Phospholipase C-γ1 is required for the activation of store-operated Ca2+ channels in liver cells

Tom Litjens, Than Nguyen, Joel Castro, Edoardo C. Aromataris, Lynette Jones, Greg J. Barritt, Grigori Y. Rychkov

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31 Citations (Scopus)

Abstract

Repetitive hormone-induced changes in concentration of free cytoplasmic Ca2+ in hepatocytes require Ca2+ entry through receptor-activated Ca2+ channels and SOCs (store-operated Ca 2+ channels). SOCs are activated by a decrease in Ca2+ concentration in the intracellular Ca2+ stores, but the molecular components and mechanisms are not well understood. Some studies with other cell types suggest that PLC-γ (phospholipase C-γ) is involved in the activation of receptor-activated Ca2+ channels and/or SOCs, independently of PLC-γ-mediated generation of IP3 (inositol 1,4,5-trisphosphate). The nature of the Ca2+ channels regulated by PLC-γ has not been defined clearly. The aim of the present study was to determine if PLC-γ is required for the activation of SOCs in liver cells. Transfection of H4IIE cells derived from rat hepatocytes with siRNA (short interfering RNA) targeted to PLC-γ1 caused a reduction (by approx. 70%) in the PLC-γ1 protein expression, with maximal effect at 72-96 h. This was associated with a decrease (by approx. 60%) in the amplitude of the I SOC (store-operated Ca2+ current) developed in response to intracellular perfusion with either IP3 or thapsigargin. Knockdown of STIM1 (stromal interaction molecule type 1) by siRNA also resulted in a significant reduction (approx. 80 % at 72 h post-transfection) of the I SOC amplitude. Immunoprecipitation of PLC-γ1 and STIM1, however, suggested that under the experimental conditions these proteins do not interact with each other. It is concluded that the PLC-γ1 protein, independently of IP3 generation and STIM1, is required to couple endoplasmic reticulum Ca2+ release to the activation of SOCs in the plasma membrane of H4IIE liver cells.

Original languageEnglish
Pages (from-to)269-276
Number of pages8
JournalBiochemical Journal
Volume405
Issue number2
DOIs
Publication statusPublished - 15 Jul 2007

Keywords

  • Phospholipase C (PLC)
  • Short interfering RNA (siRNA)
  • Store-operated Ca channel (SOC)
  • Stromal interaction molecule type 1 (STIM1)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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