osr1 maintains renal progenitors and regulates podocyte development by promoting wnt2ba through antagonism of hand2

ABSTRACTKnowledge about the genetic pathways that control renal cell lineage development is essential to better understand the basis of congenital malformations of the kidney and design regenerative medicine therapies. The embryonic zebrafish kidney, or pronephros, contains two nephrons that are conserved with humans. Recently, the transcription factors Osr1 and Hand2 were found to exert antagonistic influences to balance kidney specification (Perens et al., 2016). Here, we performed a forward genetic screen in zebrafish to identify nephrogenesis regulators, where whole genome sequencing of the novel oceanside (ocn) mutant revealed a nonsense mutation in osr1. ocn mutants evince severe pronephros defects including abrogation of podocytes and proximal tubule cells. Our studies reveal that osr1 is not needed to specify renal progenitors, but rather required to maintain their survival. Additionally, osr1 is requisite for expression of the canonical Wnt ligand wnt2ba, where wnt2ba is expressed in the intermediate mesoderm (IM) and later restricts to podocytes. Deficiency of wnt2ba reduced podocyte progenitors, where overexpression of wnt2ba was sufficient to rescue the podocyte lineage as well as osr1 loss of function. Finally, we demonstrate that reciprocal antagonism between osr1 and hand2 mediates podocyte development specifically by controlling wnt2ba expression in the IM. Together, our data show that Osr1 is essential for a sequence of temporal functions that mediate the survival and lineage decisions of IM progenitors, and subsequently the maintenance of podocytes and proximal tubule epithelium in the embryonic nephron.

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Toxicoproteomic Analysis of Poly(ADP-Ribose)-Associated Proteins Induced by Oxidative Stress in Human Proximal Tubule Cells

Toxicological Sciences ◽

10.1093/toxsci/kfz131 ◽

2019 ◽

Vol 171 (1) ◽

pp. 117-131

Author(s):

Argel Islas-Robles ◽

Deepthi Yedlapudi ◽

Serrine S Lau ◽

Terrence J Monks

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Proximal Tubule ◽

Calcium Influx ◽

Regulation Of Transcription ◽

Loss Of Function ◽

Proximal Tubule Cells ◽

Associated Proteins ◽

Tubule Cells ◽

Parp 1

Abstract 2,3,5-Tris-(glutathion-S-yl)hydroquinone (TGHQ) is a nephrotoxic and nephrocarcinogenic metabolite of hydroquinone. TGHQ generates reactive oxygen species (ROS), causing DNA-strand breaks, hyperactivation of PARP-1, increases in intracellular calcium ([Ca2+]i), and cell death. PARP-1 catalyzes the attachment of ADP-ribose polymers (PAR) to target proteins. In human kidney proximal tubule cells, ROS-mediated PARP-1 hyperactivation and elevations in [Ca2+]i are reciprocally coupled. The molecular mechanism of this interaction is unclear. The aim of the present study was to identify ROS-induced PAR-associated proteins to further understand their potential role in cell death. PAR-associated proteins were enriched by immunoprecipitation, identified by LC-MS/MS, and relative abundance was obtained by spectral counting. A total of 356 proteins were PAR-modified following TGHQ treatment. A total of 13 proteins exhibited gene ontology annotations related to calcium. Among these proteins, the general transcription factor II-I (TFII-I) is directly involved in the modulation of [Ca2+]i. TFII-I binding to phospholipase C (PLC) leads to calcium influx via the TRPC3 channel. However, inhibition of TRPC3 or PLC had no effect on TGHQ-mediated cell death, suggesting that their loss of function may be necessary but insufficient to cause cell death. Nevertheless, TGHQ promoted a time-dependent translocation of TFII-I from the nucleus to the cytosol concomitant with a decrease in tyrosine phosphorylation in α/β-TFII-I. Therefore it is likely that ROS have an important impact on the function of TFII-I, such as regulation of transcription, and DNA translesion synthesis. Our data also shed light on PAR-mediated signaling during oxidative stress, and contributes to the development of strategies to prevent PAR-dependent cell death.

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Acute Kidney Injury in SARS-CoV-2 Infection: Direct Effect of Virus on Kidney Proximal Tubule Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21093275 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3275 ◽

Cited By ~ 10

Author(s):

Manoocher Soleimani

Keyword(s):

Acute Kidney Injury ◽

Severe Acute Respiratory Syndrome ◽

Proximal Tubule ◽

Rna Viruses ◽

Kidney Injury ◽

The Novel ◽

Proximal Tubule Cells ◽

Severe Respiratory Infection ◽

Tubule Cells ◽

Novel Coronavirus

Coronaviruses (CoVs), including Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), and the novel coronavirus disease-2 (SARS-CoV-2) are a group of enveloped RNA viruses that cause a severe respiratory infection which is associated with a high mortality [...]

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Faculty Opinions recommendation of Mitochondrial aquaporin-8 in renal proximal tubule cells: evidence for a role in the response to metabolic acidosis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717948404.793453455 ◽

2012 ◽

Author(s):

Jeff Kraut

Keyword(s):

Metabolic Acidosis ◽

Proximal Tubule ◽

Renal Proximal Tubule ◽

Proximal Tubule Cells ◽

Tubule Cells ◽

Renal Proximal Tubule Cells

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Nephrotoxicity Testing In Vitro: The Current Situation

Alternatives to Laboratory Animals ◽

10.1177/026119299702500506 ◽

1997 ◽

Vol 25 (5) ◽

pp. 497-503

Author(s):

Jean-Paul Morin ◽

Marc E. De Broe ◽

Walter Pfaller ◽

Gabriele Schmuck

Keyword(s):

Proximal Tubule ◽

Culture Media ◽

Task Force ◽

Primary Cultures ◽

Phenotypic Expression ◽

Transepithelial Transport ◽

Specific Cell ◽

Proximal Tubule Cells ◽

Tubule Cells

An ECVAM task force on nephrotoxicity has been established to advise, in particular, on the follow-up to recommendations made in the ECVAM workshop report on nephrotoxicity testing in vitro. Since this workshop was held, in 1994, there have been several improvements in the techniques used. For example, the duration of renal slice viability, and the maintenance of functional activities in slices, have been improved by using dynamic incubation systems with higher oxygen tensions and more-appropriate cell culture media. Highly differentiated primary cultures of pig, human and rabbit proximal tubule cells have been established by using specific cell isolation procedures and/or selective culture media. To date, the most comparable phenotypic expression and transepithelial transport capacities to proximal tubules in vivo have been obtained with primary cultures of rabbit proximal tubule cells which are grown on bicompartmental supports; in this system, transepithelial substrate gradients are generated and the transepithelial transport of both organic anions and cations is highly active. This in vitro system has been selected by ECVAM for further evaluation and prevalidation. Industrial needs in the area of nephrotoxicity testing have been identified, and recommendations are made at the end of this report concerning possible future initiatives.

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