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KMID : 0848120080330040125
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International Journal of Oral Biology
2008 Volume.33 No. 4 p.125 ~ p.129
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Real-time Imaging of Inositol 1,4,5-trisphosphate Movement in Mouse
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Hong Jeong-Hee
Shin Dong-Min
Lee Syng-Ill
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Abstract
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Inositol 1,4,5-trisphosphate (IP3) plays an important role in the release of Ca2+ 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 (PLCδ1) 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 IP3 movement 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 IP3 intracellular dynamics.
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KEYWORD
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inositol 1, 4, 5-trisphosphate imaging, green fluorescent protein, transgenic mouse, parotid gland, submandibular gland
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