scholarly journals Differential Expression of Tyrosine Hydroxylase Protein and Apoptosis-Related Genes in Differentiated and Undifferentiated SH-SY5Y Neuroblastoma Cells Treated with MPP+

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Kawinthra Khwanraj ◽  
Chareerut Phruksaniyom ◽  
Suriyat Madlah ◽  
Permphan Dharmasaroja
Keyword(s):  
Tyrosine Hydroxylase ◽  
Differential Expression ◽  
Gradual Increase ◽  
Cell Model ◽  
Neuroblastoma Cells ◽  
Human Neuroblastoma ◽  
Dopaminergic Cell ◽  
Differentiated Cells ◽  
Undifferentiated Cells ◽  
Cell Plating

The human neuroblastoma SH-SY5Y cell line has been used as a dopaminergic cell model for Parkinson’s disease research. Whether undifferentiated or differentiated SH-SY5Y cells are more suitable remains controversial. This study aims to evaluate the expression of apoptosis-related mRNAs activated by MPP+and evaluate the differential expression of tyrosine hydroxylase (TH) in undifferentiated and retinoic acid- (RA-) induced differentiated cells. The western blot results showed a gradual decrease in TH in undifferentiated cells and a gradual increase in TH in differentiated cells from days 4 to 10 after cell plating. Immunostaining revealed a gradual increase in TH along with neuritic outgrowth in differentiated cells on days 4 and 7 of RA treatment. For the study on cell susceptibility to MPP+and the expression of apoptosis-related genes, MTT assay showed a decrease in cell viability to approximately 50% requiring 500 and 1000 μM of MPP+for undifferentiated and RA-differentiated cells, respectively. Using real-time RT-PCR, treatment with 500 μM MPP+led to significant increases in the Bax/Bcl-2 ratio, p53, and caspase-3 in undifferentiated cells but was without significance in differentiated cells. In conclusion, differentiated cells may be more suitable, and the shorter duration of RA differentiation may make the SH-SY5Y cell model more accessible.

Functional changes in sodium conductances in the human neuroblastoma cell line SH-SY5Y during in vitro differentiation

1996 ◽  
Vol 76 (6) ◽  
pp. 3920-3927 ◽  
Author(s):  
M. Toselli ◽  
P. Tosetti ◽  
V. Taglietti
Keyword(s):  
Neuroblastoma Cell ◽  
Neuroblastoma Cell Line ◽  
Human Neuroblastoma Cell ◽  
Sodium Currents ◽  
In Vitro Differentiation ◽  
Human Neuroblastoma ◽  
Human Neuroblastoma Cell Line ◽  
Differentiated Cells ◽  
Undifferentiated Cells

1. The electrophysiological properties of voltage-dependent sodium currents were studied in the human neuroblastoma cell line SH-SY5Y before and after in vitro differentiation with retinoic acid, with the use of the whole cell variant of the patch-clamp technique. 2. Voltage steps from a holding level of -90 mV to depolarizing potentials elicited, in both undifferentiated and differentiated cells, fast inward sodium currents that were full inactivating and tetrodotoxin sensitive. 3. In undifferentiated cells the current peaked at -10 mV, the half-activation potential was -35 mV, and the half-inactivation potential was -81 mV. In differentiated cells the current peaked at + 10 mV, the half-activation potential was -28 mV, and the half-inactivation potential was -56 mV. Moreover, the peak current amplitude was about a factor of 2 larger and inactivation kinetics was about a factor of 2 slower than in undifferentiated cells. 4. This diversity in sodium channel properties was related to differences in cell excitability. Under current-clamp conditions, intracellular injection of rectangular depolarizing current stimuli from a hyperpolarized membrane potential of about -100 mV elicited graded and weak regenerative responses in undifferentiated cells, whereas overshooting action potentials with faster rising phases could be elicited in differentiated cells.

