Project/Area Number |
11306013
|
Research Category |
Grant-in-Aid for Scientific Research (A)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Fisheries chemistry
|
Research Institution | THE UNIVERSITY OF TOKYO |
Principal Investigator |
WATABE Shugo Graduate School of Agricultural and Life Sciences, The University Of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (40111489)
|
Co-Investigator(Kenkyū-buntansha) |
SAKAI Tadashi Miyazaki University, Faculty of Agriculture, Professor, 農学部, 教授 (60136794)
USHIO Hideki Tokyo University of Fisheries, Faculty of Fisheries, Associate Professor, 水産学部, 助教授 (50251682)
KOBAYASHI Makito Graduate School of Agricultural and Life Sciences, The University Of Tokyo, Professor, 大学院・農学生命科学研究科, 助教授 (30183809)
HIRAYAMA Yasushi Japan Society for the Promotion of Science, Postdoctoral Researcher, 特別研究員
|
Project Period (FY) |
1999 – 2001
|
Project Status |
Completed (Fiscal Year 2001)
|
Budget Amount *help |
¥40,470,000 (Direct Cost: ¥38,400,000、Indirect Cost: ¥2,070,000)
Fiscal Year 2001: ¥8,970,000 (Direct Cost: ¥6,900,000、Indirect Cost: ¥2,070,000)
Fiscal Year 2000: ¥10,200,000 (Direct Cost: ¥10,200,000)
Fiscal Year 1999: ¥21,300,000 (Direct Cost: ¥21,300,000)
|
Keywords | Temperature adaptation / Fish / Mg^<2+>-ATPase activity / Myosin heavy Chain gene / Mitochondria / Membrane voltage potential / Myofiril / 5' upstream region / Mg^<2+>ATPase活性 / 細胞小器官 / チロキシン / 高温耐性ニジマス / 筋小胞体 / ホルモン調節 / 遺伝子調節 / 分子機構 / ミオシン |
Research Abstract |
Carp acclimates to environmental temperature and induces genes encoding myosin heavy chain isoforms predominantly expressed for fish acclimated to 10 and 30 ℃. Such changes in biochemical changes in carp following temperature acclimation are also observed with intracellular apparatus such as sarcoplasmic reticulum and various enzymes. It seems that, fish alters dynamically many cellular components in response to environmental temperatures. It is considered that environmental temperature signals are transmitted to the central nervous system and subsequently act on gene expression via various hormonal regulations. The objective of this study was to disclose molecular mechanisms involved in temperature adaptation of fish and to utilize the results obtained for industrial application. The 5' upstream regulatory regions of the 10 ℃- and 30 ℃-type carp myosin heavy chain isoforms were cloned and determined for their structural integrity, resulting in identification of MEF2 binding element res
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ponsible for activation of gene expression. Furthermore, it was found that 5'-upstream regions of myosin heavy chain genes from medaka, another eurythermal fish, also participate in such regulation for temperature-dependent gene expression. Carp increased expression levels of 55 kDa cytosolic protein in association with acclimation to cold temperature. This protein was found to be one of subunits of mitochondrial FOF1-ATPase and increased its quantities two times following temperature shift from 30 ℃ to 10 ℃. Such different expression depending on acclimation temperature was regulated at the transcription level. Various fish species were collected from Kasumigaura Lake periodically all around the year and determined for ATPase activity of fast skeletal muscle myofibrils. The activity was the highest with fish collected in November and decreased gradually reaching the minimum value in June and July. Analytical method to determine mitochondrial membrane voltage was developed by using voltage-sensitive probe and fluorescence imaging. Changes in voltage-associated potential energy of mitochondrial membranes following adaptation to environmental temperatures of fish were successfully measured by this method. Less
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