Real-time Imaging of Inositol 1,4,5-trisphosphate Movement in Mouse Salivary Gland Cells
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
Inositol 1,4,5-trisphosphate (IP₃) plays an important role in the release of Ca ²+ 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 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.
Abstract
Inositol 1,4,5-trisphosphate (IP₃) plays an important role in the release of Ca ²+ 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 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.
참고문헌 (27)
- Ashby MC, "Localized Ca2+ uncaging reveals polarized distribution of Ca2+-sensitive Ca2+ release sites: mechanism of unidirectional Ca2+ waves", JOURNAL OF CELL BIOLOGY, 158, pp.283~292 (2002)
- Berridge, MJ, "Calcium-a life and death signal", Nature, 395, pp.645~648 (1998)
- 정기명, "Cholinergic and neurokinergic agonist-induced Ca2+ responses in rat von Ebner's gland acinar cells", International Journal of Oral Biology, 31(3), pp.99~105 (2006), 대한구강생물학회
- Cocks, T.M, "A protective role for protease-activated receptors in the airways", Nature, 398(6723), pp.156~160 (1999)
- Czech MP, "PIP2 and PIP3: Complex Roles at the Cell Surface", Cell, 100, pp.603~606 (2000)
- Falasca M, "Activation of phospholipase Cγ by PI3-kinase-induced PH domain-mediated membrane targeting", EMBO Journal, 17, pp.414~422 (1998)
- Ferguson KM, "Crystal structure at 2.2 ?resolution of the pleckstrin homology domain from human dynamin", Cell, 79, pp.199~209 (1994)
- Fujii M, "Real-Time Visualization of PH Domain-Dependent Translocation of Phospholipase C-δ1 in Renal Epithelial Cells (MDCK): Response to Hypo-osmotic Stress", Biochemical and Biophysical Research Communications, 254, pp.284~291 (1999)
- Harlan JE, "Pleckstrin homology domains bind to phosphatidylinositol-4,5-bisphosphate", Nature, 371, pp.168~170 (1994)
- Hirose K, "Spatiotemporal Dynamics of Inositol 1,4,5-Trisphosphate That Underlies Complex Ca2+ Mob ilization Patterns", Science, 284, pp.1527~1530 (1999)
- Kawabata A, "The proteaseactivated receptor-2 agonist induces gastric mucus secretion and mucosal cytoprotection", Journal of Clinical Investigation, 107, pp.1443~1450 (2001)
- Kawabata A, "Proteaseactivated receptor-2 (PAR-2) in the pancreas and parotid gland: Immunolocalization and involvement of nitric oxide in the evoked amylase secretion", Life Sciences, 71, pp.2435~2446 (2002)
- Kawabata A, "Gastrointestinal functions of proteinase-activated receptors", Life Sciences, 74(2-3), pp.247~254 (2003)
- 김상희, "Purinergic Receptors Play Roles in Secretion of Rat von Ebner Salivary Gland", International Journal of Oral Biology, 31(4), pp.141~148 (2006), 대한구강생물학회
- Lemmon MA, "Specific and high-affinity binding of inositol phosphate to an isolated pleckstrin homology domain", Proceedings of the National Academy of Sciences of the United States of America, 92, pp.10472~10476 (1995)
- Matsu-ura T, "Cytosolic inositol 1,4,5-trisphosphate dynamics during intracellular calcium oscillations in living cells", Journal of Cell Biology, 173, pp.755~765 (2006)
- Mikoshiba K, "IP3 receptor", Ca2+ channel: from discovery to new signaling concepts, 2007, pp.102, 1426~1446
- Musacchio A, "The PH domain: a common piece in the structural pathcwork of signalling proteins", TRENDS IN BIOCHEMICAL SCIENCES, 18, pp.343~348 (1993)
- Nishikawa H, "Roles of Protease-Activated Receptor-2 (PAR-2), a G Protein-Coupled Receptor, in Modulation of Exocrine Gland Functions", Yakugaku Zassi, 126(7), pp.481~488 (2006)
- Payrastre Bdes, "Key players in cell signaling, in time and space", CELLULAR SIGNALLING, 13, pp.377~387 (2001)
- Rameh LE, "A comperative analysis of the phosphoinositide binding specificity of pleckstrin homology domains", Journal of Biological Chemistry, 272, pp.22059~22066 (1997)
- Sato M, "Locating inositol 1,4,5-trisphosphate in the nucleus and neuronal dendrites with genetically encoded fluorescent indicators", ANALYTICAL CHEMISTRY, 77, pp.4751~4758 (2005)
- i Waves in Polarized Exocrine Cells, "Journal of Biological Chemistry", 2001, 44146~44156
- Shin DM, "Homer 2 tunes G protein coupled receptors stimulus intensity by regulating RGS proteins and PLC β GAP activities", Journal of Cell Biology, 162, pp.293~303 (2003)
- Simonsen A, "The role of phosphoinositides in membrane transport", Current Opinion in Cell Biology, 13, pp.485~492 (2001)
- Tanimura A, "Fluorescent Biosensor for Quantitative Real-time Measurements of Inositol 1,4,5-Trisphosphate in Single Living Cells", Journal of Biological Chemistry, 279, pp.38095~38098 (2004)
- inositol-labeled phosphoinositide pools, "Journal of Cell Biology", 1998, 501~510