Differential expression of adrenomedullin and its receptor component, receptor activity modifying protein (RAMP) 2 during hypoxia in cultured human neuroblastoma cells

Peptides ◽  
2001 ◽  
Vol 22 (11) ◽  
pp. 1795-1801 ◽  
Author(s):  
Tomomi Kitamuro ◽  
Kazuhiro Takahashi ◽  
Kazuhito Totsune ◽  
Masaharu Nakayama ◽  
Osamu Murakami ◽  
...  
Keyword(s):  
Differential Expression ◽  
Neuroblastoma Cells ◽  
Receptor Activity ◽  
Human Neuroblastoma Cells ◽  
Human Neuroblastoma ◽  
Receptor Component

Expression of Tyrosine Hydroxylase Increases the Resistance of Human Neuroblastoma Cells to Oxidative Insults

2009 ◽  
Vol 113 (1) ◽  
pp. 150-157 ◽  
Author(s):  
Jeferson L. Franco ◽  
Thaís Posser ◽  
Sarah L. Gordon ◽  
Larisa Bobrovskaya ◽  
Jennifer J. Schneider ◽  
...  
Keyword(s):  
Tyrosine Hydroxylase ◽  
Neuroblastoma Cells ◽  
Human Neuroblastoma Cells ◽  
Human Neuroblastoma

scholarly journals A New Model of Sensorial Neuron-Like Cells for HTS of Novel Analgesics for Neuropathic Pain

2018 ◽  
Vol 24 (2) ◽  
pp. 158-168 ◽  
Author(s):  
Antón L. Martínez ◽  
José Brea ◽  
Xavier Monroy ◽  
Manuel Merlos ◽  
Javier Burgueño ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Intracellular Calcium ◽  
High Throughput Screening ◽  
Cell Model ◽  
Chemical Library ◽  
Differentiated Cells ◽  
Drg Neuron ◽  
Undifferentiated Cells ◽  
Vitro Model

In this study we developed a new translational phenotypic in vitro model for high-throughput screening (HTS) of novel analgesics for treating neuropathic pain, in order to address the poor translation of traditional recombinant models. The immortalized dorsal root ganglia (DRG) neuron-like F11 cell line was selected based on its phenotype after differentiation. The acquisition of neuronal characteristics was evaluated by measuring the expression of TrkA as a DRG neuron marker ( p < 0.01) as well as by measuring the global neurite length ( p < 0.001). The response of F11 cells to ATP and KCl was obtained by measuring intracellular calcium concentration, dynamic mass redistribution, and membrane potential. A KCl-induced increase of intracellular calcium levels was chosen as the readout because of the better signal quality, higher reproducibility, and greater compatibility with HTS assay requirements compared with other methods. The response to KCl differed significantly between differentiated and undifferentiated cells ( p < 0.05), with an EC50 value of 5 mM in differentiated cells. The model was validated by screening the Prestwick Chemical Library. Five hits already proposed for neuropathic-related pain were identified, with IC50 values between 1 and 7 µM. This cell model provides a new tool for screening novel analgesics for the relief of neuropathic pain.

scholarly journals 6-Hydroxydopamine Induce Neurodegeneration In Terminally Differentiated SH-SY5Y Neuroblastoma Cells Via Enrichment Of The Nucleosomal Degradation Pathway: A Global Proteomic Approach

2021 ◽  
Author(s):  
Kasthuri Bai Magalingam ◽  
Sushela Devi Somanath ◽  
Premdass Ramdas ◽  
Nagaraja Haleagrahara ◽  
Ammu Kutty Radhakrishnan
Keyword(s):  
Lupus Erythematosus ◽  
Ribosomal Proteins ◽  
Molecular Mechanisms ◽  
Cell Model ◽  
Neuroblastoma Cells ◽  
Neural Cells ◽  
Down Regulation ◽  
Human Neuroblastoma ◽  
Proteomic Approach ◽  
6 Hydroxydopamine

Abstract The SH-SY5Y human neuroblastoma cells have been used for decades as an in vitro cell model of dopaminergic neurons to explore the underlying science of cellular and molecular mechanisms of neurodegeneration in Parkinson’s disease (PD). However, data revealing the protein expression changes in 6-OHDA induced cytotoxicity in differentiated SH-SY5Y cells remain void. Therefore, we investigated the differentially regulated proteins expressed in terminally differentiated SH-SY5Y cells (differ-SH-SY5Y neural cells) exposed to 6-hydroxydopamine (6-OHDA) using the LC-MS/MS technology and interpreted the data using the online bioinformatics databases such as PANTHER, STRING, and KEGG. Our studies demonstrated the neuronal development in differ-SH-SY5Y neural cells was implicated by the overexpression of proteins responsible for neurite formations such as calnexin (CANX) and calreticulin (CALR) besides significant down-regulation of ribosomal proteins. The enrichment of the KEGG ribosome pathway was detected with significant down-regulation (p < 0.05) of all the 21 ribosomal proteins in differ-SH-SY5Y neural cells compared with undifferentiated cells. Whereas in the PD model, the pathological changes induced by 6-OHDA were indicated by the presence of unfolded and misfolded proteins, which triggered the response of 10 kDa heat shock proteins, namely HSPE1 and HSPA9. Moreover, the 6-OHDA induced neurodegeneration in differ-SH-SY5Y neural cells also upregulated the voltage-dependent anion-selective channel protein 1 (VDAC1) protein and enriched the KEGG systemic lupus erythematosus pathway that was regulated by all 17 histone proteins (p < 0.05) in differ-SH-SY5Y neural cells. These results suggest the central event in the neurodegenerative mechanism induced by 6-OHDA in the PD cell model was the nucleosomal degradation in the KEGG-SLE pathway indicated by the increased histone release in the apoptotic cells.

