Methods to detect apoptotic-like cell death in filamentous fungi.
Fungi are capable of undergoing apoptotic-like cell death, and display many of the characteristic features of apoptosis observed in multicellular organisms. These features include nuclear condensation, DNA fragmentation, translocation of phosphatidylserine from the cytoplasmic to the extracellular side of the plasma membrane, and increased levels of reactive oxygen species (ROS). Several assays can be used to detect apoptotic cells, and here we describe adaptations of assays such as TUNEL, Annexin V, and Evan's Blue for the investigation of apoptotic-like cell death in fungal hyphae. We also present approaches for monitoring nuclear condensation and production of ROS.
Department of Plant Pathology and Center for Plant Science Innovation, University of Nebraska, Lincoln, NE, USA.
Camile P
CP
Semighini
Steven D
SD
Harris
D017304:Annexin A5 / Q000378:metabolism*; D017209:Apoptosis; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D004249:DNA Damage; D053938:DNA Fragmentation; D005434:Flow Cytometry; D005658:Fungi / Q000254:growth & development / Q000378:metabolism*; D025301:Hyphae / Q000254:growth & development / Q000378:metabolism*; D020287:In Situ Nick-End Labeling / Q000379:methods*; D010718:Phosphatidylserines / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*
D017304:Annexin A5; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species