Visualizing Arc mRNA transfer using single molecule fluorescence assays

Abstract

The transfer of genetic material between cells has been implicated in both maintaining neuronal function as well as the proliferation of many diseases and disorders, especially neurodegenerative disorders. Regulation of mRNA transfer through transport and localization is conducted by RNA binding proteins (RBPs), making RBPs responsible for control of gene expression. The activity‐regulated cytoskeleton‐associated protein, Arc, is an important component of maintaining neuronal synaptic plasticity which ensures the proper flow of cognitive information. In Alzheimer’s disease, an increase of Arc protein expression and localization in synaptic activity areas has been observed in compensating for memory and cognition loss. Although Arc is thought to be released from the synaptic cleft through Arc‐formed capsids to transfer Arc mRNA into new target cells, the mechanism of action is unknown. This project uses single molecule fluorescence methods to interrogate the dynamics of Arc allowing for visualization of the potential movement of Arc capsids/Arc mRNA and the transfer of this genetic information in human cells. Preliminary studies of the Arc using single molecule fluorescence in situ hybridization (smFISH) and live cell imaging techniques has shown a static trajectory of Arc diffusion. Further studies of the transport and transfer of mRNA from Arc capsids will allow for an understanding how this machinery works that can be the basis for the development of new drug delivery methods for diseases with RNA targets.Support or Funding InformationNIH grant number: 1T32GM1320461