Temperature adaptation of fish - molecular mechanisms involved and application

鱼类的温度适应——涉及的分子机制及应用

基本信息

批准号：
11306013
负责人：
WATABE Shugo
金额：
$ 25.9万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

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 … More 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

鲤鱼适应环境温度，并诱导编码肌球蛋白重链同工型的基因，主要表示适应于10和30°的鱼。还观察到温度适应后鲤鱼的生化变化的这种变化，也可以使用细胞内设备（例如肌浆网和各种酶）观察到。似乎，鱼类对环境温度的响应，动态改变了许多细胞成分。据认为，环境温度信号被传播到中枢神经系统，然后通过各种马匹法规对基因表达作用。这项研究的目的是披露参与鱼类温度适应的分子机制，并利用在工业应用中获得的结果。将10°C-和30°C型鲤鱼肌球蛋白重链链链同工型的10'上游调节区域的结构完整性鉴定出来，并确定了将MEF2粘合物的鉴定来确定的，并确定了依次的结构。此外，发现来自另一种基本鱼类Medaka的肌球蛋白重链基因的5'上游区域也参与了这种调节以进行温度依赖性基因表达。cARP提高了55 kDa胞质蛋白的表达水平与适应于冷温度的55 kDa胞质蛋白。发现该蛋白是线粒体FOF1-ATPase的亚基之一，并在温度从30°降至10℃时增加了两倍。根据适应温度的温度调节了这种不同的表达。在转录水平上调节了各种鱼类，从Kasumigaura Lake周期左右从Kasumigaura Lake收集，并确定快速骨骼肌肌纤维的ATPase活性。该活性是11月收集的鱼类最高的，并逐渐达到6月和7月的最小值。通过使用对电压敏感的探针和荧光成像，开发了确定线粒体膜电压的分析方法。通过这种方法成功测量了适应鱼环境温度后线粒体膜的电压相关势能的变化。较少的