Abstract
Autophagy is an important cellular homeostatic process, it degrades most long-lived proteins and some organelles by lysosome to provide raw materials for the survival of the cells during nutrient or energy deprivation condition. Autophagy is active in Sertoli cells and involved in many cellular processes. However, the precise role of autophagy in Sertoli cells is still largely unknown. Thus, the assessment of autophagy in Sertoli cells should be helpful for investigating the functional roles of autophagy in Sertoli cells. This chapter describes some methods for assessing autophagy in Sertoli cells, including detection of LC3 maturation/aggregation, transmission electron microscopy, half-life assessments of long-lived proteins, immunofluorescence microscopy, and co-localization of autophagy-targeted proteins with autophagy components or lysosomal proteins.
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References
Levine B, Kroemer G (2008) Autophagy in the pathogenesis of disease. Cell 132(1):27–42. https://doi.org/10.1016/j.cell.2007.12.018
Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140(3):313–326. https://doi.org/10.1016/j.cell.2010.01.028
Wang H et al (2014) Atg7 is required for acrosome biogenesis during spermatogenesis in mice. Cell Res 24(7):852–869. https://doi.org/10.1038/cr.2014.70
Tang Z et al (2013) Autophagy promotes primary ciliogenesis by removing OFD1 from centriolar satellites. Nature 502(7470):254–257. https://doi.org/10.1038/nature12606
Mizushima N, Komatsu M (2011) Autophagy: renovation of cells and tissues. Cell 147(4):728–741. https://doi.org/10.1016/j.cell.2011.10.026
Mizushima N, Levine B (2010) Autophagy in mammalian development and differentiation. Nat Cell Biol 12(9):823–830. https://doi.org/10.1038/ncb0910-823
Li WW, Li J, Bao JK (2012) Microautophagy: lesser-known self-eating. Cell Mol Life Sci 69(7):1125–1136. https://doi.org/10.1007/s00018-011-0865-5
Cuervo AM, Wong E (2014) Chaperone-mediated autophagy: roles in disease and aging. Cell Res 24(1):92–104. https://doi.org/10.1038/cr.2013.153
Mizushima N (2007) Autophagy: process and function. Genes Dev 21(22):2861–2873. https://doi.org/10.1101/gad.1599207
Mizushima N, Yoshimori T, Ohsumi Y (2011) The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol 27:107–132. https://doi.org/10.1146/annurev-cellbio-092910-154005
Yang Z, Klionsky DJ (2009) An overview of the molecular mechanism of autophagy. Curr Top Microbiol Immunol 335:1–32. https://doi.org/10.1007/978-3-642-00302-8_1
Yu X, Long YC, Shen HM (2015) Differential regulatory functions of three classes of phosphatidylinositol and phosphoinositide 3-kinases in autophagy. Autophagy 11(10):1711–1728. https://doi.org/10.1080/15548627.2015.1043076
Nakatogawa H, Suzuki K, Kamada Y, Ohsumi Y (2009) Dynamics and diversity in autophagy mechanisms: lessons from yeast. Nat Rev Mol Cell Biol 10(7):458–467. https://doi.org/10.1038/nrm2708
Carlsson SR, Simonsen A (2015) Membrane dynamics in autophagosome biogenesis. J Cell Sci 128(2):193–205. https://doi.org/10.1242/jcs.141036
Itakura E, Mizushima N (2010) Characterization of autophagosome formation site by a hierarchical analysis of mammalian Atg proteins. Autophagy 6(6):764–776
Karanasios E, Stapleton E, Manifava M, Kaizuka T, Mizushima N, Walker SA, Ktistakis NT (2013) Dynamic association of the ULK1 complex with omegasomes during autophagy induction. J Cell Sci 126(Pt 22):5224–5238. https://doi.org/10.1242/jcs.132415
Klionsky DJ et al (2016) Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12(1):1–222. https://doi.org/10.1080/15548627.2015.1100356
Griswold MD (1998) The central role of Sertoli cells in spermatogenesis. Semin Cell Dev Biol 9(4):411–416. https://doi.org/10.1006/scdb.1998.0203
Hai Y, Hou J, Liu Y, Yang H, Li Z, He Z (2014) The roles and regulation of Sertoli cells in fate determinations of spermatogonial stem cells and spermatogenesis. Semin Cell Dev Biol 29:66–75. https://doi.org/10.1016/j.semcdb.2014.04.007
Ma Y, Yang HZ, Xu LM, Huang YR, Dai HL, Kang XN (2015) Testosterone regulates the autophagic clearance of androgen binding protein in rat Sertoli cells. Sci Rep 5:8894. https://doi.org/10.1038/srep08894
Yefimova MG et al (2013) A chimerical phagocytosis model reveals the recruitment by Sertoli cells of autophagy for the degradation of ingested illegitimate substrates. Autophagy 9(5):653–666. https://doi.org/10.4161/auto.23839
Chen Y, Zhou Y, Wang X, Qian W, Han X (2013) Microcystin-LR induces autophagy and apoptosis in rat Sertoli cells in vitro. Toxicon 76:84–93. https://doi.org/10.1016/j.toxicon.2013.09.005
Eid N, Ito Y, Otsuki Y (2012) Enhanced mitophagy in Sertoli cells of ethanol-treated rats: morphological evidence and clinical relevance. J Mol Histol 43(1):71–80. https://doi.org/10.1007/s10735-011-9372-0
Liu C et al (2016) Autophagy is required for ectoplasmic specialization assembly in sertoli cells. Autophagy 12(5):814–832. https://doi.org/10.1080/15548627.2016.1159377
Mizushima N (2004) Methods for monitoring autophagy. Int J Biochem Cell Biol 36(12):2491–2502. https://doi.org/10.1016/j.biocel.2004.02.005
Tanida I, Ueno T, Kominami E (2008) LC3 and autophagy. Methods Mol Biol 445:77–88. https://doi.org/10.1007/978-1-59745-157-4_4
Eskelinen EL (2008) Fine structure of the autophagosome. Methods Mol Biol 445:11–28. https://doi.org/10.1007/978-1-59745-157-4_2
Acknowledgments
This work was supported by National Key R&D program of China (Grant No. 2016YFA0500901), and the National Nature Science of China (Grant No. 91519317, 91649202 and 31471277).
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Liu, C., Khan, J., Li, W. (2018). Assessing Autophagy in Sertoli Cells. In: Alves, M., Oliveira, P. (eds) Sertoli Cells. Methods in Molecular Biology, vol 1748. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7698-0_10
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DOI: https://doi.org/10.1007/978-1-4939-7698-0_10
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