title: Monitoring Phosphatidic Acid Signaling in Breast Cancer Cells Using Genetically Encoded Biosensors.
abstract: Phospholipids are important signaling molecules that regulate cell proliferation, death, migration, and metabolism. Many phospholipid signaling cascades are altered in breast cancer. To understand the functions of phospholipid signaling molecules, genetically encoded phospholipid biosensors have been developed to monitor their spatiotemporal dynamics. Compared to other phospholipids, much less is known about the subcellular production and cellular functions of phosphatidic acid (PA), partially due to the lack of a specific and sensitive PA biosensor in the past. This chapter describes the use of a newly developed PA biosensor, PASS, in two applications: regular fluorescent microscopy and fluorescence lifetime imaging microscopy-Förster/fluorescence resonance energy transfer (FLIM-FRET). These protocols can be also used with other phospholipid biosensors.
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, 6431 Fannin St, Houston, TX, 77030, USA.
authors.forename: Maryia
authors.initials: M
authors.lastname: Lu
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, 6431 Fannin St, Houston, TX, 77030, USA.
authors.forename: Li Wei Rachel
authors.initials: LW
authors.lastname: Tay
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, 6431 Fannin St, Houston, TX, 77030, USA.
authors.forename: Jingquan
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, 6431 Fannin St, Houston, TX, 77030, USA. guangwei.du@uth.tmc.edu.
authors.forename: Guangwei
authors.initials: G
authors.lastname: Du
mesh_terms: D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000494:pharmacology; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005818:Genetic Engineering; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D010712:Phosphatidic Acids / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection
chemical_list: D010712:Phosphatidic Acids; D004815:Epidermal Growth Factor
keywords: Cancer; FLIM; FRET; Fluorescence lifetime imaging microscopy; Fluorescence resonance energy transfer; Immunofluorescence; PASS; Phosphatidic acid; Phospholipid
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