Proteomic identification of differentially expressed genes in neural stem cells and neurons differentiated from embryonic stem cells of cynomolgus monkey (Macaca fascicularis) in vitro

Akama, Kuniko; Horikoshi, Tomoe; Nakayama, Takashi; Otsu, Masahiro; Imaizumi, Noriaki; Nakamura, Megumi; Toda, Tosifusa; Inuma, Michiko; Hirano, Hisashi; Kondo, Yasushi; Suzuki, Yutaka; Inoue, Nozomu

doi:10.1016/j.bbapap.2010.10.009

Cited by 15 publications

(7 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, other groups have also reported that PHB proteins are highly expressed in primate ES cells, including in humans [12], [13]. However, the functions of PHBs in ES cells are still unknown.…”

Section: Resultsmentioning

confidence: 99%

Prohibitin 2 Regulates the Proliferation and Lineage-Specific Differentiation of Mouse Embryonic Stem Cells in Mitochondria

et al. 2014

View full text Add to dashboard Cite

BackgroundThe pluripotent state of embryonic stem (ES) cells is controlled by a network of specific transcription factors. Recent studies also suggested the significant contribution of mitochondria on the regulation of pluripotent stem cells. However, the molecules involved in these regulations are still unknown.Methodology/Principal FindingsIn this study, we found that prohibitin 2 (PHB2), a pleiotrophic factor mainly localized in mitochondria, is a crucial regulatory factor for the homeostasis and differentiation of ES cells. PHB2 was highly expressed in undifferentiated mouse ES cells, and the expression was decreased during the differentiation of ES cells. Knockdown of PHB2 induced significant apoptosis in pluripotent ES cells, whereas enhanced expression of PHB2 contributed to the proliferation of ES cells. However, enhanced expression of PHB2 strongly inhibited ES cell differentiation into neuronal and endodermal cells. Interestingly, only PHB2 with intact mitochondrial targeting signal showed these specific effects on ES cells. Moreover, overexpression of PHB2 enhanced the processing of a dynamin-like GTPase (OPA1) that regulates mitochondrial fusion and cristae remodeling, which could induce partial dysfunction of mitochondria.Conclusions/SignificanceOur results suggest that PHB2 is a crucial mitochondrial regulator for homeostasis and lineage-specific differentiation of ES cells.

show abstract

Section: Resultsmentioning

confidence: 99%

Prohibitin 2 Regulates the Proliferation and Lineage-Specific Differentiation of Mouse Embryonic Stem Cells in Mitochondria

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Each group contained 5 samples factor, with overexpressing DPYSL2 promoting dendritic growth [4]. The hyperphosphorylation of DPYSL2 may induce neuronal apoptosis in the development of Alzheimer's disease [2,8]. Additionally, Akama revealed in 2011 that DPYSLY2 was upregulated in the process of NSCs differentiating into neurons by proteomic identification in the cynomolgus monkey [2].…”

Section: Discussionmentioning

confidence: 99%

“…The hyperphosphorylation of DPYSL2 may induce neuronal apoptosis in the development of Alzheimer's disease [2,8]. Additionally, Akama revealed in 2011 that DPYSLY2 was upregulated in the process of NSCs differentiating into neurons by proteomic identification in the cynomolgus monkey [2]. Later, it was emphasized that preconditioning with Ginkgo biloba (EGb 761(R)) promoted neuroprotection through HO1 and CRMP2 in rats with brain ischemia [34].…”

Section: Discussionmentioning

confidence: 99%

DPYSL2 is a novel regulator for neural stem cell differentiation in rats: revealed by Panax notoginseng saponin administration

et al. 2020

View full text Add to dashboard Cite

Background: The limited neuronal differentiation of the endogenous or grafted neural stem cells (NSCs) after brain injury hampers the clinic usage of NSCs. Panax notoginseng saponins (PNS) were extensively used for their clinical value, such as in controlling blood pressure, blood glucose, and inhibiting neuronal apoptosis and enhancing neuronal protection, but whether or not it exerts an effect in promoting neuronal differentiation of the endogenous NSCs is completely unclear and the potential underlying mechanism requires further exploration. Methods: Firstly, we determined whether PNS could successfully induce NSCs to differentiate to neurons under the serum condition. Mass spectrometry and quantitative polymerase chain reaction (Q-PCR) were then performed to screen the differentially expressed proteins (genes) between the PNS + serum and serum control group, upon which dihydropyrimidinase-like 2 (DPYSL2), a possible candidate, was then selected for the subsequent research. To further investigate the actual role of DPYSL2 in the NSC differentiation, DPYSL2-expressing lentivirus was employed to obtain DPYSL2 overexpression in NSCs. DPYSL2-knockout rats were constructed to study its effects on hippocampal neural stem cells. Immunofluorescent staining was performed to identify the differentiation direction of NSCs after 7 days from DPYSL2 transfection, as well as those from DPYSL2-knockout rats. Results: Seven differentially expressed protein spots were detected by PD Quest, and DPYSL2 was found as one of the key factors of NSC differentiation in a PNS-treated condition. The results of immunostaining further showed that mainly Tuj1 and GFAP-positive cells increased in the DPYSL2-overexpressed group, while both were depressed in the hippocampal NSCs in the DPYSL2-knockout rat. Conclusions: The present study revealed that the differentiation direction of NSCs could be enhanced through PNS administration, and the DPYSL2 is a key regulator in promoting NSC differentiation. These results not only emphasized the effect of PNS but also indicated DPYSL2 could be a novel target to enhance the NSC differentiation in future clinical trials.

