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利用微阵列技术了解野生宽吻海豚(瓶鼻海豚)暴露于环境污染物中的影响。

Microarray applications to understand the impact of exposure to environmental contaminants in wild dolphins (Tursiops truncatus).

作者信息

Mancia Annalaura, Abelli Luigi, Kucklick John R, Rowles Teresa K, Wells Randall S, Balmer Brian C, Hohn Aleta A, Baatz John E, Ryan James C

机构信息

Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy; Marine Biomedicine and Environmental Science Center, Medical University of South Carolina, Hollings Marine Laboratory, Charleston, SC 29412, USA.

Department of Life Sciences and Biotechnology, University of Ferrara, 44121 Ferrara, Italy.

出版信息

Mar Genomics. 2015 Feb;19:47-57. doi: 10.1016/j.margen.2014.11.002. Epub 2014 Nov 17.

Abstract

It is increasingly common to monitor the marine environment and establish geographic trends of environmental contamination by measuring contaminant levels in animals from higher trophic levels. The health of an ecosystem is largely reflected in the health of its inhabitants. As an apex predator, the common bottlenose dolphin (Tursiops truncatus) can reflect the health of near shore marine ecosystems, and reflect coastal threats that pose risk to human health, such as legacy contaminants or marine toxins, e.g. polychlorinated biphenyls (PCBs) and brevetoxins. Major advances in the understanding of dolphin biology and the unique adaptations of these animals in response to the marine environment are being made as a result of the development of cell-lines for use in in vitro experiments, the production of monoclonal antibodies to recognize dolphin proteins, the development of dolphin DNA microarrays to measure global gene expression and the sequencing of the dolphin genome. These advances may play a central role in understanding the complex and specialized biology of the dolphin with regard to how this species responds to an array of environmental insults. This work presents the creation, characterization and application of a new molecular tool to better understand the complex and unique biology of the common bottlenose dolphin and its response to environmental stress and infection. A dolphin oligo microarray representing 24,418 unigene sequences was developed and used to analyze blood samples collected from 69 dolphins during capture-release health assessments at five geographic locations (Beaufort, NC, Sarasota Bay, FL, Saint Joseph Bay, FL, Sapelo Island, GA and Brunswick, GA). The microarray was validated and tested for its ability to: 1) distinguish male from female dolphins; 2) differentiate dolphins inhabiting different geographic locations (Atlantic coasts vs the Gulf of Mexico); and 3) study in detail dolphins resident in one site, the Georgia coast, known to be heavily contaminated by Aroclor 1268, an uncommon polychlorinated (PCB) mixture. The microarray was able to distinguish dolphins by sex, geographic location, and corroborate previously published health irregularities for the Georgia dolphins. Genes involved in xenobiotic metabolism, development/differentiation and oncogenic pathways were found to be differentially expressed in GA dolphins. The report bridges the advancements in dolphin genome sequencing to the first step towards providing a cost-effective means to screen for indicators of chemical toxin exposure as well as disease status in top level predators.

摘要

通过测量处于较高营养级的动物体内的污染物水平来监测海洋环境并确定环境污染的地理趋势变得越来越普遍。生态系统的健康状况很大程度上反映在其居民的健康状况上。作为顶级捕食者,宽吻海豚(Tursiops truncatus)可以反映近岸海洋生态系统的健康状况,并反映对人类健康构成风险的沿海威胁,如遗留污染物或海洋毒素,例如多氯联苯(PCBs)和短裸甲藻毒素。由于用于体外实验的细胞系的开发、用于识别海豚蛋白质的单克隆抗体的生产、用于测量全球基因表达的海豚DNA微阵列的开发以及海豚基因组的测序,在理解海豚生物学以及这些动物对海洋环境的独特适应性方面取得了重大进展。这些进展可能在理解海豚复杂而特殊的生物学方面发挥核心作用,即该物种如何应对一系列环境侵害。这项工作展示了一种新分子工具的创建、表征和应用,以更好地理解宽吻海豚复杂而独特的生物学及其对环境压力和感染的反应。开发了一种代表24,418个单基因序列的海豚寡核苷酸微阵列,并用于分析在五个地理位置(北卡罗来纳州博福特、佛罗里达州萨拉索塔湾、佛罗里达州圣约瑟夫湾、佐治亚州萨佩洛岛和佐治亚州不伦瑞克)进行捕获-释放健康评估期间从69只海豚采集的血液样本。对该微阵列进行了验证和测试,以检验其能否:1)区分雄性和雌性海豚;2)区分居住在不同地理位置的海豚(大西洋海岸与墨西哥湾);3)详细研究居住在一个地点(佐治亚海岸)的海豚,该地点已知受到一种不常见的多氯联苯混合物Aroclor 1268的严重污染。该微阵列能够按性别、地理位置区分海豚,并证实了先前发表的关于佐治亚海豚健康异常的情况。发现参与异生物质代谢、发育/分化和致癌途径的基因在佐治亚海豚中差异表达。该报告将海豚基因组测序的进展与朝着提供一种经济有效的方法来筛选顶级捕食者中化学毒素暴露指标以及疾病状态的第一步联系起来。

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