Astrocyte-to-neuron transportation of enhanced green fluorescent protein in cerebral cortex requires F-actin dependent tunneling nanotubes

Tunneling nanotube (TNT), a dynamic cell–cell contact, is dependent on actin polymerization. TNTs are efficient in transporting ions, proteins and organelles intercellularly, which are important mechanisms in physiological and pathological processes. Reported studies on the existence and function of TNTs among neural cells focus on cultured cell for the convenience in detecting TNTs’ ultrastructure. In this study, the adeno-associated virus (AAV-GFAP-EGFP-p2A-cre) was injected into the cerebral cortex of knock-in mice ROSA26 GNZ. GFAP promoter initiated the expression of enhanced green fluorescent protein (EGFP) in infected astrocytes. At 10 days post injection (10 DPI), EGFP transferred from astrocytes in layer I–III to neurons in layer V. The dissemination of EGFP was not through endocytosis or exosome. Applying microscopes, we found that the intercellular transportation of EGFP through contact connection was F-actin dependent. Therefore, we concluded that EGFP transported from astrocytes to neurons in cortex via F-actin dependent TNTs. This study first proved that proteins transported intercellularly via TNTs in brain.