Abstract
Autophagy is a catabolic pathway for the degradation and recycling of intracellular components, contributing to maintain cell homeostasis. Changes in autophagy activity can be monitored by a variety of biochemical and functional assays that should be used in combination. Recently, it has been described that signaling from the primary cilium modulates autophagy. This novel and reciprocal interaction will impact diverse aspects of the cell biology in healthy and pathophysiological conditions. Here, we describe methods to monitor autophagy activity in cilia mutants, as well as the use of autophagy mutants to monitor ciliogenesis.
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References
Orhon I, Dupont N et al (2015) Autophagy and regulation of cilia function and assembly. Cell Death Differ 22(3):389–397
Pampliega O, Orhon I et al (2013) Functional interaction between autophagy and ciliogenesis. Nature 502(7470):194–200
Tang Z, Lin MG et al (2013) Autophagy promotes primary ciliogenesis by removing OFD1 from centriolar satellites. Nature 502(7470):254–257
Esteban-Martinez L, Boya P (2015) Autophagic flux determination in vivo and ex vivo. Methods (San Diego, CA) 75:79–86
Klionsky DJ, Abdalla FC et al (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8(4):445–544
Mizushima N, Yoshimori T (2007) How to interpret LC3 immunoblotting. Autophagy 3(6):542–545
Pankiv S, Clausen TH et al (2007) p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282(33):24131–24145
Pazour GJ, Dickert BL et al (2000) Chlamydomonas IFT88 and its mouse homologue, polycystic kidney disease gene tg737, are required for assembly of cilia and flagella. J Cell Biol 151(3):709–718
Peterson GL (1977) A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 83(2):346–356. doi:10.1016/0003-2697(77)90043-4
Schnell U, Dijk F et al (2012) Immunolabeling artifacts and the need for live-cell imaging. Nat Methods 9(2):152–158
Omran H, Loges NT (2009) Chapter 7 – Immunofluorescence staining of ciliated respiratory epithelial cells. In: Stephen MK, Gregory JP (eds) Methods in cell biology, vol 91. Academic, New York, NY, pp 123–133
Hung SY, Huang WP et al (2015) LC3 overexpression reduces Abeta neurotoxicity through increasing alpha7nAchR expression and autophagic activity in neurons and mice. Neuropharmacology 93:243–251
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Satir, B.H., Pampliega, O. (2016). Methods to Study Interactions Between Ciliogenesis and Autophagy. In: Satir, P., Christensen, S. (eds) Cilia. Methods in Molecular Biology, vol 1454. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3789-9_4
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DOI: https://doi.org/10.1007/978-1-4939-3789-9_4
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3787-5
Online ISBN: 978-1-4939-3789-9
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