The Proteome of the Differentiating Mesencephalic Progenitor Cell Line CSM14.1<i>In Vitro</i>

Weiß, Bertram; Haas, Stefan; Lessner, Grit; Mikkat, Stefan; Kreutzer, Michael; Glocker, Michael O.; Wree, Andreas; Schmitt, Oliver

doi:10.1155/2014/351821

Cited by 6 publications

(4 citation statements)

References 70 publications

(78 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…More relevant to our results, another study analyzing the proteome of the differentiated DA neurons derived from the mesencephalon of embryonic rat reported that actin and vimentin were up-regulated after the differentiation (Weiss et al. 2014). These observations, together with our findings that several proteins involved in cytoskeletal network are up-regulated during differentiation, emphasize the importance of DA neurons’ capability to migrate through the midbrain area.…”

Section: Discussionsupporting

confidence: 68%

A proteomic analysis of differentiating dopamine neurons derived from human embryonic stem cells

Ryu

Park

Lee

2019

Animal Cells and Systems

View full text Add to dashboard Cite

Human embryonic stem cells (hESC) are being exploited for potential use in cell transplantation due to their capacity for self-renewal and pluripotency. Dopamine (DA) neurons derived from hESC represent a promising source of cell replacement therapy for Parkinson’s disease (PD). While gene expression on the transcriptome level has been extensively studied, limited information is available for the proteome-level changes associated with DA neuron differentiation. Here we analyzed the proteome of differentiating DA neurons to search for the potential biomarkers to assess the efficiency of differentiation. Although the proteome profile of DA neurons did not exhibit significant changes, a number of cytoskeletal proteins including nuclear lamin, tropomyosin 1, and myosin light chain 1 were specifically up-regulated during differentiation. Expression analysis of the respective genes was also consistent with the proteome results. In addition, these differentially expressed proteins form protein interaction network with several PD-related proteins suggesting that they may play roles in PD pathogenesis as well as the maturation of DA neurons.

show abstract

Section: Discussionsupporting

confidence: 68%

A proteomic analysis of differentiating dopamine neurons derived from human embryonic stem cells

Ryu

Park

Lee

2019

Animal Cells and Systems

View full text Add to dashboard Cite

show abstract

“…NFL, marker of axonal differentiation encoding for intermediate filament proteins and involved in important functions such as cell communication, cell‐extracellular matrix interactions and neuronal outgrowth (Jang et al, 2010) was inhibited by itraconazole. Analogously the expression of GFAP, an intermediate filament protein, commonly used as an astrocyte marker, but also expressed in many non‐glial cells populations, including differentiating mesencephalic neurons (Weiss et al, 2014; Vernon and Griffin, 2005) was inhibited. We can reasonably assume that itraconazole reduced neuron outgrowth and caused significant neuronal shape alterations by interacting somehow with the differentiation pathway.…”

Section: Discussionmentioning

confidence: 99%

Effect of itraconazole on mouse mesencephalic neurons

Giorgi

Fausto

Pardini³

et al. 2015

Intl J of Devlp Neuroscience

View full text Add to dashboard Cite

Neuronal cells have complex geometrical shapes, long processes such as axons and dendrites, and as a response to specific stimuli, they go through polarized neuronal migration that influences connectivity and information processing. Recently, it has been discovered that itraconazole, a widely used systemic antifungal drug, has an effect on cell morphology, acting as an inhibitor of the morphogen Sonic Hedgehog (Shh) and of the mammalian target of rapamycin mTOR pathways. In this paper we evaluated the effect of itraconazole on mouse mesencephalic dopaminergic neurons following their neurite outgrowth and functional activity by [(3)H] DA uptake. Furthermore the expression of several neural markers, the activation of the mTOR and of the morphogenic Shh pathways in the neuronal population was examined. Our results show for the first time a strong alteration of neurons morphology and an inhibitory effect of differentiation by itraconazole, probably due to cholesterol trafficking reduction, mTOR and Shh pathways inhibition. The inhibition of mTOR and Shh pathways by this drug has also been found in other cellular systems such as endothelial cells and lung cancer cells, suggesting a conserved mechanism of intercellular communication. As itraconazole is currently involved in multiple human clinical trials as a prospective anticancer agent, the effect on neuronal differentiation should be taken into account.

