title: Multiphoton imaging of actin filament formation and mitochondrial energetics of human ACBT gliomas.
abstract: We studied the three-dimensional (3D) distribution of actin filaments and mitochondria in relation to ACBT glioblastoma cells migration. We embedded the cells in the spheroid form within collagen hydrogels and imaged them by in situ multiphoton microscopy (MPM). The static 3D overlay of the distribution of actin filaments and mitochondria provided a greater understanding of cell-to-cell and cell-to-substrate interactions and morphology. While imaging mitochondria to obtain ratiometric redox index based on cellular fluorescence from reduced nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide we observed differential sensitivity of the migrating ACBT glioblastoma cells to femtosecond laser irradiation employed in MPM. We imaged actin-green fluorescent protein fluorescence in live ACBT glioma cells and for the first time observed dynamic modulation of the pools of actin during migration in 3D. The MPM imaging, which probes cells directly within the 3D cancer models, could potentially aid in working out a link between the functional performance of mitochondria, actin distribution and cancer invasiveness.
authors.affiliation: Cell Molecular and Developmental Biology Program, University of California, Riverside, Riverside, CA, USA.
authors.forename: Yu-Jer
authors.initials: YJ
authors.lastname: Hwang
authors.forename: Nomiki
authors.initials: N
authors.lastname: Kolettis
authors.forename: Miso
authors.initials: M
authors.lastname: Yang
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Gillard
authors.forename: Edgar
authors.initials: E
authors.lastname: Sanchez
authors.forename: Chung-Ho
authors.initials: CH
authors.lastname: Sun
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Tromberg
authors.forename: Tatiana B
authors.initials: TB
authors.lastname: Krasieva
authors.forename: Julia G
authors.initials: JG
authors.lastname: Lyubovitsky
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D004734:Energy Metabolism; D005909:Glioblastoma / Q000473:pathology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton; D008928:Mitochondria / Q000378:metabolism*
chemical_list: D049452:Green Fluorescent Proteins