Decreasing calreticulin expression lowers the Ca2+ response to bradykinin and increases sensitivity to ionomycin in NG-108-15 cells.

It has been suggested that the multifunctional protein, calreticulin, is a major calcium sequestering protein in the inositol 1,4,5-trisphosphate receptor-containing endoplasmic reticulum subcompartment. In neuroblastoma X glioma NG-108-15 cells, bradykinin can effectively stimulate the release of inositol 1,4,5-trisphosphate and cause a cytosolic calcium transient. To explore the function of calreticulin as an intracellular calcium sequestering protein, we investigated calcium dynamics in NG-108-15 cells after treatment with an antisense oligonucleotide against calreticulin, CrtAS1. Cells treated with either CrtAS1 or the corresponding sense oligonucleotide CrtPS1 were examined for their calreticulin content by Western blotting, the amplitude of their calcium transient in response to bradykinin, and their sensitivity toward the calcium ionophore, ionomycin. Treatment with CrtAS1 decreased the amount of calreticulin in comparison to CrtPS1-treated and untreated control cells. At the same time, CrtAS1-treated cells had a significantly reduced calcium response to bradykinin and were more sensitive to ionomycin-induced cell death. These data show that the level of calreticulin expression is directly related to the calcium storage capacity of the inositol 1,4,5-trisphosphate-sensitive calcium pool and indicate a direct relationship between the level of calreticulin and the protection against cytotoxic calcium overload.