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Updated: Jun 17, 2026
Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies
Published on: April 22, 2017
PKC delta and NADPH oxidase in retinoic acid-induced neuroblastoma cell differentiation
Mariapaola Nitti1, Anna Lisa Furfaro, Claudia Cevasco
1Department of Experimental Medicine, University of Genoa, Italy. paonit@medicina.unige.it
Abstract:
The role of reactive oxygen species (ROS) in the regulation of signal transduction processes has been well established in many cell types and recently the fine tuning of redox signalling in neurons received increasing attention. With regard to this, the involvement of NADPH oxidase (NOX) in neuronal pathophysiology has been proposed but deserves more investigation. In the present study, we used SH-SY5Y neuroblastoma cells to analyse the role of NADPH oxidase in retinoic acid (RA)-induced differentiation, pointing out the involvement of protein kinase C (PKC) delta in the activation of NOX. Retinoic acid induces neuronal differentiation as revealed by the increased expression of MAP2, the decreased cell doubling rate, and the gain in neuronal morphological features and these events are accompanied by the increased expression level of PKC delta and p67(phox), one of the components of NADPH oxidase. Using DPI to inhibit NOX activity we show that retinoic acid acts through this enzyme to induce morphological changes linked to the differentiation. Moreover, using rottlerin to inhibit PKC delta or transfection experiments to overexpress it, we show that retinoic acid acts through this enzyme to induce MAP2 expression and to increase p67(phox) membrane translocation leading to NADPH oxidase activation. These findings identify the activation of PKC delta and NADPH oxidase as crucial steps in RA-induced neuroblastoma cell differentiation.
Insights
Retinoic acid (RA) drives neuroblastoma cell differentiation by activating protein kinase C (PKC) delta and NADPH oxidase (NOX). This pathway is crucial for neuronal development and may offer therapeutic targets for neurological disorders.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Reactive oxygen species (ROS) play a key role in cellular signal transduction.
- Redox signaling in neurons is increasingly recognized for its importance in neuronal function and pathophysiology.
- The specific role of NADPH oxidase (NOX) in neuronal differentiation requires further elucidation.
Purpose of the Study:
- To investigate the role of NADPH oxidase (NOX) in retinoic acid (RA)-induced differentiation of SH-SY5Y neuroblastoma cells.
- To identify the signaling molecules involved in NOX activation during neuronal differentiation.
- To elucidate the involvement of protein kinase C (PKC) delta in this process.
Main Methods:
- SH-SY5Y neuroblastoma cells were treated with retinoic acid (RA).
- NADPH oxidase (NOX) activity was inhibited using diphenyleneiodonium (DPI).
- Protein kinase C (PKC) delta was modulated using rottlerin or transfection for overexpression studies.
- Expression levels of MAP2 and p67(phox) were analyzed.
Main Results:
- Retinoic acid (RA) induced neuronal differentiation, characterized by increased MAP2 expression, reduced proliferation, and altered cell morphology.
- RA treatment led to increased expression of PKC delta and p67(phox), a component of NOX.
- Inhibition of NOX with DPI blocked RA-induced morphological changes.
- PKC delta activation was essential for RA-induced MAP2 expression and p67(phox) translocation, leading to NOX activation.
Conclusions:
- Activation of PKC delta and NADPH oxidase (NOX) are critical events in retinoic acid (RA)-induced neuronal differentiation of neuroblastoma cells.
- This study identifies a novel signaling pathway involving PKC delta and NOX in neuronal development.
- These findings highlight potential therapeutic targets for neurological conditions involving impaired neuronal differentiation.
