Functional analysis of NPHS1 mutations in Japanese patients

Takayuki Miyai, Kunihiko Aya, Masanori Takaiwa, Kunimasa Yan, Yoshikazu Sado, Hiroyuki Tanaka, Tsuneo Morishima

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)279-284
Number of pages6
JournalHistology and Histopathology
Volume29
Issue number2
Publication statusPublished - Feb 2014
Externally publishedYes

Keywords

  • Congenital nephrotic syndrome
  • Nephrin
  • Podocin
  • Trafficking
  • Transient transfection

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

Fingerprint Dive into the research topics of 'Functional analysis of NPHS1 mutations in Japanese patients'. Together they form a unique fingerprint.

Cite this