Quantitative proteomics reveals oxygen-dependent changes in neuronal mitochondria affecting function and sensitivity to rotenone

Lance Villeneuve, Leann M. Tiede, Brenda Morsey, Howard S Fox

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Mitochondria are implicated in a variety of degenerative disorders and aging. Mitochondria are responsive to the oxygen in their environment, yet tissue culture is performed at atmospheric (21%) oxygen and not at physiological (1-11%) oxygen levels found in tissues. We employed imaging of mitochondrial probes, mass spectrometry, Western blots, and ATP assays of the human neuroblastoma cell-line SH-SY5Y and imaging of mitochondrial probes in human primary neurons under standard nonphysiological oxygen conditions (atmospheric) and under physiological oxygen levels in the nervous system to assess the impact of oxygen on mitochondrial function. SH-SY5Y cells cultured in physiological 5% oxygen exhibited the lowest reactive oxygen species (ROS) production, indicating that culture at 5% oxygen is favored; these results were mimicked in primary human cells. Mass spectrometric analysis revealed extensive mitochondrial proteomic alterations in SH-SY5Y cells based on oxygen culture condition. Among these, the rotenone-sensitive subunit of complex I NDUFV3 was increased in cells cultured at 5% oxygen. Rotenone is a Parkinson's disease-linked toxin, and correspondingly SH-SY5Y cells cultured at 5% oxygen also exhibited over 10 times greater sensitivity to rotenone than those cultured in atmospheric, 21%, oxygen. Our results indicate that neuronal mitochondria are responsive to oxygen levels and produce differential responses under different oxygen levels.

Original languageEnglish (US)
Pages (from-to)4599-4606
Number of pages8
JournalJournal of proteome research
Volume12
Issue number10
DOIs
StatePublished - Oct 4 2013

Fingerprint

Rotenone
Mitochondria
Proteomics
Oxygen
Cultured Cells
Imaging techniques
Tissue culture
Neurology
Neuroblastoma
Nervous System
Neurons

Keywords

  • bioenergetics/electron transfer complex
  • hyperoxia
  • hypoxia
  • mitochondria
  • neurodegeneration
  • oxidative stress

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Quantitative proteomics reveals oxygen-dependent changes in neuronal mitochondria affecting function and sensitivity to rotenone. / Villeneuve, Lance; Tiede, Leann M.; Morsey, Brenda; Fox, Howard S.

In: Journal of proteome research, Vol. 12, No. 10, 04.10.2013, p. 4599-4606.

Research output: Contribution to journalArticle

@article{f4b279b541124a22a7249deb016a2a35,
title = "Quantitative proteomics reveals oxygen-dependent changes in neuronal mitochondria affecting function and sensitivity to rotenone",
abstract = "Mitochondria are implicated in a variety of degenerative disorders and aging. Mitochondria are responsive to the oxygen in their environment, yet tissue culture is performed at atmospheric (21{\%}) oxygen and not at physiological (1-11{\%}) oxygen levels found in tissues. We employed imaging of mitochondrial probes, mass spectrometry, Western blots, and ATP assays of the human neuroblastoma cell-line SH-SY5Y and imaging of mitochondrial probes in human primary neurons under standard nonphysiological oxygen conditions (atmospheric) and under physiological oxygen levels in the nervous system to assess the impact of oxygen on mitochondrial function. SH-SY5Y cells cultured in physiological 5{\%} oxygen exhibited the lowest reactive oxygen species (ROS) production, indicating that culture at 5{\%} oxygen is favored; these results were mimicked in primary human cells. Mass spectrometric analysis revealed extensive mitochondrial proteomic alterations in SH-SY5Y cells based on oxygen culture condition. Among these, the rotenone-sensitive subunit of complex I NDUFV3 was increased in cells cultured at 5{\%} oxygen. Rotenone is a Parkinson's disease-linked toxin, and correspondingly SH-SY5Y cells cultured at 5{\%} oxygen also exhibited over 10 times greater sensitivity to rotenone than those cultured in atmospheric, 21{\%}, oxygen. Our results indicate that neuronal mitochondria are responsive to oxygen levels and produce differential responses under different oxygen levels.",
keywords = "bioenergetics/electron transfer complex, hyperoxia, hypoxia, mitochondria, neurodegeneration, oxidative stress",
author = "Lance Villeneuve and Tiede, {Leann M.} and Brenda Morsey and Fox, {Howard S}",
year = "2013",
month = "10",
day = "4",
doi = "10.1021/pr400758d",
language = "English (US)",
volume = "12",
pages = "4599--4606",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "10",

