1995 Fiscal Year Final Research Report Summary
Analysis of Phenotypic Variation of Mast Cells by Using Messenger RNA Expression as a Marker
| Project/Area Number |
06454193
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | Osaka University |
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Principal Investigator |
KITAMURA Yukihiko Dept.Path.Prof., 医学部, 教授 (70028520)
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| Co-Investigator(Kenkyū-buntansha) |
MORII Eiichi Dept.Path.Instruct., 医学部, 特別研究員
TSHUJIMRA Tohru Dept.Path.Instruct., 医学部, 助手 (20227408)
HIROTA Seiichi Dept.Path.Instruct., 医学部, 助手 (50218856)
NOMURA Shintaro Dept.Path.Assoc.Prof, 医学部, 助教授 (80159087)
JIPPO Tomoko Dept.Path.Instruct., 医学部, 助手 (70252658)
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| Project Period (FY) |
1994 – 1995
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| Keywords | mast cell / protease / in situ hybridization / cytokine / cytokine receptor / transcription factor / tryptase / chymase |
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| Research Abstract |
Basophilic granules of mase cells contain various mast cell-specific proteases. Many genes encoding mast cell-specific proteases have been cloned in mice. Mouse mast cells proteases (MMCP) 1 to 5 are chymases, and MMCP-6 and -7 are tryptases. We investigated the factors which influenced the expression of mast cell-specific proteases by using in situ hybridization. The pattern of MMCP expression was affected by mouse atrains, tissues in which mast cells differentiated and signals from cytokine receptors. In 1994, we obtained cultured mast cells (CMCs) by cultureing bone marrow cells of mice in the presecnce of T cell-derived cytokines. Total RNA was extracted from CMCs, and cDNAs of MMCP-2, MMCP-4, MMCP-5, MMCP-6 and mast cell carboxypeptidase were obtained. CMCs were centrifuged and embedded in paraffin, and in situ hybridiazation using digoxigenin-labeled RNA probes was carried out. Since the technical problems were resolved in 1994, we studid the change in mRNA expression patters of mast cell-specific proteases under various culture condition in 1995. We found the gain-of-function mutations of c-kit receptor tyrosine kinase, which is the receptor for stem cell factor, the most important growth factor of mast cells. We investifated the effect of signals through the c-kit receptor on the expression of mast cell-specific proteases. Introduction of the constitutively activated c-kit cDNA into the IC-2 mast cell line resulted in the enhanced expression of MMCP-6.
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