Methods to Detect Apoptotic-Like Cell Death in Filamentous Fungi

Detail:

Series: Methods in Molecular Biology | Volume: 638 | Pub. Date: Jun-01-2010 | Page Range: 269-279 | DOI: 10.1007/978-1-60761-611-5_20

Year prepared: 2010

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.

Laboratory Protocols

Select from:Additional methods & referenceAflatoxinsAssaying antifungal levelsAssaying mycotoxinsAztec Labs Diagnostic testing ProtocolsBiochemical CalculatorsBiofilmBiotypingCell Component ExtractionClinical BreakpointsCollection of Clinical MaterialCosmidsDetection in clinical samplesDNA extractionEnzyme synthesisFungal Molecular BiologyGeneral Molecular BiologyGeneticsGrowth and StorageHistologyMethods for DNA sequencingMicroscopyModel systems: in vitroModel systems: in vivoMore protocol collectionsPCR  -RT-qPCRPhagocytosisPlanar liquid chromatographyPost Antifungal Effects (PAFE)Preparation of amphotericin pasteProtein methodsProtocol overviewsQuality control in mycologySampling Aspergillus sporesSecondary metabolitesSpecies/Strain identification  -In situ hydridizationStandard Operating Procedures:National Aspergillosis CentreSusceptibility testingVeterinaryVideo Protocols at the FGSCWhole blood fractionationwhole-cell screening