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Exosomal transfer from human renal proximal tubule cells to distal tubule and
collecting duct cells
#MMPMID24976626
Gildea JJ
; Seaton JE
; Victor KG
; Reyes CM
; Bigler Wang D
; Pettigrew AC
; Courtner CE
; Shah N
; Tran HT
; Van Sciver RE
; Carlson JM
; Felder RA
Clin Biochem
2014[Oct]; 47
(15
): 89-94
PMID24976626
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OBJECTIVES: Exosomes are 50-90nm extracellular membrane particles that may
mediate trans-cellular communication between cells and tissues. We have reported
that human urinary exosomes contain miRNA that are biomarkers for salt
sensitivity and inverse salt sensitivity of blood pressure. This study examines
exosomal transfer between cultured human renal proximal tubule cells (RPTCs) and
from RPTCs to human distal tubule and collecting duct cells. DESIGN AND METHODS:
For RPTC-to-RPTC exosomal transfer, we utilized 5 RPTC lines producing exosomes
that were fluorescently labeled with exosomal-specific markers CD63-EGFP or
CD9-RFP. Transfer between RPTCs was demonstrated by co-culturing CD63-EGFP and
CD9-RFP stable clones and performing live confocal microscopy. For RPTC-to-distal
segment exosomal transfer, we utilized 5 distal tubule and 3 collecting duct
immortalized cell lines. RESULTS: Time-lapse videos revealed unique proximal
tubule cellular uptake patterns for exosomes and eventual accumulation into the
multivesicular body. Using culture supernatant containing exosomes from 3 CD9-RFP
and 2 CD63-EGFP RPTC cell lines, all 5 distal tubule cell lines and all 3
collecting duct cell lines showed exosomal uptake as measured by microplate
fluorometry. Furthermore, we found that RPTCs stimulated with fenoldopam
(dopamine receptor agonist) had increased production of exosomes, which upon
transfer to distal tubule and collecting duct cells, reduced the basal reactive
oxygen species (ROS) production rates in those recipient cells. CONCLUSION: Due
to the complex diversity of exosomal contents, this proximal-to-distal vesicular
inter-nephron transfer may represent a previously unrecognized trans-renal
communication system.
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