Microvesicle and tunneling nanotube mediated intercellular transfer of g-protein coupled receptors in cell cultures

作者：Guescini M; Leo G; Genedani S; Carone C; Pederzoli F; Ciruela F; Guidolin D; Stocchi V; Mantuano M; Borroto Escuela D O; Fuxe K; Agnati L F^*

来源：Experimental Cell Research, 2012, 318(5): 603-613.

DOI：10.1016/j.yexcr.2012.01.005

• Microvesicles
• Inter-cellular communication
• GPCRs
• Exosomes
• Synaptic modulation

Recent evidence shows that cells exchange collections of signals via microvesicles (MVs) and tunneling nano-tubes (TNTs). In this paper we have investigated whether in cell cultures GPCRs can be transferred by means of MVs and TNTs from a source cell to target cells. %26lt;br%26gt;Western blot, transmission electron microscopy and gene expression analyses demonstrate that A(2A) and D-2 receptors are present in released MVs. In order to further demonstrate the involvement of MVs in cell-to-cell communication we created two populations of cells (HEK293T and COS-7) transiently transfected with D2R-CFP or A(2A)R-YFP. These two types of cells were co-cultured, and FRET analysis demonstrated simultaneously positive cells to the D2R-CFP and A(2A)R-YFP. Fluorescence microscopy analysis also showed that GPCRs can move from one cell to another also by means of TNTs. %26lt;br%26gt;Finally, recipient cells pre-incubated for 24 h with A(2A)R positive MVs were treated with the adenosine A(2A) receptor agonist CGS-21680. The significant increase in cAMP accumulation clearly demonstrated that A(2A)Rs were functionally competent in target cells. %26lt;br%26gt;These findings demonstrate that A(2A) receptors capable of recognizing and decoding extracellular signals can be safely transferred via MVs from source to target cells.

• 出版日期2012-3-10