2-D DIGE identification of differentially expressed heterogeneous nuclear ribonucleoproteins and transcription factors during neural differentiation of human embryonic stem cells

Miguel Barthéléry, Amritha Jaishankar, Ugur Salli, Willard M. Freeman, Kent Vrana

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Neural stem cells (NSC) are progenitors that can give rise to all neural lineages. They are found in specific niches of fetal and adult brains and grow in vitro as non-adherent colonies, the neurospheres. These cells express the intermediate filament nestin, commonly considered an NSC marker. NSC can be derived as neurospheres from human embryonic stem cells (hESC). The mechanisms of cellular programming that hESC undergo during differentiation remain obscure. To investigate the commitment process of hESC during directed neural differentiation, we compared the nuclear proteomes of hESC and hESC-derived neurospheres. We used 2-D DIGE to conduct a quantitative comparison of hESC and NSC nuclear proteins and detected 1521 protein spots matched across three gels. Statistical analysis (ANOVA n = 3 with false discovery correction) revealed that only 2.1% of the densitometric signal was significantly changed. The ranges of average ratios varied from 1.2-to 11-fold at a statistically significant p-value, <0.05. MS/MS identified 15 regulated proteins previously shown to be involved in chromatin remodeling, mRNA processing and gene expression regulation. Notably, three members of the heterogeneous nuclear ribonucleoprotein family (AUF-1, and FBP-1 and FBP-2) register a 54, 70 and 99% increased expression, highlighting them as potential markers for NSC in vitro derivation. By contrast, Cpsf-6 virtually disappears with differentiation with an 11-fold drop in NSC, high-lighting this protein as a novel marker for undifferentiated ESC.

Original languageEnglish (US)
Pages (from-to)505-514
Number of pages10
JournalProteomics - Clinical Applications
Volume3
Issue number4
DOIs
StatePublished - May 14 2009

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Heterogeneous-Nuclear Ribonucleoproteins
Neural Stem Cells
Stem cells
Transcription Factors
Nestin
Proteins
Chromatin Assembly and Disassembly
Intermediate Filaments
Gene Expression Regulation
Proteome
Nuclear Proteins
Lighting
Nuclear Family
Gene expression regulation
Human Embryonic Stem Cells
Analysis of Variance
Gels
Messenger RNA
Brain
Analysis of variance (ANOVA)

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

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abstract = "Neural stem cells (NSC) are progenitors that can give rise to all neural lineages. They are found in specific niches of fetal and adult brains and grow in vitro as non-adherent colonies, the neurospheres. These cells express the intermediate filament nestin, commonly considered an NSC marker. NSC can be derived as neurospheres from human embryonic stem cells (hESC). The mechanisms of cellular programming that hESC undergo during differentiation remain obscure. To investigate the commitment process of hESC during directed neural differentiation, we compared the nuclear proteomes of hESC and hESC-derived neurospheres. We used 2-D DIGE to conduct a quantitative comparison of hESC and NSC nuclear proteins and detected 1521 protein spots matched across three gels. Statistical analysis (ANOVA n = 3 with false discovery correction) revealed that only 2.1{\%} of the densitometric signal was significantly changed. The ranges of average ratios varied from 1.2-to 11-fold at a statistically significant p-value, <0.05. MS/MS identified 15 regulated proteins previously shown to be involved in chromatin remodeling, mRNA processing and gene expression regulation. Notably, three members of the heterogeneous nuclear ribonucleoprotein family (AUF-1, and FBP-1 and FBP-2) register a 54, 70 and 99{\%} increased expression, highlighting them as potential markers for NSC in vitro derivation. By contrast, Cpsf-6 virtually disappears with differentiation with an 11-fold drop in NSC, high-lighting this protein as a novel marker for undifferentiated ESC.",
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2-D DIGE identification of differentially expressed heterogeneous nuclear ribonucleoproteins and transcription factors during neural differentiation of human embryonic stem cells. / Barthéléry, Miguel; Jaishankar, Amritha; Salli, Ugur; Freeman, Willard M.; Vrana, Kent.

In: Proteomics - Clinical Applications, Vol. 3, No. 4, 14.05.2009, p. 505-514.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 2-D DIGE identification of differentially expressed heterogeneous nuclear ribonucleoproteins and transcription factors during neural differentiation of human embryonic stem cells

AU - Barthéléry, Miguel

AU - Jaishankar, Amritha

AU - Salli, Ugur

AU - Freeman, Willard M.

AU - Vrana, Kent

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