Gene-Environment Interactions in Euryhaline Shore Crabs

蟹属的岸蟹是已知的最具侵袭性的海洋生物之一，很容易在远离其原产地的生态系统中站稳脚跟。这些螃蟹显然是通过交换船舶舱底水而引入的，它们对极端环境具有高度的耐受性，因此不仅能在运输过程中生存，而且在外国目的地也能生存下来。在这些新的环境中，它们的攻击性可以导致生态关系的颠覆性变化。为了更好地了解癌症物种的生物学，拟议的研究将表征欧洲滨蟹基因组中的至少1万个功能基因，这是北美沿海地区常见的非本土物种。一组非冗余的这些基因将以DNA斑点的形式打印在玻璃片上的微阵列中，代表了螃蟹基因组的很大一部分。环境耐受性的最初生理测试将侧重于对极端盐度的适应和相关基因活性的变化。基因产品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。活性基因将在微阵列上作为亮点可见。微阵列结果将通过一种独立的方法进行检查，以开发每个实验处理的基因表达签名。根据Maenas Carcinus的结论将与地中海岸蟹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和渔业研究所的研究人员合作获得。这种对环境刺激响应的基因表达变化的研究可能导致识别非本土岸蟹生理上的薄弱环节，为未来控制种群数量和恢复受这些入侵动物影响的沿海生态系统的生态平衡建立可能的目标。这项拟议的研究是已知的第一次将微阵列分析应用于海洋生物中基因-盐度相互作用的研究。国际和平研究所积极参与培训来自他自己的机构以及美国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，并将包括一名来自克罗地亚的女学生作为这项研究的第一个暑期参与者。这项研究将引起海洋生物学家、比较生理学家以及对基因调控感兴趣的分子生物学家的兴趣。

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.