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Open AccessArticle

Urinary Extracellular Vesicles and Salt-Losing Tubulopathies: A Proteomic Approach

1
School of Medicine and Surgery, University Milan Bicocca, 20900 Monza, Italy
2
Laboratory of Medical Genetics, Fondazione IRCCS Ca’ Granda Hospital, 20162 Milan, Italy
*
Author to whom correspondence should be addressed.
Proteomes 2020, 8(2), 9; https://doi.org/10.3390/proteomes8020009
Received: 28 February 2020 / Revised: 17 April 2020 / Accepted: 8 May 2020 / Published: 9 May 2020
(This article belongs to the Special Issue Human Proteomics)
Renal tubular cells release urinary extracellular vesicles (uEV) that are considered a promising source of molecular markers for renal dysfunction and injury. We investigated uEV proteomes of patients with hereditary salt-losing tubulopathies (SLTs), focusing on those caused by Gitelman and Bartter (BS) syndromes, to provide potential markers for differential diagnosis. Second morning urine was collected from patients with genetically proven SLTs and uEV were isolated by the ultracentrifugation-based protocol. The uEV proteome was run through a diagonal bidimensional electrophoresis (16BAC/SDS-PAGE), to improve hydrophobic protein resolution. Sixteen differential spots from the proteome of two variants (BS2 and BS3) were analysed by nLC-ESI-MS/MS after in-gel tryptic digestion. A total of 167 protein species were identified from 7 BS2 spots and 9 BS3 spot. Most of these proteins were membrane-associated proteins, in particular transmembrane proteins, and were related to typical renal functions. The differential content of some uEV was then validated by immunoblotting. Our work suggests that uEV proteomics represents a promising strategy for the identification of differential SLT proteins. This could play a role in understanding the pathophysiological disease mechanisms and may support the recognition of different syndromes. View Full-Text
Keywords: exosomes; extracellular vesicles; Gitelman syndrome; Bartter syndrome; diagonal electrophoresis exosomes; extracellular vesicles; Gitelman syndrome; Bartter syndrome; diagonal electrophoresis
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MDPI and ACS Style

Raimondo, F.; Chinello, C.; Porcaro, L.; Magni, F.; Pitto, M. Urinary Extracellular Vesicles and Salt-Losing Tubulopathies: A Proteomic Approach. Proteomes 2020, 8, 9.

AMA Style

Raimondo F, Chinello C, Porcaro L, Magni F, Pitto M. Urinary Extracellular Vesicles and Salt-Losing Tubulopathies: A Proteomic Approach. Proteomes. 2020; 8(2):9.

Chicago/Turabian Style

Raimondo, Francesca; Chinello, Clizia; Porcaro, Luigi; Magni, Fulvio; Pitto, Marina. 2020. "Urinary Extracellular Vesicles and Salt-Losing Tubulopathies: A Proteomic Approach" Proteomes 8, no. 2: 9.

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