Giannetto Alessia, Maisano Maria, Cappello Tiziana, Oliva Sabrina, Parrino Vincenzo, Natalotto Antonino, De Marco Giuseppe, Barberi Chiara, Romeo Orazio, Mauceri Angela, Fasulo Salvatore
Department of Biological and Environmental Sciences, University of Messina, 98166, Messina, Italy.
Mar Biotechnol (NY). 2015 Dec;17(6):768-81. doi: 10.1007/s10126-015-9655-7. Epub 2015 Aug 16.
Aquatic organisms experience environmental hypoxia as a result of eutrophication and naturally occurring tidal cycles. Mytilus galloprovincialis, being an anoxic/hypoxic-tolerant bivalve, provides an excellent model to investigate the molecular mechanisms regulating oxygen sensing. Across the animal kingdom, inadequacy in oxygen supply is signalled predominantly by hypoxia-inducible factors (HIF) and Hif-prolyl hydroxylases (PHD). In this study, hif-α 5'-end and partial phd mRNA sequences from M. galloprovincialis were obtained. Phylogenetic and molecular characterization of both HIF-α and PHD putative proteins showed shared key features with the respective orthologues from animals strongly suggesting their crucial involvement in the highly conserved oxygen sensing pathway. Both transcripts displayed a tissue-specific distribution with prominent expression in gills. Quantitative gene expression analysis of hif-α and phd mRNAs from gills of M. galloprovincialis demonstrated that both these key sensors are transcriptionally modulated by oxygen availability during the short-time air exposure and subsequent re-oxygenation treatments proving that they are critical players of oxygen-sensing mechanisms in mussels. Remarkably, hif-α gene expression showed a prompt and transient response suggesting the precocious implication of this transcription factor in the early phase of the adaptive response to hypoxia in Mytilus. HIF-α and PHD proteins were modulated in a time-dependent manner with trends comparable to mRNA expression patterns, thus suggesting a central role of their transcriptional regulation in the hypoxia tolerance strategies in marine bivalves. These results provide molecular information about the effects of oxygen deficiency and identify hypoxia-responsive biomarker genes in mussels applicable in ecotoxicological studies of natural marine areas.
由于富营养化和自然发生的潮汐周期,水生生物会经历环境缺氧。作为一种耐缺氧/低氧的双壳贝类,地中海贻贝为研究调节氧感应的分子机制提供了一个极佳的模型。在整个动物界,氧气供应不足主要由缺氧诱导因子(HIF)和Hif-脯氨酰羟化酶(PHD)发出信号。在本研究中,获得了地中海贻贝的hif-α 5'端和部分phd mRNA序列。HIF-α和PHD推定蛋白的系统发育和分子特征显示,它们与来自其他动物的各自直系同源物具有共同的关键特征,这强烈表明它们在高度保守的氧感应途径中起着至关重要的作用。两种转录本均呈现组织特异性分布,在鳃中表达显著。对地中海贻贝鳃中hif-α和phd mRNA的定量基因表达分析表明,在短期空气暴露和随后的复氧处理过程中,这两种关键传感器均受到氧可用性的转录调节,证明它们是贻贝氧感应机制的关键参与者。值得注意的是,hif-α基因表达显示出迅速而短暂的反应,表明该转录因子在贻贝对缺氧的适应性反应早期就有早熟的影响。HIF-α和PHD蛋白以时间依赖性方式受到调节,其趋势与mRNA表达模式相当,因此表明它们的转录调节在海洋双壳贝类的耐缺氧策略中起着核心作用。这些结果提供了有关缺氧影响的分子信息,并鉴定了贻贝中可用于天然海洋区域生态毒理学研究的缺氧反应生物标志物基因。