A new method to measure autophagy flux in the nervous system.
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.
Neurounion Biomedical Foundation; Santiago, Chile; Biomedical Neuroscience Institute; Faculty of Medicine; University of Chile; Santiago, Chile.
Soledad
S
Matus
Biomedical Neuroscience Institute; Faculty of Medicine; University of Chile; Santiago, Chile; Program of Cellular and Molecular Biology; Institute of Biomedical Sciences; University of Chile; Santiago, Chile; Center for Molecular Studies of the Cell; Faculty of Medicine; University of Chile; Santiago, Chile.
Vicente
V
Valenzuela
Biomedical Neuroscience Institute; Faculty of Medicine; University of Chile; Santiago, Chile; Program of Cellular and Molecular Biology; Institute of Biomedical Sciences; University of Chile; Santiago, Chile; Center for Molecular Studies of the Cell; Faculty of Medicine; University of Chile; Santiago, Chile; Department of Immunology and Infectious Diseases; Harvard School of Public Health; Boston, MA USA.
Claudio
C
Hetz
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009420:Nervous System / Q000166:cytology* / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*
D008869:Microtubule-Associated Proteins; D049452:Green Fluorescent Proteins
LC3; and nervous system; autophagy flux; neurons