title: Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells.
abstract: Analysis of the cellular distributions of coenzymes including NADH may aid in understanding a cells metabolic status. We altered serum concentration (0, 2, and 10%) to induce living myoblast cells to undergo the early stages of differentiation. Through microscopy and phasor-FLIM, we spatially mapped and identified variations in the distribution of free and bound NADH. Undifferentiated cells displayed abundant free NADH within the nucleus along with specific regions of more bound NADH. Complete serum starvation dramatically increased the fraction of bound NADH in the nucleus, indicating heightened requirement for transcriptional processes. In comparison, cells exposed to 2% serum exhibited intermediate free nuclear NADH fraction. Overall our results suggest an order of events in which a cell metabolic status alters significantly during the early stages of serum induced differentiation.
authors.affiliation: School of Science and Health, University of Western Sydney, Hawkesbury, New South Wales, Australia.
authors.forename: Belinda K
authors.initials: BK
authors.lastname: Wright
authors.forename: Laura M
authors.initials: LM
authors.lastname: Andrews
authors.forename: Mark R
authors.initials: MR
authors.lastname: Jones
authors.forename: Chiara
authors.initials: C
authors.lastname: Stringari
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Digman
authors.forename: Enrico
authors.initials: E
authors.lastname: Gratton
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry*; D003067:Coenzymes / Q000032:analysis; D008853:Microscopy; D032446:Myoblasts / Q000737:chemistry*; D009243:NAD / Q000032:analysis*; D051381:Rats; D013234:Stem Cells / Q000737:chemistry*
chemical_list: D003067:Coenzymes; D009243:NAD