ISSN : 1226-7155(Print)
ISSN : 2287-6618(Online)
ISSN : 2287-6618(Online)
Real-time Imaging of Inositol 1,4,5-trisphosphate Movement in Mouse Salivary Gland Cells
Abstract
Inositol 1,4,5-trisphosphate (IP₃) plays an important role in the release of Cα²+ from intracellular stores into the cytoplasm in a variety of cell types. IP₃ translocation dynamics have been studied in response to many types of cell signals. However, the dynamics of cytosolic IP₃ in salivary acinar cells are unclear. A green fluorescent protein (GFP)-tagged pleckstrin homology domain (PHD) was constructed and introduced into a phospholipase C δ1 (PLC δ1) transgenic mouse, and then the salivary acinar cells were isolated. GFP-PHD was heterogeneously localized at the plasma membrane and intracellular organelles in submandibular gland and parotid gland cells. Application of trypsin, a G protein-coupled receptor activator, to the two types of cells caused an increase in GFP fluorescence in the cell cytoplasm. The observed time course of trypsin-evoked IP₃movement in acinar cells was independent of cell polarity, and the fluorescent label showed an immediate increase throughout the cells. These results suggest that GFP-PHD in many tissues of transgenic mice, including non-cultured primary cells, can be used as a model for examination of IP₃intracellular dynamics.
inositol 1,4,5-trisphosphate imaging; green fluorescent protein; transgenic mouse; parotid gland; submandibular gland
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