A new method to measure autophagy flux in the nervous system

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

“…953 The use of different serotypes of adeno-associated virus could be used to transduce other cell types at the CNS. 953,954 This methodology allows a reproducible and sensitive mCherry-GFP-LC3 detection, and a strong LC3 flux when animals are treated with autophagy inducers including rapamycin and trehalose. 955 Therefore, these combined strategies can be applied to monitor autophagy activity in mice and also determine autophagy alterations in animal models of diseases affecting the nervous system.…”

“…955 Therefore, these combined strategies can be applied to monitor autophagy activity in mice and also determine autophagy alterations in animal models of diseases affecting the nervous system. 953,954 Alternatively, confocal laser scanning microscopy, which makes it possible to obtain numerous sections and substantial data about spatial localization features, can be a suitable system for studying autophagic structures (especially for whole mount embryo in vivo analysis). 956 In addition, this method can be used to obtain quantitative data through densitometric analysis of fluorescent signals.…”

Autophagy and ethanol neurotoxicity

“…However, an increase in LC3-II may be caused by either a stimulation of autophagosomal formation or inhibition of autophagosomal degradation, or may result from autophagyindependent mechanisms. 36 Therefore, both generation and degradation of autophagosomes need to be carefully evaluated when LC3-II is used to measure autophagy activity. Ethanol increases LC3 lipidation (Fig.…”

RGS 9‐2 rescues dopamine D2 receptor levels and signaling in DYT 1 dystonia mouse models

“…The observation that changes in basal LC3‐II, p62, and LAMP‐2 levels did not reach statistical significance rules out an overt dysregulation of autophagy and supports a more confined impairment involving targeting of DRD2 to degradation. In line with this view, the mCherry‐GFP‐LC3 reporter (Castillo et al , ) was increased both in the autolysosomes and in other compartments of ChIs (Matus et al , ).…”