process mediating the degradation of diverse cellular components, including organelles and protein aggregates at basal levels, whereas alterations in the process (i.e., autophagy impairment) operate as a pathological mechanism driving neurodegeneration in most prevalent diseases. We have recently described a new simple method to deliver and express an autophagy flux reporter through the peripheral and central nervous system of mice by the intracerebroventricular delivery of adeno-associated viruses (AAV) into newborn mice. We obtained a wide expression of a monomeric tandem mCherry-GFP-LC3 construct in neurons through the nervous system and demonstrated efficient and accurate measurements of LC3 flux after pharmacological stimulation of the pathway or in disease settings of axonal damage. Here we discuss the possible applications of this new method to assess autophagy activity in neurons in vivo.
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The GFP fluorescence produced by this AAV is sensitive to the acidic and/or proteolytic conditions of the lysosome lumen, while mCherry retains its fluorescence even at acidic pH.
“The GFP fluorescence produced by this AAV is sensitive to the acidic and/or proteolytic conditions of the lysosome lumen, while mCherry retains its fluorescence even at acidic pH (Matus et al., 2014).”
From Nilotinib restores memory function by preventing dopaminergic neuron degeneration in a mouse model of Alzheimer’s Disease (2021) · cites this paper as Matus et al., 2014
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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).
Klionsky, D. J., Abdelmohsen, K., Abe, A., Abedin, M. J., Abeliovich, H., Acevedo Arozena, A., Adachi, H., Adams, C. M., Adams, P. D., Adeli, K., et al. (2016) · Autophagy · doi:10.1080/15548627.2015.1100356 · Cited by 5,894 · see citing works
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“its fluorescence even at acidic pH (Matus et al., 2014).”
- Authors
- Matus, S., Valenzuela, V., & Hetz, C.
- Affiliations
- Neurounion Biomedical Foundation (CL)
- University of Chile (CL)
- Harvard University (US)
- Corresponding author
- Matus, Soledad
- Hetz, Claudio
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2014-03-17
- Volume
- 10
- Issue
- 4
- Pages
- 710-714
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC4091163
- OpenAlex
- W1986616897
- Cited by
- 29
- References
- 39
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 4 · 2020: 2 · 2019: 2 · 2018: 5 · 2017: 3 · 2016: 4 · 2015: 1 · 2014: 3
- FWCI
- 2.77
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Autophagy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (publishedVersion, other-oa)
- Americanae (AECID Library) (submittedVersion, other-oa)
- LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, other-oa)
- Universidad de Chile (submittedVersion, cc-by-nc-nd)
- Funders
- Muscular Dystrophy Association
- ALS Therapy Alliance
- Comisión Nacional de Investigación Científica y Tecnológica (21120411, C13S02, ACT1109, P09-015F, USA2013-0003)
- Fondo de Fomento al Desarrollo Científico y Tecnológico (D11I1007, 11121524, ACT1109)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Keywords
- autophagic flux, LC3, adeno-associated virus, intracerebroventricular administration, neurons, neurodegeneration, axonal injury
- MeSH terms
- Animals, Autophagy, Microtubule-Associated Proteins, Nervous System, Neurons, Green Fluorescent Proteins, Mice
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