Cilia and the exocyst are involved in urinary extracellular vesicle production and content

ABSTRACT: A recently proposed model of cell-cell communication, called “urocrine signaling”, hypothesizes that small (50-150 mm) extracellular vesicles (EVs) carry signals to “downstream” kidney tubule cells. We previously showed that the exocyst, a highly-conserved eight-protein (Exoc1-8) trafficking complex, is necessary for ciliogenesis, we show that the EV number released from MDCK cells was increased in EXOC5 OE, while it was decreased in Exoc5 KD, compared to control. To determine how Exoc5 changes the composition of EVs, we isolated EVs from Exoc5 control, KD, and OE MDCK cells, and subjected them mass spectrometry-based proteomic analysis to determine the protein composition of the EVs. By dendrogram, heat map, and principle component analysis, the protein content of the EVs was significantly different in Exoc5 KD, EXOC5 OE, and control MDCK cells. To confirm the proteomic results, we examined, by Western blot analysis, expression of the small guanosine triphosphate (GTP)-binding protein ADP-ribosylation factor (Arf6) and the epidermal growth factor receptor kinase substrate 8-like protein 2 (Eps8L2).

INSTRUMENT(S):

ORGANISM(S): Canis Familiaris (dog) (canis Lupus Familiaris)

TISSUE(S): Kidney

DISEASE(S): Polycystic Kidney Disease

SUBMITTER: Starr Hazard  

LAB HEAD: Josh Lipschutz, M.D.

PROVIDER: PXD013549 | Pride | 2019-11-06

REPOSITORIES: Pride