Larade Kevin, Storey Kenneth B
Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, Ontario, Canada, K1S 5B6.
J Exp Biol. 2004 Mar;207(Pt 8):1353-60. doi: 10.1242/jeb.00872.
Differential screening of a Littorina littorea (the common periwinkle) cDNA library identified ferritin heavy chain as an anoxia-induced gene in hepatopancreas. Northern blots showed that ferritin heavy chain transcript levels were elevated twofold during anoxia exposure, although nuclear run-off assays demonstrated that ferritin heavy chain mRNAs were not transcriptionally upregulated during anoxia. Polysome analysis indicated that existing ferritin transcripts were actively translated during the anoxic period. This result was confirmed via western blotting, which demonstrated a twofold increase in ferritin heavy chain protein levels during anoxia, with a subsequent decrease to control levels during normoxic recovery. Organ culture experiments using hepatopancreas slices demonstrated a >50% increase in ferritin heavy chain transcript levels in vitro under conditions of anoxia and freezing, as well as after incubation with the second messenger cGMP. Taken together, these results suggest that ferritin heavy chain is actively regulated during anoxia exposure in the marine snail, L. littorea.
对滨螺(Littorina littorea,即常见的滨螺)cDNA文库进行差异筛选,确定铁蛋白重链是肝胰腺中的一种缺氧诱导基因。Northern印迹显示,在缺氧暴露期间,铁蛋白重链转录本水平升高了两倍,尽管核转录分析表明,在缺氧期间铁蛋白重链mRNA并未在转录水平上上调。多核糖体分析表明,现有的铁蛋白转录本在缺氧期间被积极翻译。通过蛋白质免疫印迹证实了这一结果,该分析表明在缺氧期间铁蛋白重链蛋白水平增加了两倍,随后在常氧恢复期间降至对照水平。使用肝胰腺切片进行的器官培养实验表明,在缺氧、冷冻条件下以及与第二信使cGMP孵育后,体外铁蛋白重链转录本水平增加了50%以上。综上所述,这些结果表明,在海洋蜗牛滨螺缺氧暴露期间,铁蛋白重链受到积极调控。