Expression of canonical transient receptor potential (TRPC) proteins in human glomerular mesangial cells

Sherry Sours, Juan Du, Shaoyou Chu, Min Ding, Xin J. Zhou, Rong Ma

Research output: Contribution to journalArticle

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Abstract

Mesangial cells are located within glomerular capillary loops and contribute to the physiological regulation of glomerular hemodynamics. The function of mesangial cells is controlled by a variety of ion channels in the plasma membrane, including nonselective cation channels, receptor-operated Ca2+ channels, and recently identified store-operated Ca2+ channels. Although the significance of these channels has been widely acknowledged, their molecular identities are still unknown. Recently, the members of the canonical transient receptor potential (TRPC) protein family have been demonstrated to behave as cation channels. The present study was performed to identify the isoforms of endogenous TRPC proteins in human mesangial cells (HMCs) and their interactions. Western blotting showed that TRPC1, 3, 4, and 6 were expressed in cultured HMCs. Consistently, immunofluorescent confocal microscopy revealed specific stainings for TRPC1, 3, 4, and 6 with predominant intracellular localization. However, TRPC5 and 7 were not detectable at protein level by either Western blotting or immunofluorescent staining. The expression of TRPC1, 3, 4, and 6 was also observed in rat and human glomeruli using fluorescent immunohistochemistry. Furthermore, coimmunoprecipitation experiments and immunofluorescent double staining displayed that TRPC1 had physical interaction with TRPC4 and 6, while no interactions were detected among other isoforms of TRPCs. Ca2+ fluorescent ratiometry measurement showed that store-operated Ca2+ entry in HMCs was significantly reduced by knocking down TRPC1, but enhanced by overexpressing TRPC1. These results suggest that HMCs specifically express isoforms of TRPC1, 3, 4, and 6 proteins. These isoforms of TRPCs might selectively assemble to form functional complexes.

Original languageEnglish
Pages (from-to)F1507-F1515
JournalAmerican Journal of Physiology - Renal Physiology
Volume290
Issue number6
DOIs
StatePublished - 1 Jun 2006

Fingerprint

Mesangial Cells
Protein Isoforms
Proteins
Staining and Labeling
Cations
Western Blotting
Ion Channels
Confocal Microscopy
Cell Communication
Hemodynamics
Immunohistochemistry
Cell Membrane

Keywords

  • Ca channel
  • Ca signaling
  • Glomeruli
  • Store-operated ca entry

Cite this

Sours, Sherry ; Du, Juan ; Chu, Shaoyou ; Ding, Min ; Zhou, Xin J. ; Ma, Rong. / Expression of canonical transient receptor potential (TRPC) proteins in human glomerular mesangial cells. In: American Journal of Physiology - Renal Physiology. 2006 ; Vol. 290, No. 6. pp. F1507-F1515.
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Expression of canonical transient receptor potential (TRPC) proteins in human glomerular mesangial cells. / Sours, Sherry; Du, Juan; Chu, Shaoyou; Ding, Min; Zhou, Xin J.; Ma, Rong.

In: American Journal of Physiology - Renal Physiology, Vol. 290, No. 6, 01.06.2006, p. F1507-F1515.

Research output: Contribution to journalArticle

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AU - Sours, Sherry

AU - Du, Juan

AU - Chu, Shaoyou

AU - Ding, Min

AU - Zhou, Xin J.

AU - Ma, Rong

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AB - Mesangial cells are located within glomerular capillary loops and contribute to the physiological regulation of glomerular hemodynamics. The function of mesangial cells is controlled by a variety of ion channels in the plasma membrane, including nonselective cation channels, receptor-operated Ca2+ channels, and recently identified store-operated Ca2+ channels. Although the significance of these channels has been widely acknowledged, their molecular identities are still unknown. Recently, the members of the canonical transient receptor potential (TRPC) protein family have been demonstrated to behave as cation channels. The present study was performed to identify the isoforms of endogenous TRPC proteins in human mesangial cells (HMCs) and their interactions. Western blotting showed that TRPC1, 3, 4, and 6 were expressed in cultured HMCs. Consistently, immunofluorescent confocal microscopy revealed specific stainings for TRPC1, 3, 4, and 6 with predominant intracellular localization. However, TRPC5 and 7 were not detectable at protein level by either Western blotting or immunofluorescent staining. The expression of TRPC1, 3, 4, and 6 was also observed in rat and human glomeruli using fluorescent immunohistochemistry. Furthermore, coimmunoprecipitation experiments and immunofluorescent double staining displayed that TRPC1 had physical interaction with TRPC4 and 6, while no interactions were detected among other isoforms of TRPCs. Ca2+ fluorescent ratiometry measurement showed that store-operated Ca2+ entry in HMCs was significantly reduced by knocking down TRPC1, but enhanced by overexpressing TRPC1. These results suggest that HMCs specifically express isoforms of TRPC1, 3, 4, and 6 proteins. These isoforms of TRPCs might selectively assemble to form functional complexes.

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