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Updated: Jul 11, 2026
Differentiation of a Human Neural Stem Cell Line on Three Dimensional Cultures, Analysis of MicroRNA and Putative Target Genes
Published on: April 12, 2015
MicroRNAs regulate synthesis of the neurotransmitter substance P in human mesenchymal stem cell-derived neuronal
Steven J Greco1, Pranela Rameshwar
1Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA.
Abstract:
MicroRNAs (miRNAs) are a class of 19- to 23-nt, small, noncoding RNAs, which bind the 3' UTR of target mRNAs to mediate translational repression in animals. miRNAs have been shown to regulate developmental processes, such as self-renewal of stem cells, neuronal differentiation, myogenesis, and cancer. A functional role of miRNAs in the regulation of neurotransmitter synthesis has yet to be ascribed. We used mesenchymal stem cells (MSCs) as a model to study miRNA-mediated neurotransmitter regulation in developing neuronal cells. MSCs are mesoderm-derived cells, primarily resident in adult bone marrow, which can generate functional neuronal cells. We have previously shown that human MSC-derived neuronal cells express the neurotransmitter gene, Tac1, but do not synthesize the gene's encoded peptide, the neurotransmitter substance P (SP), unless stimulated with the inflammatory mediator IL-1alpha. These findings suggested a potential role for miRNAs in the regulation of SP synthesis. Here, we report on the miRNA profile of undifferentiated human MSCs and MSC-derived neuronal cells by using miRNA-specific bioarrays. miRNAs that were increased in the neuronal cells and decreased after IL-1alpha stimulation were analyzed by the miRanda algorithm to predict Tac1 mRNA targets. Putative miR-130a, miR-206, and miR-302a binding sites were predicted within the 3' UTR of Tac1. Target validation using a luciferase reporter system confirmed the miR-130a and miR-206 sites. Specific inhibition of miR-130a and miR-206 in the neuronal cells resulted in SP synthesis and release. The studies provide a different approach in ascribing a new regulatory role for miRNAs in regulating neurotransmitter synthesis.
Insights
MicroRNAs (miRNAs) regulate neurotransmitter synthesis. Inhibiting miR-130a and miR-206 in stem cell-derived neurons stimulated substance P release, revealing a novel miRNA regulatory role.
Area of Science:
- Molecular Biology
- Neuroscience
- Stem Cell Biology
Background:
- MicroRNAs (miRNAs) are small noncoding RNAs regulating gene expression post-transcriptionally.
- While miRNAs regulate development and disease, their role in neurotransmitter synthesis is largely unknown.
- Mesenchymal stem cells (MSCs) can differentiate into functional neuronal cells and express neurotransmitter genes like Tac1.
Purpose of the Study:
- To investigate the role of miRNAs in regulating neurotransmitter synthesis, specifically substance P (SP), in MSC-derived neuronal cells.
- To identify specific miRNAs involved in the regulation of Tac1 gene expression and SP production.
Main Methods:
- Profiling of miRNA expression in undifferentiated MSCs and MSC-derived neuronal cells using miRNA-specific bioarrays.
- Bioinformatic analysis (miRanda algorithm) to predict miRNA binding sites on Tac1 mRNA 3' UTR.
- Luciferase reporter assays for target validation and functional studies involving miRNA inhibition.
Main Results:
- Specific miRNAs, including miR-130a and miR-206, were identified as potential regulators of Tac1.
- Luciferase assays confirmed miR-130a and miR-206 binding sites within the Tac1 3' UTR.
- Inhibition of miR-130a and miR-206 in neuronal cells led to increased synthesis and release of substance P.
Conclusions:
- This study demonstrates a novel regulatory role for specific miRNAs (miR-130a, miR-206) in controlling neurotransmitter synthesis.
- miRNAs can modulate the expression of genes involved in neurotransmitter production, offering new therapeutic targets.
- MSC-derived neuronal cells serve as a valuable model for studying miRNA-mediated regulation of neuronal function.
