title: Microvesicle and tunneling nanotube mediated intercellular transfer of g-protein coupled receptors in cell cultures.
abstract: 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. 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 D(2)R-CFP or A(2A)R-YFP. These two types of cells were co-cultured, and FRET analysis demonstrated simultaneously positive cells to the D(2)R-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. 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. 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.
authors.affiliation: Department of Biomolecular Sciences, University of Urbino Carlo Bo, 61029 Urbino, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Guescini
authors.forename: G
authors.initials: G
authors.lastname: Leo
authors.forename: S
authors.initials: S
authors.lastname: Genedani
authors.forename: C
authors.initials: C
authors.lastname: Carone
authors.forename: F
authors.initials: F
authors.lastname: Pederzoli
authors.forename: F
authors.initials: F
authors.lastname: Ciruela
authors.forename: D
authors.initials: D
authors.lastname: Guidolin
authors.forename: V
authors.initials: V
authors.lastname: Stocchi
authors.forename: M
authors.initials: M
authors.lastname: Mantuano
authors.forename: D O
authors.initials: DO
authors.lastname: Borroto-Escuela
authors.forename: K
authors.initials: K
authors.lastname: Fuxe
authors.forename: L F
authors.initials: LF
authors.lastname: Agnati
mesh_terms: D000818:Animals; D001692:Biological Transport; D019556:COS Cells; D002450:Cell Communication; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018613:Microscopy, Confocal; D043705:Receptor, Adenosine A2A / Q000378:metabolism*; D017448:Receptors, Dopamine D2 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D022161:Transport Vesicles / Q000378:metabolism*
chemical_list: C485182:Cyan Fluorescent Protein; D043705:Receptor, Adenosine A2A; D017448:Receptors, Dopamine D2; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins