Assessing Autophagy in Sertoli Cells

Methods Mol Biol. 2018:1748:113-128. doi: 10.1007/978-1-4939-7698-0_10.

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.

Keywords: Autophagy; Immunoblotting; Immunofluorescence; LC3; Sertoli cell; Transmission electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy*
  • Cells, Cultured
  • Lysosomes / metabolism*
  • Male
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Phagosomes / metabolism*
  • Sequestosome-1 Protein / metabolism
  • Sertoli Cells / metabolism
  • Sertoli Cells / pathology*

Substances

  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Sequestosome-1 Protein
  • Sqstm1 protein, mouse