McElroy A E, Hice L A, Frisk M G, Purcell S L, Phillips N C, Fast M D
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY, USA.
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY, USA.
Comp Biochem Physiol Part D Genomics Proteomics. 2015 Jun;14:53-65. doi: 10.1016/j.cbd.2015.01.006. Epub 2015 Feb 4.
Inshore winter flounder (Pseudoplueronectes americanus) populations in NY, USA have reached record low numbers in recent years, and recruitment into the fishery appears to be limited by survival of post-settlement juvenile fish. In order to identify cellular pathways associated with site-specific variation in condition and mortality, we examined differential mRNA expression in juvenile winter flounder collected from six different bays across a gradient in human population density and sewage inputs. Illumina sequencing of pooled samples of flounder from contrasting degraded sites and less impacted sites was used to guide our choice of targets for qPCR analysis. 253 transcripts of >100bp were differentially expressed, with 60% showing strong homology to mostly teleost sequences within the NCBI database. Based on these data, transcripts representing nine genes of interest associated with contaminant exposure, immune response and glucose and glycogen metabolism were examined by qPCR in individual flounder from each site. Statistically significant site-specific differences were observed in expression of all but one gene, although patterns in expression were complex with only one (vitellogenin), demonstrating a west to east gradient consistent with known loadings of municipal sewage effluent. Principal components analysis (PCA) identified relationships among the genes evaluated. Our data indicate that juvenile winter flounder are responding to estrogenic chemicals in more urbanized coastal bays, and suggests potential mechanistic links between immune response, contaminant exposure and energy metabolism.
美国纽约近岸冬季比目鱼(美洲拟庸鲽)种群数量近年来已降至历史最低水平,渔业补充量似乎受到定居后幼鱼存活率的限制。为了确定与特定地点条件和死亡率差异相关的细胞途径,我们检测了从六个人口密度和污水排放梯度不同的海湾采集的幼年冬季比目鱼的mRNA差异表达。对比退化地点和受影响较小地点的比目鱼混合样本的Illumina测序,用于指导我们选择qPCR分析的目标。253条长度大于100bp的转录本差异表达,其中60%与NCBI数据库中大多数硬骨鱼序列具有高度同源性。基于这些数据,通过qPCR检测了来自每个地点的个体比目鱼中代表与污染物暴露、免疫反应以及葡萄糖和糖原代谢相关的9个感兴趣基因的转录本。除一个基因外,所有基因的表达均观察到具有统计学意义的特定地点差异,尽管表达模式复杂,只有一个基因(卵黄蛋白原)呈现出从西到东的梯度,与已知的城市污水排放负荷一致。主成分分析(PCA)确定了所评估基因之间的关系。我们的数据表明,幼年冬季比目鱼正在对城市化程度更高的沿海海湾中的雌激素类化学物质做出反应,并暗示了免疫反应、污染物暴露和能量代谢之间潜在的机制联系。