Correlating cardiac thermal performance limits with transcriptome profiles during thermal acclimation of the intertidal porcelain crab, Petrolisthes cinctipes

潮间带瓷蟹 Petrolisthes cinctipes 热适应过程中心脏热性能限制与转录组图谱的关联

• 批准号: 0444713
• 负责人: Jonathon Stillman
• 金额: --
• 依托单位: University of Hawaii
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0444713&HistoricalAwards=false
• 关键词: Correlating; cardiac; thermal; performance; limits

Correlating cardiac thermal performance limits with transcriptome profiles during thermal acclimation of the intertidal porcelain crab, Petrolisthes cinctipesJonathon H. StillmanUniversity of Hawaii at ManoaUnderstanding the plasticity and limits of physiological responses to changes in temperature is a key component to understanding how global climate change will impact species distribution and abundance patterns. The aim of this project is to combine functional genomics and cardiac physiology to study the physiological responses of intertidal zone porcelain crabs to environmentally relevant temperature changes. Marine intertidal zone organisms experience changes in habitat temperature over both long (e.g., seasonal) and short (e.g., daily) time scales. This research will characterize physiological acclimatization to seasonal changes in habitat temperature and physiological stress responses following acute temperature changes during a low tide period, both of which are important components of the thermal responses of organisms in their natural habitat.Four specific questions will be addressed: 1) What genes are involved in setting cardiac thermal performance limits?, 2) Does thermal acclimation influence responses to acute thermal stress?, 3) What gene expression and thermal physiology changes occur during acclimation to fluctuating temperatures?, and 4) Does response to acute thermal stress vary across seasons? To answer question #1, crabs will be sampled during a one-month acclimation to three temperatures (6C, 14C, and 22C). Sampling will consist of in vivo determination of upper and lower thermal limits of cardiac function (CTmax and CTmin, respectively), and analysis of heart tissue transcriptome profiles using a custom microarray consisting of 13,824 cloned P. cinctipes cDNAs. To answer question #2, crabs acclimated to those three temperatures will be exposed to an acute heat, cold, or control stress and transcriptome profiles during recovery from those stresses will be analyzed. To answer question #3, crabs will be acclimated for one month to a thermal regime that includes a brief daily exposure to extreme heat or cold (-1C and 30C), after which time CTmax, CTmin and transcriptome profiles will be analyzed during four cycles of the acclimation. To answer question #4, crabs will be sampled from their natural habitat across the seasons of one year and CTmax, CTmin, and transcriptome profiles of field and stressed crabs will be measured during each season. Statistical analysis of transcriptome profiles using microarray ANOVA will reveal which genes have significant changes in expression in each of the four experiments. The data generated in this project will be made publicly available on the Stanford Microarray Database.This research will generate novel hypotheses in fundamental areas of animal physiology, namely the molecular bases of thermal limits, eurythermality, and thermal acclimation. The results of this work will directly inform comparative functional genomics studies of temperature adaptation in porcelain crab congeneric species from a range of thermal habitats across a wide range of biogeographic regions. Thus these data will contribute to our understanding of how global temperature change influences species range distributions.Undergraduate students from Hawaiian and Pacific Islander underrepresented minority groups will be recruited to work on this project through the University of Hawaii's Haumana Minority Access to Research Careers (MARC) programs. Benefits to graduate and undergraduate students involved in this research include the development of laboratory and professional skills, and exposure to the "mainland" scientific community at professional meetings and during summer field-work at the Oregon Institute of Marine Biology.

相关的心脏热性能限制与转录组配置文件的热驯化过程中的潮间带瓷蟹，岩蟹cinctipesJonesH。夏威夷大学斯蒂尔曼分校马诺了解温度变化的生理反应的可塑性和限制是了解全球气候变化如何影响物种分布和丰度模式的关键组成部分。 本项目旨在将联合收割机功能基因组学和心脏生理学相结合，研究潮间带瓷蟹对环境相关温度变化的生理反应。 海洋潮间带生物经历了长期的栖息地温度变化（例如，季节性的）和短的（例如，每日）时间尺度。 本研究将探讨生物对栖息地温度季节性变化的生理适应和低潮期温度急剧变化后的生理应激反应，这两者都是生物在自然栖息地热反应的重要组成部分。本研究将解决四个具体问题：1）哪些基因参与了心脏热性能极限的设定？2)热适应是否影响对急性热应激的反应？3)在适应温度变化的过程中，基因表达和热生理发生了什么变化？（4）对急性热应激的反应是否随季节而变化？ 为了回答问题#1，螃蟹将在一个月的适应三种温度（6 ℃，14 ℃和22 ℃）期间进行采样。 采样将包括体内测定心脏功能的热上限和热下限（分别为CTmax和CTmin），以及使用由13，824个克隆的P. cinctipes cDNA组成的定制微阵列分析心脏组织转录组谱。 为了回答问题#2，适应这三种温度的螃蟹将暴露于急性热、冷或对照应激，并将分析从这些应激恢复期间的转录组谱。 为了回答问题#3，螃蟹将适应一个月的热状态，包括每天短暂暴露于极热或极冷（-1 ℃和30 ℃），之后将在四个适应周期内分析CTmax、CTmin和转录组谱。 为了回答问题#4，将在一年的季节内从其自然栖息地对螃蟹进行采样，并在每个季节期间测量田间和应激螃蟹的CTmax、CTmin和转录组谱。 使用微阵列ANOVA的转录组谱的统计分析将揭示哪些基因在四个实验的每一个中具有显著的表达变化。 该项目产生的数据将在斯坦福大学微阵列数据库中公开，这项研究将在动物生理学的基础领域产生新的假设，即热极限、广温性和热适应的分子基础。 这项工作的结果将直接通知比较功能基因组学研究的温度适应瓷蟹同类物种从一系列的热栖息地在广泛的地理区域。因此，这些数据将有助于我们了解全球温度变化如何影响物种的范围distributions.Undergraduate学生从夏威夷和太平洋岛民代表性不足的少数群体将被招募到这个项目通过夏威夷的夏威夷大学的少数民族访问研究职业（MARC）计划。 参与这项研究的研究生和本科生的好处包括实验室和专业技能的发展，并在专业会议和夏季实地考察期间在俄勒冈州海洋生物研究所接触“大陆”科学界。