}

TY - JOUR

T1 - Quantitative proteomics reveals oxygen-dependent changes in neuronal mitochondria affecting function and sensitivity to rotenone

AU - Villeneuve, Lance

AU - Tiede, Leann M.

AU - Morsey, Brenda

AU - Fox, Howard S

PY - 2013/10/4

Y1 - 2013/10/4

N2 - Mitochondria are implicated in a variety of degenerative disorders and aging. Mitochondria are responsive to the oxygen in their environment, yet tissue culture is performed at atmospheric (21%) oxygen and not at physiological (1-11%) oxygen levels found in tissues. We employed imaging of mitochondrial probes, mass spectrometry, Western blots, and ATP assays of the human neuroblastoma cell-line SH-SY5Y and imaging of mitochondrial probes in human primary neurons under standard nonphysiological oxygen conditions (atmospheric) and under physiological oxygen levels in the nervous system to assess the impact of oxygen on mitochondrial function. SH-SY5Y cells cultured in physiological 5% oxygen exhibited the lowest reactive oxygen species (ROS) production, indicating that culture at 5% oxygen is favored; these results were mimicked in primary human cells. Mass spectrometric analysis revealed extensive mitochondrial proteomic alterations in SH-SY5Y cells based on oxygen culture condition. Among these, the rotenone-sensitive subunit of complex I NDUFV3 was increased in cells cultured at 5% oxygen. Rotenone is a Parkinson's disease-linked toxin, and correspondingly SH-SY5Y cells cultured at 5% oxygen also exhibited over 10 times greater sensitivity to rotenone than those cultured in atmospheric, 21%, oxygen. Our results indicate that neuronal mitochondria are responsive to oxygen levels and produce differential responses under different oxygen levels.

AB - Mitochondria are implicated in a variety of degenerative disorders and aging. Mitochondria are responsive to the oxygen in their environment, yet tissue culture is performed at atmospheric (21%) oxygen and not at physiological (1-11%) oxygen levels found in tissues. We employed imaging of mitochondrial probes, mass spectrometry, Western blots, and ATP assays of the human neuroblastoma cell-line SH-SY5Y and imaging of mitochondrial probes in human primary neurons under standard nonphysiological oxygen conditions (atmospheric) and under physiological oxygen levels in the nervous system to assess the impact of oxygen on mitochondrial function. SH-SY5Y cells cultured in physiological 5% oxygen exhibited the lowest reactive oxygen species (ROS) production, indicating that culture at 5% oxygen is favored; these results were mimicked in primary human cells. Mass spectrometric analysis revealed extensive mitochondrial proteomic alterations in SH-SY5Y cells based on oxygen culture condition. Among these, the rotenone-sensitive subunit of complex I NDUFV3 was increased in cells cultured at 5% oxygen. Rotenone is a Parkinson's disease-linked toxin, and correspondingly SH-SY5Y cells cultured at 5% oxygen also exhibited over 10 times greater sensitivity to rotenone than those cultured in atmospheric, 21%, oxygen. Our results indicate that neuronal mitochondria are responsive to oxygen levels and produce differential responses under different oxygen levels.

KW - bioenergetics/electron transfer complex

KW - hyperoxia

KW - hypoxia

KW - mitochondria

KW - neurodegeneration

KW - oxidative stress

UR - http://www.scopus.com/inward/record.url?scp=84885214824&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885214824&partnerID=8YFLogxK

U2 - 10.1021/pr400758d

DO - 10.1021/pr400758d

M3 - Article

C2 - 23971408

AN - SCOPUS:84885214824

VL - 12

SP - 4599

EP - 4606

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 10

ER -