USP40 gene knockdown disrupts glomerular permeability in zebrafish

Hisashi Takagi, Yukino Nishibori, Kan Katayama, Tomohisa Katada, Shohei Takahashi, Zentaro Kiuchi, Shin Ichiro Takahashi, Hiroyasu Kamei, Hayato Kawakami, Yoshihiro Akimoto, Akihiko Kudo, Katsuhiko Asanuma, Hiromu Takematsu, Kunimasa Yan

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Abstract

Unbiased transcriptome profiling and functional genomics approaches have identified ubiquitin-specific protease 40 (USP40) as a highly specific glomerular transcript. This gene product remains uncharacterized, and its biological function is completely unknown. Here, we showed that mouse and rat glomeruli exhibit specific expression of the USP40 protein, which migrated at 150 kDa and was exclusively localized in the podocyte cytoplasm of the adult kidney. Double-labeling immunofluorescence staining and confocal microscopy analysis of fetal and neonate kidney samples revealed that USP40 was also expressed in the vasculature, including in glomerular endothelial cells at the premature stage. USP40 in cultured glomerular endothelial cells and podocytes was specifically localized to the intermediate filament protein nestin. In glomerular endothelial cells, immunoprecipitation confirmed actual protein-protein binding of USP40 with nestin, and USP40-small-interfering RNA transfection revealed significant reduction of nestin. In a rat model of minimal-change nephrotic syndrome, USP40 expression was apparently reduced, which was also associated with the reduction of nestin. Zebrafish morphants lacking Usp40 exhibited disorganized glomeruli with the reduction of the cell junction in the endothelium and foot process effacement in the podocytes. Permeability studies in these zebrafish morphants demonstrated a disruption of the selective glomerular permeability filter. These data indicate that USP40/Usp40 is a novel protein that might play a crucial role in glomerulogenesis and the glomerular integrity after birth through the modulation of intermediate filament protein homeostasis.

Original languageEnglish
Pages (from-to)F702-F715
JournalAmerican Journal of Physiology - Renal Physiology
Volume312
Issue number4
DOIs
Publication statusPublished - 04-2017

All Science Journal Classification (ASJC) codes

  • Physiology
  • Urology

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    Takagi, H., Nishibori, Y., Katayama, K., Katada, T., Takahashi, S., Kiuchi, Z., Takahashi, S. I., Kamei, H., Kawakami, H., Akimoto, Y., Kudo, A., Asanuma, K., Takematsu, H., & Yan, K. (2017). USP40 gene knockdown disrupts glomerular permeability in zebrafish. American Journal of Physiology - Renal Physiology, 312(4), F702-F715. https://doi.org/10.1152/ajprenal.00197.2016