Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: March 29, 2011
Publication Date: December 21, 2011
Citation: Nadal, M., Gold, S.E. 2011. Assessment of autophagosome formation by transmission electron microscopy. In: Bolton, M.D., Thomma, B., editors. Methods in Molecular Biology--Plant Fungal Pathogens: Methods and Protocols. New York, NY: Humana Press. p. 481-489.
Interpretive Summary: Autophagy is used by fungi to degrade or remove components from cells. In this work we provide a protocol for analysis of autophagy in fungal cells by electron microscopy. Autophagy is an important aspect of fungal development and this protocol is intended to assist other researchers in their analysis of the process within fungal cells.
Autophagy is a complex degradative process by which cytosolic material, including organelles, is randomly sequestered within double-membrane bound vesicles termed autophagosomes and targeted for degradation. Initially described as a nutrient stress adaptation response, the process of autophagy is now recognized as a central mechanism involved in many developmental processes. In this chapter, we provide guidelines to assess the initial steps of autophagy by monitoring autophagic body vacuolar accumulation. We employed a standard electron microscopy approach to observe the vacuoles of nutrient stressed fungal cells.