RRC ID |
39514
|
著者 |
Tsuchida J, Matsusaka T, Ohtsuka M, Miura H, Okuno Y, Asanuma K, Nakagawa T, Yanagita M, Mori K.
|
タイトル |
Establishment of Nephrin Reporter Mice and Use for Chemical Screening.
|
ジャーナル |
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.
|
巻・号 |
11(6)
|
ページ |
e0157497
|
公開日 |
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
|
引用数 |
3
|
WOS 分野
|
UROLOGY & NEPHROLOGY
|
リソース情報 |
実験動物マウス |
RBRC01835 |