scholarly journals ELASTICITY OF DIFFERENTIATED AND UNDIFFERENTIATED HUMAN NEUROBLASTOMA CELLS CHARACTERIZED BY ATOMIC FORCE MICROSCOPY

2015 ◽  
Vol 15 (05) ◽  
pp. 1550069 ◽  
Author(s):  
SHIJIA ZHAO ◽  
ALEX STAMM ◽  
JEONG SOON LEE ◽  
ALEXEI GRUVERMAN ◽  
JUNG YUL LIM ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Elastic Modulus ◽  
Neuroblastoma Cells ◽  
Human Neuroblastoma ◽  
Force Microscopy ◽  
Atomic Force ◽  
Neuroscience Research ◽  
Culture Model ◽  
Undifferentiated Cells

Human neuroblastoma (SH-SY5Y) cells, with its ability to differentiate into neurons, have been widely used as the in vitro cell culture model for neuroscience research, especially in studying the pathogenesis of Parkinson's disease (PD) and developing therapeutic strategies. Cellular elasticity could potentially serve as a biomarker to quantitatively distinguish undifferentiated and differentiated SH-SY5Y cells. The goal of this work is to characterize the retinoic acid (RA) induced alternations of elastic properties of SH-SY5Y cells using atomic force microscopy (AFM). The elasticity was measured at multiple points of a single cell. Results have shown that the differentiation of SH-SY5Y cell led to a larger elastic modulus, which is three times more than that of undifferentiated cells. A higher indentation rate applied during AFM measurements led to a larger elastic modulus of the cell. This work provides new insights into the differentiation process identified by the elasticity marker, which could be extended to investigate the function, health and ageing of cells.

scholarly journals Production of Modified C-Reactive Protein in U937-Derived Macrophages

2005 ◽  
Vol 230 (10) ◽  
pp. 762-770 ◽  
Author(s):  
Irina Ciubotaru ◽  
Lawrence A. Potempa ◽  
Rosemary C. Wander
Keyword(s):  
Cell Model ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Differentiated Cells ◽  
Undifferentiated Cells ◽  
The Media

Plasma C-reactive protein (CRP) has been proposed to be a strong independent predictor for cardiovascular disease. This circulating form of CRP (native CRP or nCRP) is well described. Recently, the existence of a conformationally distinct isoform of CRP (modified CRP or mCRP) has been reported. The relevance of each CRP isoform to atherosclerotic disease is unknown. The purpose of this study was to examine the natural expression of CRP in undifferentiated, differentiated, and stimulated macrophages, cells known to contribute to atherogenesis in vivo, and to determine whether transcribed CRP was expressed as nCRP or mCRP. Macrophages were generated from U937 monocytes using phorbol 12-myristate 13-acetate. Differentiated macrophages were further stimulated with lipopolysaccharides (LPS). In undifferentiated, differentiated, and stimulated cells, CRP expression was assessed by reverse transcription–polymerase chain reaction, and CRP protein production was measured by fluorescence microscopy and flow cytometry (cellular CRP) or high-sensitivity enzyme-linked immunosorbent assay (secreted CRP). CRP transcript was minimally expressed in undifferentiated cells. Expression increased markedly in macrophages during differentiation and was not affected by LPS at 24 hrs. Cellular CRP protein increased in a time-dependent manner after LPS stimulation, and this induction was mediated via interleukin (IL)-6 and IL-1β. A small amount of secreted CRP was detected in the media of differentiated cells, but it was not significantly increased after LPS stimulation. Using specific monoclonal antibodies, our data indicate that cellular CRP is directly translated as the mCRP rather than the nCRP isomer. These results indicate that U937-derived macrophages are a good cell model to further study the production of mCRP under conditions relevant for the atherogenic process.

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