Proteomic analysis of expression and protein interactions in a 6-hydroxydopamine-induced rat brain lesion model

Bokyung Park, Junyoung Yang, Nuri Yun, Kwang Min Choe, Byung K. Jin, Young J. Oh

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Parkinson's disease (PD) is the second most common neurodegenerative disorder caused by selective degeneration of the dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc). Although mitochondrial abnormality, oxidative stress and proteasomal dysfunction are recognized as major contributors to the progression of PD, there is a limited understanding of the key molecular events that provoke degeneration of DA neurons. Using a proteomic approach, we attempted to identify profiles of proteins with altered expression levels in rats following unilateral stereotaxic injection of 6-hydroxydopamine into the SNc. Protein expression profiles of these proteins in the substantia nigra and the striatum were made using two-dimensional gel electrophoresis in conjunction with a mass spectrometry. More than 70 identified proteins displayed significant differences in their temporal and spatial expression pattern between experimental and vehicle-operated control groups. Based on the identity of the proteins, we further searched for potential binding partners using biological databases available on the web and constructed a protein interaction network. Among several interconnected proteins in the network, we verified the interaction between prohibitin and the NADH-ubiquinone oxidoreductase 30. kDa subunit (NDUFS3 subunit; a mitochondrial complex I subunit) by co-immunoprecipitation. We also confirmed, using immunohistochemical localization, that both prohibitin and the NDUFS3 subunit were increased in the dying DA neurons, suggesting its potential role in regulating mitochondrial function in dying DA neurons. Furthermore, knockdown of prohibitin accelerated 6-hydroxydopamine-induced cell death in SH-SY5Y cells. Our results raise the possibility that interconnected proteins in the network may positively or negatively impact the progression of DA neuronal death.

Original languageEnglish
Pages (from-to)16-32
Number of pages17
JournalNeurochemistry International
Volume57
Issue number1
DOIs
Publication statusPublished - 2010 Aug 1

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Oxidopamine
Proteomics
Dopaminergic Neurons
Brain
Proteins
Parkinson Disease
Electron Transport Complex I
Protein Interaction Maps
Electrophoresis, Gel, Two-Dimensional
Substantia Nigra
Immunoprecipitation
Neurodegenerative Diseases
Mass Spectrometry
Oxidative Stress
Cell Death
Databases
Control Groups
Injections
prohibitin

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience
  • Cell Biology

Cite this

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title = "Proteomic analysis of expression and protein interactions in a 6-hydroxydopamine-induced rat brain lesion model",
abstract = "Parkinson's disease (PD) is the second most common neurodegenerative disorder caused by selective degeneration of the dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc). Although mitochondrial abnormality, oxidative stress and proteasomal dysfunction are recognized as major contributors to the progression of PD, there is a limited understanding of the key molecular events that provoke degeneration of DA neurons. Using a proteomic approach, we attempted to identify profiles of proteins with altered expression levels in rats following unilateral stereotaxic injection of 6-hydroxydopamine into the SNc. Protein expression profiles of these proteins in the substantia nigra and the striatum were made using two-dimensional gel electrophoresis in conjunction with a mass spectrometry. More than 70 identified proteins displayed significant differences in their temporal and spatial expression pattern between experimental and vehicle-operated control groups. Based on the identity of the proteins, we further searched for potential binding partners using biological databases available on the web and constructed a protein interaction network. Among several interconnected proteins in the network, we verified the interaction between prohibitin and the NADH-ubiquinone oxidoreductase 30. kDa subunit (NDUFS3 subunit; a mitochondrial complex I subunit) by co-immunoprecipitation. We also confirmed, using immunohistochemical localization, that both prohibitin and the NDUFS3 subunit were increased in the dying DA neurons, suggesting its potential role in regulating mitochondrial function in dying DA neurons. Furthermore, knockdown of prohibitin accelerated 6-hydroxydopamine-induced cell death in SH-SY5Y cells. Our results raise the possibility that interconnected proteins in the network may positively or negatively impact the progression of DA neuronal death.",
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Proteomic analysis of expression and protein interactions in a 6-hydroxydopamine-induced rat brain lesion model. / Park, Bokyung; Yang, Junyoung; Yun, Nuri; Choe, Kwang Min; Jin, Byung K.; Oh, Young J.

In: Neurochemistry International, Vol. 57, No. 1, 01.08.2010, p. 16-32.

Research output: Contribution to journalArticle

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