Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity

Abstract As neurodegenerative conditions are increasingly linked to mitochondrial dysfunction, methods for studying brain cell metabolism at high spatial resolution are needed to elucidate neurodegeneration mechanisms. Two-photon excited fluorescence (TPEF) imaging is a non-destructive, high-resolut...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Emily Stuntz, Yusi Gong, Disha Sood, Volha Liaudanskaya, Dimitra Pouli, Kyle P. Quinn, Carlo Alonzo, Zhiyi Liu, David L. Kaplan, Irene Georgakoudi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/892df3322ed645709c32c8c32e640f6a
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:892df3322ed645709c32c8c32e640f6a
record_format	dspace
spelling	oai:doaj.org-article:892df3322ed645709c32c8c32e640f6a2021-12-02T16:06:43ZEndogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity10.1038/s41598-017-01015-92045-2322https://doaj.org/article/892df3322ed645709c32c8c32e640f6a2017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01015-9https://doaj.org/toc/2045-2322Abstract As neurodegenerative conditions are increasingly linked to mitochondrial dysfunction, methods for studying brain cell metabolism at high spatial resolution are needed to elucidate neurodegeneration mechanisms. Two-photon excited fluorescence (TPEF) imaging is a non-destructive, high-resolution technique for studying cell metabolism via endogenous fluorescence of reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD). We employed TPEF to study the metabolism of primary rat astrocyte and neuronal cultures under normal growth conditions and in response to manganese (Mn) treatment. Histograms of pixel-wise optical redox ratio, defined as FAD/(FAD + NAD(P)H), revealed three distinct redox distributions and significant differences in their relative weights between astrocytes and neurons. When treated with Mn, both cell types exhibited redox ratio shifts consistent with increased oxidative stress. However, the manner in which the redox distributions was affected was distinct for the two cell types. Furthermore, NAD(P)H fluorescence lifetime imaging revealed an increase in bound NAD(P)H fraction upon Mn treatment for neurons, consistent with enhanced apoptosis. Astrocytes showed a decrease in bound fraction, possibly due to a shift towards glycolytic metabolism in response to impaired respiration. These results exhibit TPEF’s utility for characterizing detailed metabolic changes of different brain cell types in response to neurotoxins.Emily StuntzYusi GongDisha SoodVolha LiaudanskayaDimitra PouliKyle P. QuinnCarlo AlonzoZhiyi LiuDavid L. KaplanIrene GeorgakoudiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Emily Stuntz Yusi Gong Disha Sood Volha Liaudanskaya Dimitra Pouli Kyle P. Quinn Carlo Alonzo Zhiyi Liu David L. Kaplan Irene Georgakoudi Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
description	Abstract As neurodegenerative conditions are increasingly linked to mitochondrial dysfunction, methods for studying brain cell metabolism at high spatial resolution are needed to elucidate neurodegeneration mechanisms. Two-photon excited fluorescence (TPEF) imaging is a non-destructive, high-resolution technique for studying cell metabolism via endogenous fluorescence of reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD). We employed TPEF to study the metabolism of primary rat astrocyte and neuronal cultures under normal growth conditions and in response to manganese (Mn) treatment. Histograms of pixel-wise optical redox ratio, defined as FAD/(FAD + NAD(P)H), revealed three distinct redox distributions and significant differences in their relative weights between astrocytes and neurons. When treated with Mn, both cell types exhibited redox ratio shifts consistent with increased oxidative stress. However, the manner in which the redox distributions was affected was distinct for the two cell types. Furthermore, NAD(P)H fluorescence lifetime imaging revealed an increase in bound NAD(P)H fraction upon Mn treatment for neurons, consistent with enhanced apoptosis. Astrocytes showed a decrease in bound fraction, possibly due to a shift towards glycolytic metabolism in response to impaired respiration. These results exhibit TPEF’s utility for characterizing detailed metabolic changes of different brain cell types in response to neurotoxins.
format	article
author	Emily Stuntz Yusi Gong Disha Sood Volha Liaudanskaya Dimitra Pouli Kyle P. Quinn Carlo Alonzo Zhiyi Liu David L. Kaplan Irene Georgakoudi
author_facet	Emily Stuntz Yusi Gong Disha Sood Volha Liaudanskaya Dimitra Pouli Kyle P. Quinn Carlo Alonzo Zhiyi Liu David L. Kaplan Irene Georgakoudi
author_sort	Emily Stuntz
title	Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
title_short	Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
title_full	Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
title_fullStr	Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
title_full_unstemmed	Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity
title_sort	endogenous two-photon excited fluorescence imaging characterizes neuron and astrocyte metabolic responses to manganese toxicity
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/892df3322ed645709c32c8c32e640f6a
work_keys_str_mv	AT emilystuntz endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT yusigong endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT dishasood endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT volhaliaudanskaya endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT dimitrapouli endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT kylepquinn endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT carloalonzo endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT zhiyiliu endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT davidlkaplan endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity AT irenegeorgakoudi endogenoustwophotonexcitedfluorescenceimagingcharacterizesneuronandastrocytemetabolicresponsestomanganesetoxicity
_version_	1718384912751394816

Endogenous Two-Photon Excited Fluorescence Imaging Characterizes Neuron and Astrocyte Metabolic Responses to Manganese Toxicity

Ejemplares similares