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J Proteomics. 2014 Apr 4;100:125-35. doi: 10.1016/j.jprot.2013.11.025. Epub 2013 Dec 4.

Proteomic analysis of rat proximal tubule cells following stretch-induced apoptosis in an in vitro model of kidney obstruction.

Author information

1
Department of Pathology, 11th Floor Tupper Medical Building, Room 11B, B3H 4R2, Dalhousie University, Halifax, NS, Canada. Electronic address: Dennis.orton@dal.ca.
2
Department of Chemistry, Room 212, Chemistry Building, B3H 4R2, Dalhousie University, Halifax, NS, Canada. Electronic address: Alan.doucette@dal.ca.
3
Department of Biomedical Engineering, 4th Floor Dentistry Building, B3H 1 W2, Dalhousie University, Halifax, NS, Canada. Electronic address: Gmaksym@dal.ca.
4
Department of Pathology, 11th Floor Tupper Medical Building, Room 11B, B3H 4R2, Dalhousie University, Halifax, NS, Canada. Electronic address: Dawn.MacLellan@iwk.nshealth.ca.

Abstract

Urinary tract obstruction (UTO) is a commonly noted disorder on prenatal ultrasound that has the potential to lead to permanent loss of renal function. To study the molecular processes of the disease, an in vitro model has been developed which involves mechanical stretch of proximal tubule cells grown on flexible plates which mimics the physiological conditions during UTO. This study employs a one dimensional SDS-PAGE fractionation procedure, followed by in-gel digest and LC-MS/MS analysis in a semi-quantitative experiment using spectral counting to relatively quantify changes in protein expression following the established model of UTO. Quantitative analysis shows 317 of the 1630 identified proteins express altered abundance, with 135 increased and 182 decreased in abundance as a result of stretch. Gene ontology (GO) and KEGG annotations implicate a number of physiological processes that have been previously shown in addition to some potentially novel processes in UTO. The quantitative proteomic analysis we performed here provides a more complete characterization of changes in protein abundance as a result of stretch than previous studies, and provides a number of previously undescribed proteins in proximal tubule cells that may play a role in UTO.

BIOLOGICAL SIGNIFICANCE:

Urinary tract obstruction (UTO) is a commonly noted abnormality on prenatal ultrasound that can either resolve spontaneously or require surgical intervention to prevent permanent renal damage or loss of function. While targeted studies of UTO have shown a number of pathological responses in proximal tubule cells, there are currently no large-scale quantitative studies that aim to elucidate a global cellular response. This study uses a semi-quantitative approach and applies a well characterized model of UTO to determine a number of cellular processes affected by UTO simulation and identifies a number of proteins with altered abundance that have not been noted previously in UTO. This article is part of a Special Issue entitled: Can Proteomics Fill the Gap Between Genomics and Phenotypes?

KEYWORDS:

In-gel digest; Proximal tubule; Spectral counting; Urinary tract obstruction

PMID:
24316357
DOI:
10.1016/j.jprot.2013.11.025
[Indexed for MEDLINE]

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