1989 Fiscal Year Final Research Report Summary
New Marker of Thyroid Hormone Action: Identification of Thyroid Hormone-sensitive Gene in Human Lymphocytes.
| Project/Area Number |
63570555
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | Okinaka Memorial Institute for Medical Research |
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Principal Investigator |
OZAWA Yasunori Okinaka Memorial Institute for Medical Research. Senior Researcher., 主任研究員 (10124306)
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| Co-Investigator(Kenkyū-buntansha) |
ODAJIMA Ritsuko Okinaka Memorial Institute for Medical Research. Researcher., 研究員
SHISHIBA Yoshimasa Okinaka Memorial Institute for Medical Research. Senior Researcher., 主任研究員 (00072596)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Thyroid hormone sensitive gene / Human lymphocyte / Human skin fibroblast / Thyroid hormone conjugate |
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| Research Abstract |
In order to get precise and specific markers of biological action of thyroid hormones in human, we tried to identify genes and gene products which are controlled by thyroid hormone. We first tried to get mRNA of T3-sensitive genes in lymphcytes. Peripheral lymphocytes (B and T cells) were incubated with various amount of T3, and the mRNA was extracted from the cells thus incubated. Before selection hybridization, polyA-RNA was applied to cell free system, and resulting products was analyzed by two dimensional electrophoresis. Some spots on electrophoresis appeared to be increased by T3, but with poor reproducibility. The increments of the amount of mRNA which seemed reproducible between experiments were not enough high for further analysis. Additionally, total amount of polyA-RNA extracted from lymphocyte was not much enough. The reasons of these disappointing results seemed to be heterogeneity of cell population and cell-cell interaction during the incubation. Then, we tried to study T3-sensitive gene and gene products in human skin fibroblasts. We established a human skin fibroblast cell fine from a normal individual. This cell line had T3 nuclear receptors. Translation products of polyA-RNA extracted from these ceils incubated with or without T3 were applied for two dimensional electrophoresis. Six spots showed constant increase by T3. Now we are trying selection hybridization. Additionally, we verified the formation and secretion of the conjugates of thyroid hormone in intact cell before the interaction of thyroid hormone and receptor. Furthermore, we clarified that intracellular metabolites of sugar affect the thyroid hormone sensitive gene, Spot 14. This finding indicates other factors than thyroid hormone can affect thyroid hormone sensitive genes.
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