| Project/Area Number |
13356008
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
ABE Keiko Graduate School of Agricultural and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (10151094)
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| Co-Investigator(Kenkyū-buntansha) |
SORIMACHI Hiroyuki Graduate School of Agricultural and Life Sciences, Associate professor, 大学院・農学生命科学研究科, 助教授 (10211327)
EMORI Yasufumi Graduate School of Science, Associate professor, 大学院・理学系研究科, 助教授 (60160389)
EBIHARA Satoshi National Cancer Center Hospital East, Director(Researcher), 院長(研究職)
TSUKANAOTO Yoshinori Mitsukan Group Corporation, Central Research Institute, Director(Researcher), 本社中央研究所, 所長(研究職)
NAKAO Masayuki Graduate School of Engineering, Associate professor, 大学院・工学系研究科, 助教授 (90242007)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥43,940,000 (Direct Cost: ¥33,800,000、Indirect Cost: ¥10,140,000)
Fiscal Year 2002: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
Fiscal Year 2001: ¥27,690,000 (Direct Cost: ¥21,300,000、Indirect Cost: ¥6,390,000)
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| Keywords | taste bud cell / primary culture / Ca ion / gene introduction / cell adhesion / basement membrane / laminin / integrin / 味覚 |
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| Research Abstract |
We have established an in vitro culture system of taste bud cells from rat circumvallate papillae. The taste bud cells were adhesive and viable for over 3 days when cultured onto Matrigel-coated dishes in medium based on keratinocyte growth medium. The cells retained molecular markers for both the cytoskeleton and intracellular signalling such as cytokeratin 8 and phospholipase Cβ2. In addition, three intracellular signaling molecules, gustducin, phospholipase Cβ2, and inositol 1, 4, 5-trisphosphate receptor type 3, are expressed in the same correlation as those in vivo. We show that an exogenously expressed myc-tagged α1 A-adrenoceptor sorts into the plasma membrane, and transduces a ligand-dependent signal resulting in intracellular [Ca^<2+>] increase in about half of the infected cells.
Under high Ca^<2+> condition (about 0.5 mM), the cells were tightly associated with each other and formed packed aggregates. Under low Ca^<2+> condition (below 1.1 mM), the cells were dispersed and had an elongated shape. These two forms were reversible and specifically dependent on Ca^<2+>. The results indicate that extracellular Ca^<2+> regulates cell shape and cell-to-cell adhesion of taste bud cells. We analyzed the subunits of laminin, a major component of the basement membrane, in circumvallate papillae of rat tongue by the method of RT-PCR and immunohistochemistry. As a result, the papillae contain β2 and γ1 as major β and γ subunits, respectively and three integrin β subunit species, β1, β4 and β5.
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