| Project/Area Number |
09557008
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
General pharmacology
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| Research Institution | Yamagata University |
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Principal Investigator |
KATANO Yumi Yamagata University School of Medicine, Professor, 医学部, 教授 (70018696)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Eiji Yamagata University School of Medicine, Associate Professor, 医学部, 助教授 (30206792)
ISHIHATA Akira Yamagata University School of Medicine, Instructor, 医学部, 助手 (40232326)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1997: ¥6,300,000 (Direct Cost: ¥6,300,000)
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| Keywords | aging-associated genes / cloning / myocardium / differential display / rat / Northern blot / unknown genes / gene expression / 老化関連遺伝子 / differential display / ノーザンブロット解析 / RT-PCR / アドレノメデュリン / Coronary flow / NO / PGI_2 / 心機能 / アドレノメデュリン受容体遺伝子 / Aging / エンドセリン-1 / エンドセリン受容体 / エンドセリン受容体遺伝子 / ラット心筋 / 陽性変力作用 / 筋小胞体Ca^<++>取り込み / 加齢変化 / ノーザンブロッティング |
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| Research Abstract |
It has been known that the physiological function of the heart is altered according to the aging. For example, the positive inotropic effects of isoproterenol and endothelin-1 are decreased by aging in the rat ventricles. While the binding capacity for b-adrenergic receptor was decreased, the specific binding capacity of endothelin-1 was increased in the aged rats. In sarcoplasmic reticulum, the activity of CaィイD12+ィエD1 was reduced. In coronary arterial endothelium, nitric oxide production was very low-in aged rat. The molecular basis of these age-dependent functional changes has not been understood. Differential gene expression provides the basis for many fundamental cellular processes associated with differentiation, development and aging. The identification and cloning of genes whose expression is modulated is an important step in advancing our understanding of aging phenomena. In order to isolate and characterize gene products differentially expressed in aging, we applied the diffe
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rential display reverse transcription-polymerase chain reaction for screening those genes in 2-3 months-old and 27 months-old Fischer 344 rat ventricles. Total RNAs were isolated by acid-guanidium-phenol-chloroform method. Messenger RNAs were obtained from total RNA by using oligo d(T) column. The first-strand cDNAs from each RNA were synthesized with three oligo-d(T)-anchor primers. PCR was carried out with anchor primers and eight arbitrary primers, and the products were analyzed on sequence gels. The 19 unique PCR products displayed increased expression in aged heart, while 66 clones were decreased in aged heart. Differential expression was confirmed by Northern blot analysis or reverse Northern blotting. The clones included genes for mitochondrial metabolism such as cytochrome c oxidase, myosin heavy chain, cell surface glycoproteins and several kinases/phosphatases for signal transduction. 31 clones were sequenced and 14 clones were unknown genes. The method used in this study is valuable for studying the age-associated gene expression, and we are continuing to analyze the unknown genes identified in this study. Less
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