Gene-Environment Interactions in Euryhaline Shore Crabs

巨蟹属的海岸是已知的最具入侵性的海洋生物之一，它们很容易在远离原生地的生态系统中建立自己。 这些螃蟹显然是通过船舶舱底水的交换引入的，它们对极端环境具有高度的耐受性，因此不仅可以在运输过程中生存，而且可以在外国目的地生存。 在这些新的环境中，它们的侵略性可能导致生态关系的破坏性变化。 为了更好地了解Carcinus物种的生物学，拟议的研究将描述欧洲海岸蟹Carcinus maenas基因组中至少10，000个功能基因的特征，Carcinus maenas是北美沿海地区常见的非本土物种。 这些基因的非冗余集将作为DNA点打印在载玻片上的微阵列中，代表螃蟹基因组的重要部分。 对环境耐受性的初步生理测试将侧重于对极端盐度的适应和基因活动的相关变化。 基因产物将从螃蟹组织中提取，用荧光染料标记，然后添加到微阵列中。 活跃的基因将在微阵列上作为亮点可见。 将使用独立的方法检查微阵列结果，以开发每个实验处理的基因表达特征。 基于Carcinus maenas的结论将与地中海海岸Carcinus aestuarii进行比较，后者的组织样本将通过与克罗地亚杜布罗夫尼克海洋学和渔业研究所的研究人员合作获得。 这项研究的基因表达变化，以响应环境刺激可能会导致识别非土著海岸蟹的生理学的薄弱环节，建立可能的目标，为未来的人口控制和恢复生态平衡的沿海生态系统受这些入侵动物。 这项研究代表了微阵列分析在海洋生物基因-盐度相互作用研究中的第一个已知应用。PI积极参与培训本机构以及美国内外其他机构的本科生和研究生，并制定了分子技术培训的核心课程，并将包括来自克罗地亚的一名女学生，她将成为该研究的第一名夏季参与者。这项研究将对海洋生物学家，比较生理学家以及对基因调控感兴趣的分子生物学家感兴趣。

Shore crabs in the genus Carcinus are among the most invasive marine organisms known, easily establishing themselves in ecosystems far from their native origins. Apparently introduced through exchanges of ships bilge water, these crabs are highly tolerant of environmental extremes and thus can survive not only conditions during transport but also at their foreign destination. In these new environments, their aggressive nature can lead to disruptive changes in ecological relationships. To better understand the biology of Carcinus species, the proposed research will characterize at least 10,000 functional genes in the genome of the European shore crab Carcinus maenas, a common non-indigenous species in coastal regions of North America. A non-redundant set of these genes will be printed as DNA spots in microarrays on glass slides, representing a significant fraction of the crab genome. The initial physiological tests of environmental tolerance will focus on adaptation to salinity extremes and associated changes in gene activity. Gene products will be extracted from crab tissues, labeled with a fluorescent dye, and then added to the microarrays. Active genes will be visible as bright spots on the microarrays. Microarray results will be checked with an independent method to develop a gene expression signature for each experimental treatment. Conclusions based on Carcinus maenas will be compared with the Mediterranean shore crab Carcinus aestuarii, whose tissue samples will be obtained through collaboration with investigators at the Institute of Oceanography and Fisheries in Dubrovnik, Croatia. This study of gene expression changes in response to environmental stimuli may lead to the identification of weak links in the physiology of non-indigenous shore crabs, establishing possible targets for future population control and restoration of ecological balance in coastal ecosystems affected by these invasive animals. The proposed research represents the first known application of microarray analysis to a study of gene-salinity interactions in marine organisms. The PI is actively involved in training undergraduate and graduate students from his own institution as well as other institutions within and outside of the USA, and has developed a core curriculum for molecular techniques training and will include a female student from Croatia be the first summer participant in the research. This research will be of interest to marine biologists, comparative physiologists as well as molecular biologists interested in gene regulation.