show abstract

“…Dihydropyrimidinase-related protein 2, (DPYSL2), also known as collapsin response mediator protein 2 (CRMP-2), and isocitrate dehydrogenase [NADP] (IDH2), seem to be key nodes in the networks. DPYSL2 is a plasma membrane associated protein, involved in neuronal differentiation, as well as in axon growth and guidance, neuronal growth cone collapse, and cell migration (62) and has been also reported to be up-regulated in NSC in monkey (63) and rat (64). DPYSL4 and 5 are proteins from the same family as DYSL2 that are also observed in this network.…”

Section: Fig 2 Experimental Setupmentioning

confidence: 99%

Comprehensive Quantitative Comparison of the Membrane Proteome, Phosphoproteome, and Sialiome of Human Embryonic and Neural Stem Cells

Melo‐Braga

Schulz

Liu

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Human embryonic stem cells (hESCs) can differentiate into neural stem cells (NSCs), which can further be differentiated into neurons and glia cells. Therefore, these cells have huge potential as source for treatment of neurological diseases. Membrane-associated proteins are very important in cellular signaling and recognition, and their function and activity are frequently regulated by posttranslational modifications such as phosphorylation and glycosylation. To obtain information about membraneassociated proteins and their modified amino acids potentially involved in changes of hESCs and NSCs as well as to investigate potential new markers for these two cell stages, we performed large-scale quantitative membrane-proteomic of hESCs and NSCs. This approach employed membrane purification followed by peptide dimethyl labeling and peptide enrichment to study the membrane subproteome as well as changes in phosphorylation and sialylation between hESCs and NSCs. Combining proteomics and modification specific proteomics we identified a total of 5105 proteins whereof 57% contained transmembrane domains or signal peptides. The enrichment strategy yielded a total of 10,087 phosphorylated peptides in which 78% of phosphopeptides were identified with >99% confidence in site assignment and 1810 unique formerly sialylated N-linked glycopeptides. Several proteins were identified as significantly regulated in hESCs and NSC, including proteins involved in the early embryonic and neural development. In the latter group of proteins, we could identify potential NSC markers as Pluripotent embryonic stem cell (ESC)1 -derived neural stem cells (NSCs) can differentiate into neurons and glia cells of the central nervous system (1), including specialized neuron types like dopaminergic, representing a potential source for treatment of neurological diseases, such as ParkinsonЈs disease. Therefore, a better understanding of the cellular processes behind the changes of hESCs into NSCs, including solid markers for each cell type, is fundamental to move forward with a successful regenerative cell therapy and to investigate the early human neurogenesis processes.Many markers have been reported for the two types of stem cells (2, 3), however several of these markers are also identified in other stem or progenitor cells such as CD133 (Prominin-1) (4). Discovery of cell surface specific markers for differentiated stem cells is highly relevant for future clinical applications. In particular being able to distinguish the developmental stages of the differentiation from parental stem cells to fully mature cells would allow a correct manipulation and isolation of the cell type of interest. Moreover, such study 1 The abbreviations used are: CID, collision-induced dissociation; FDR, false discovery rate; FLR, false localization rate; HCD, highenergy collision induced dissociation; hESC, human embryonic stem cell; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MSA, multistage activation; NSC, neural stem cell; PTM, post-translational modificat...

show abstract

Proteomic identification of differentially expressed genes in neural stem cells and neurons differentiated from embryonic stem cells of cynomolgus monkey (Macaca fascicularis) in vitro

Cited by 15 publications

References 50 publications

Prohibitin 2 Regulates the Proliferation and Lineage-Specific Differentiation of Mouse Embryonic Stem Cells in Mitochondria

Prohibitin 2 Regulates the Proliferation and Lineage-Specific Differentiation of Mouse Embryonic Stem Cells in Mitochondria

DPYSL2 is a novel regulator for neural stem cell differentiation in rats: revealed by Panax notoginseng saponin administration

Comprehensive Quantitative Comparison of the Membrane Proteome, Phosphoproteome, and Sialiome of Human Embryonic and Neural Stem Cells

Contact Info

Product

Resources

About