show abstract

“…At roughly 25% confluence, cells were differentiated for 7 days by shifting the cultures to 39°C and 5% CO 2 and supplementing the medium with 1 g/ml insulin, 20 M dopamine, and 100 M ascorbic acid (Sigma). The CSM14.1 cell line is a rat mesencephalic progenitor cell line that was immortalized with a temperature-sensitive SV40 T antigen (a gift from Dale Bredeson, Buck Institute for Age Research, Novato, CA) (35,75). CSM14.1 cells were grown under the permissive conditions of 31°C and 5% CO 2 in DMEM with 10% FBS, penicillin, streptomycin, and glutamine.…”

Section: Methodsmentioning

confidence: 99%

NF-κB Activation Promotes Alphavirus Replication in Mature Neurons

Yeh

Park

Schultz

et al. 2019

J Virol

View full text Add to dashboard Cite

Alphaviruses are enveloped, positive-sense RNA viruses that are important causes of viral encephalomyelitis. Sindbis virus (SINV) infects the neurons of rodents and is a model for studying factors that regulate infection of neuronal cells. The outcome of alphavirus infection of the central nervous system is dependent on neuronal maturation status. Differentiated mature neurons survive and control viral replication better than undifferentiated immature neurons. The cellular factors involved in age-dependent susceptibility include higher levels of antiapoptotic and innate immune factors in mature neurons. Because NF-κB pathway activation is required for the initiation of both apoptosis and the host antiviral response, we analyzed the role of NF-κB during SINV infection of differentiated and undifferentiated rat neuronal cells. SINV infection induced canonical NF-κB activation, as evidenced by the degradation of IκBα and the phosphorylation and nuclear translocation of p65. Inhibition or deletion of the upstream IκB kinase substantially reduced SINV replication in differentiated but not in undifferentiated neuronal cells or mouse embryo fibroblasts. NF-κB inhibition did not affect the establishment of infection, replication complex formation, the synthesis of nonstructural proteins, or viral RNA synthesis in differentiated neurons. However, the translation of structural proteins was impaired, phosphorylation of the α subunit of eukaryotic translation initiation factor 2 (eIF2α) was decreased, and host protein synthesis was maintained, suggesting that NF-κB activation was involved in the regulation of translation during infection of mature neurons. Inhibition or deletion of double-stranded RNA-activated protein kinase (PKR) also decreased eIF2α phosphorylation, the translation of viral structural proteins, and virus production. Therefore, canonical NF-κB activation synergizes with PKR to promote SINV replication in differentiated neurons by facilitating viral structural protein translation. IMPORTANCE Mosquito-borne alphaviruses are a significant and growing cause of viral encephalomyelitis worldwide. The outcome of alphaviral neuronal infections is host age dependent and greatly affected by neuronal maturation status, with differentiated, mature neurons being more resistant to infection than undifferentiated, immature neurons. The biological factors that change during neuronal maturation and that influence the outcome of viral infection are currently only partially defined. These studies investigated the role of NF-κB in determining the outcome of alphaviral infection in mature and immature neurons. Inhibition of canonical NF-κB activation decreased alphavirus replication in mature neurons by regulating protein synthesis and limiting the production of the viral structural proteins but had little effect on viral replication in immature neurons or fibroblasts. Therefore, NF-κB is a signaling pathway that influences the maturation-dependent outcome of alphaviral infection in neurons and that highlights the importance of cellular context in determining the effects of signal pathway activation.

show abstract

The Proteome of the Differentiating Mesencephalic Progenitor Cell Line CSM14.1In Vitro

Cited by 6 publications

References 70 publications

A proteomic analysis of differentiating dopamine neurons derived from human embryonic stem cells

A proteomic analysis of differentiating dopamine neurons derived from human embryonic stem cells

Effect of itraconazole on mouse mesencephalic neurons

NF-κB Activation Promotes Alphavirus Replication in Mature Neurons

Contact Info

Product

Resources

About