Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, 113-8657, Japan.
J Comp Physiol B. 2010 Jan;180(1):105-16. doi: 10.1007/s00360-009-0389-6. Epub 2009 Jul 19.
A monogonont rotifer Brachionus plicatilis has been widely used as a model organism for physiological, ecological studies and for ecotoxicology. Because of the availability of parthenogenetic mode of reproduction as well as its versatility to be used as live food in aquaculture, the population dynamic studies using the rotifer have become more important and acquired the priority over those using other species. Although many studies have been conducted to identify environmental factors that influence rotifer populations, the molecular mechanisms involved still remain to be elucidated. In this study, gene(s) differentially expressed by calorie restriction in the rotifer was analyzed, where a calorie-restricted group was fed 3 h day(-1) and a well-fed group fed ad libitum. A subtracted cDNA library from the calorie-restricted rotifer was constructed using suppression subtractive hybridization (SSH). One hundred sixty-three expressed sequence tags (ESTs) were identified, which included 109 putative genes with a high identity to known genes in the publicly available database as well as 54 unknown ESTs. After assembling, a total of 38 different genes were obtained among 109 ESTs. Further validation of expression by semi-quantitative reverse transcription-PCR showed that 29 out of the 38 genes obtained by SSH were up regulated by calorie restriction.
褶皱臂尾轮虫(Brachionus plicatilis)是一种单巢轮虫,已被广泛用作生理、生态研究和生态毒理学的模式生物。由于它具有孤雌生殖的繁殖方式,以及作为水产养殖活饵料的多功能性,使用轮虫进行种群动态研究变得更加重要,并优先于使用其他物种进行的研究。尽管已经进行了许多研究来确定影响轮虫种群的环境因素,但涉及的分子机制仍有待阐明。在这项研究中,分析了轮虫中因热量限制而差异表达的基因,其中热量限制组每天喂食 3 小时,而饱食组则自由喂食。使用抑制性消减杂交(SSH)构建了来自热量限制轮虫的 cDNA 文库。鉴定了 163 个表达序列标签(EST),其中包括 109 个与公共数据库中已知基因具有高度同源性的假定基因,以及 54 个未知 EST。组装后,在 109 个 EST 中总共获得了 38 个不同的基因。通过半定量逆转录-PCR 进一步验证表达,发现 SSH 获得的 38 个基因中有 29 个受到热量限制的上调。