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Distinct Dynamics and Composition of Endogenous Neuronal RNA Granules

ABSTRACT: RNA granules are essential regulators of post-transcriptional gene expression, enabling mRNA transport, localization and local translation in neurons. The localized transcriptome is diverse; however, how different mRNAs are organized into granules for efficient localization and translation remains unknown. Here, we combine real time single RNA imaging with protein and RNA proximity labeling to investigate two endogenous neuronal mRNA granule populations, Actb and Arc, in stimulated primary hippocampal neurons. Using orthogonal RNA labeling systems in a dual knock-in mouse model, we show that Actb and Arc mRNAs are packaged into spatially segregated granules with distinct trafficking dynamics, localization kinetics, and responses to synaptic stimulation. While Actb granules displayed rapid and sustained localization, whereas Arc granules showed delayed, transient recruitment, consistent with their respective roles in structural and activity-dependent plasticity. Proximity labeling reveals that these granules are entirely distinct in their RNA signatures, despite sharing core RNA-binding proteins. Using motif analysis, we propose that shared cis-regulatory elements may drive selective co-packaging of mRNAs into unique granules. Together, these findings demonstrate that neuronal mRNAs are differentially sorted into molecularly and functionally distinct granules, providing a framework for understanding how precise spatio-temporal control of mRNA localization and translation is achieved across complex neuronal arbors.

ORGANISM(S): Mus musculus

PROVIDER: GSE326430 | GEO | 2026/04/01

REPOSITORIES: GEO

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