MicroRNA-502-3p modulates the GABA A subunits, synaptic proteins and mitochondrial morphology in hippocampal neurons
Abstract:
MicroRNA-502-3p (MiR-502-3p), a synapse enriched miRNA is considerably implicated in Alzheimer's disease (AD). Our previous study found the high expression level of miR-502-3p in AD synapses relative to controls. Further, miR-502-3p was found to modulate the GABAergic synapse function via modulating the GABA A receptor subunit α-1 (GABRA1) protein. The current study is attempted to examine the impact of miR-502-3p on other GABA subunit proteins, synaptic proteins, mitochondrial morphology and other hippocampal neuron genes. Mouse hippocampal neuronal (HT22) cells were transfected with miR-502-3p overexpression (OE) vector, miR-502-3p sponge (suppression) vector and scramble control vector. MiR-502-3p vectors transfection was confirmed by fluorescence microscopy. MiR-502-3p expression and GABRA1 expression was confirmed by qRT-PCR and miRNAScope in-situ hybridization. GABA A subunit and synaptic proteins were studied by immunoblotting analysis and mitochondrial morphology was analyzed by transmission electron microscopy (TEM) analysis. Further, Affymetrix gene array analysis was conducted in miR-502-3p overexpressed and suppressed cells. Our results observed that elevated miR-502-3p, negatively modulates the GABRA1 level. The levels of GABA A subunit and synaptic proteins were reduced by ectopic expression of miR-502-3p and increase by miR-502-3p suppression. The mitochondrial morphology was found to be improved in-terms of their number and length in miR-502-3p suppressed cells. Further, Gene array analysis unveiled the deregulation of several genes by miR-502-3p, which are associated with oxidative stress, immune response and synaptic function. These results provide new insights and an update to understand the biological roles of miR-502-3p in regulation of neuron function and synaptic activity.
Insights
MicroRNA-502-3p (miR-502-3p) impacts Alzheimer's disease by altering GABAergic synapses and neuronal function. Suppressing miR-502-3p improves mitochondrial morphology and synaptic protein levels, offering therapeutic potential.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- MicroRNA-502-3p (miR-502-3p) is upregulated in Alzheimer's disease (AD) synapses.
- Previous studies linked miR-502-3p to GABAergic synapse modulation via GABRA1.
- The role of miR-502-3p in other synaptic components and neuronal functions remains to be fully elucidated.
Purpose of the Study:
- To investigate the impact of miR-502-3p on additional GABA A receptor subunits, synaptic proteins, and mitochondrial morphology in hippocampal neurons.
- To explore the broader effects of miR-502-3p on hippocampal neuron gene expression.
- To provide updated insights into the biological roles of miR-502-3p in neuronal function and synaptic activity.
Main Methods:
- Transfection of mouse hippocampal neuronal (HT22) cells with miR-502-3p overexpression and suppression vectors.
- Confirmation of transfection and miRNA/gene expression using fluorescence microscopy, qRT-PCR, and miRNAScope in situ hybridization.
- Analysis of protein levels (GABA A subunits, synaptic proteins) via immunoblotting and mitochondrial morphology via transmission electron microscopy (TEM).
- Gene expression profiling using Affymetrix gene array analysis.
Main Results:
- Elevated miR-502-3p negatively modulated GABRA1 levels.
- miR-502-3p overexpression reduced GABA A subunit and synaptic protein levels; suppression increased them.
- Suppression of miR-502-3p improved mitochondrial number and length.
- Gene array analysis revealed miR-502-3p deregulation of genes involved in oxidative stress, immune response, and synaptic function.
Conclusions:
- miR-502-3p plays a significant role in regulating GABAergic synaptic function and protein expression in neurons.
- Modulation of miR-502-3p levels affects mitochondrial health and synaptic protein integrity.
- miR-502-3p influences neuronal function through pathways including oxidative stress and immune response, relevant to Alzheimer's disease pathogenesis.
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