Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells

Belinda K. Wright; Laura M. Andrews; Mark R. Jones; Chiara Stringari; Michelle A. Digman; Enrico Gratton

doi:10.1002/jemt.22121

Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells

Belinda K. Wright, Laura M. Andrews, Mark R. Jones, Chiara Stringari, Michelle A. Digman, Enrico Gratton

School of Science

Research output: Contribution to journal › Article › peer-review

35 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1717-1722
Number of pages	6
Journal	Microscopy Research and Technique
Volume	75
Issue number	12
DOIs	https://doi.org/10.1002/jemt.22121
Publication status	Published - 2012

Keywords

NAD (coenzyme)
fluorescence microscopy
myoblasts

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10.1002/jemt.22121

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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.",

keywords = "NAD (coenzyme), fluorescence microscopy, myoblasts",

author = "Wright, {Belinda K.} and Andrews, {Laura M.} and Jones, {Mark R.} and Chiara Stringari and Digman, {Michelle A.} and Enrico Gratton",

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T1 - Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells

AU - Wright, Belinda K.

AU - Andrews, Laura M.

AU - Jones, Mark R.

AU - Stringari, Chiara

AU - Digman, Michelle A.

AU - Gratton, Enrico

PY - 2012

Y1 - 2012

N2 - 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.

AB - 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.

KW - NAD (coenzyme)

KW - fluorescence microscopy

KW - myoblasts

UR - http://handle.uws.edu.au:8081/1959.7/518538

U2 - 10.1002/jemt.22121

DO - 10.1002/jemt.22121

M3 - Article

C2 - 23019116

SN - 1059-910X

VL - 75

SP - 1717

EP - 1722

JO - Microscopy Research and Technique

JF - Microscopy Research and Technique

IS - 12

ER -

Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells

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Keywords

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