Measurement of autophagy flux in the nervous system in vivo.
Accurate methods to measure autophagic activity in vivo in neurons are not available, and most of the studies are based on correlative and static measurements of autophagy markers, leading to conflicting interpretations. Autophagy is an essential homeostatic process involved in the degradation of diverse cellular components including organelles and protein aggregates. Autophagy impairment is emerging as a relevant factor driving neurodegeneration in many diseases. Moreover, strategies to modulate autophagy have been shown to provide protection against neurodegeneration. Here we describe a novel and simple strategy to express an autophagy flux reporter in the nervous system of adult animals by the intraventricular delivery of adeno-associated viruses (AAV) into newborn mice. Using this approach we efficiently expressed a monomeric tandem mCherry-GFP-LC3 construct in neurons of the peripheral and central nervous system, allowing the measurement of autophagy activity in pharmacological and disease settings.
1] Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago, Chile [2] Center for Molecular Studies of the Cell, Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, University of Chile, Santiago, Chile.
K
K
Castillo
V
V
Valenzuela
S
S
Matus
M
M
Nassif
M
M
Oñate
Y
Y
Fuentealba
G
G
Encina
T
T
Irrazabal
G
G
Parsons
F A
FA
Court
B L
BL
Schneider
D
D
Armentano
C
C
Hetz
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D002460:Cell Line; D000229:Dependovirus / Q000378:metabolism; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D051379:Mice; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009420:Nervous System / Q000378:metabolism* / Q000648:ultrastructure; D012584:Sciatic Nerve / Q000378:metabolism / Q000648:ultrastructure; D013116:Spinal Cord / Q000378:metabolism / Q000648:ultrastructure
C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins