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ID 52739
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Author
Miyai, Takayuki
Aya, Kunihiko
Takaiwa, Masanori
Yan, Kunimasa
Sado, Yoshikazu
Tanaka, Hiroyuki
Morishima, Tsuneo
Abstract
Background: Many mutations in the NPHS1 gene were detected among patients with congenital nephrotic syndrome. Functional analysis of those mutations was done with a stable-expression cell line. Nevertheless, establishing such a cell line is time-consuming. Methods and Results: We established an easier method using automatic counting software for functional analysis with transient-transfection cells rather than a stable-expression cell line. We demonstrated maltrafficking to the plasma membrane of abnormal nephrin for immunostaining on transient-expression cells by comparison without Triton X (detecting proteins on the cell membrane only) and with Triton X (detecting proteins both on the cell membrane and inside the cell cytoplasm). We obtained relevant results with data obtained previously using a stable-expression cell line. Furthermore, we conducted functional analysis of NPHS1 mutations in Japanese patients with congenital nephrotic syndrome using this simple method, which revealed that all pathogenic mutations impaired trafficking to the protein plasma membrane. Conclusions: Functional analysis using transient-expression cells with automatic counting software was useful to demonstrate maltrafficking to the plasma membrane of a protein. All pathogenic mutations detected in Japanese patients impaired trafficking to the protein plasma membrane.
Keywords
Congenital nephrotic syndrome
Nephrin
Podocin
Trafficking
Transient transfection
Published Date
2014-02
Publication Title
Histology and Histopathology
Volume
volume29
Issue
issue2
Start Page
279
End Page
284
ISSN
0213-3911
Content Type
Journal Article
Official Url
http://www.hh.um.es/Abstracts/Vol_29/29_2/29_2_279.htm
Related Url
http://ousar.lib.okayama-u.ac.jp/metadata/52505
language
英語
Copyright Holders
© Servicio de Publicaciones, Universidad de Murcia, 2013
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Refereed
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