RRC ID 39514
Author Tsuchida J, Matsusaka T, Ohtsuka M, Miura H, Okuno Y, Asanuma K, Nakagawa T, Yanagita M, Mori K.
Title Establishment of Nephrin Reporter Mice and Use for Chemical Screening.
Journal PLoS One
Abstract Nephrin is a critical component of glomerular filtration barrier, which is important to maintain glomerular structure and avoid proteinuria. Downregulation of nephrin expression is commonly observed at early stage of glomerular disorders, suggesting that methods to increase nephrin expression in podocytes may have therapeutic utility. Here, we generated a knockin mouse line carrying single copy of 5.5 kb nephrin promoter controlling expression of enhanced green fluorescent protein (EGFP) at Rosa26 genomic locus (Nephrin-EGFP mouse). In these mice, EGFP was specifically expressed in podocytes. Next, we isolated and cultivated glomeruli from these mice, and developed a protocol to automatically quantitate EGFP expression in cultured glomeruli. EGFP signal was markedly reduced after 5 days of culture but reduction was inhibited by vitamin D treatment. We confirmed that vitamin D increased mRNA and protein expression of endogenous nephrin in cultivated glomeruli. Thus, we generated a mouse line converting nephrin promoter activity into fluorescence, which can be used to screen compounds having activity to enhance nephrin gene expression.
Volume 11(6)
Pages e0157497
Published 2016-1-1
DOI 10.1371/journal.pone.0157497
PII PONE-D-16-02984
PMID 27362433
PMC PMC4928931
MeSH Animals Cells, Cultured Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Kidney Glomerulus / metabolism* Membrane Proteins / genetics Membrane Proteins / metabolism* Mice Mice, Transgenic Podocytes / metabolism* Promoter Regions, Genetic
IF 2.74
Times Cited 3
Mice RBRC01835