Collaborative Research: Role of Endothelial Cell Activation in Hypoxia Tolerance of an Elite Diver, the Weddell Seal

合作研究：内皮细胞激活在精英潜水员威德尔海豹耐缺氧中的作用

• 批准号: 2020664
• 负责人: Jose Pablo Vazquez-Medina
• 金额: $ 50.02万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2020664&HistoricalAwards=false
• 关键词: Collaborative; Research; Role; Endothelial; Cell

This award is funded in whole under the American Rescue Plan Act of 2021 (Public Law 117-2).Part I: Non-technical description: The Weddell seal is an iconic Antarctic species and a superb diver, swimming down to 2,000 feet and staying underwater for up to 45 minutes. However, as for any mammal, the low oxygen concentrations in the blood during diving and the recovery once back at the surface are challenges that need to be overcome making their diving ability something unique that has fascinated scientists for decades. This research project will evaluate the underlying processes in Weddell seal’s physiology that protects this species from the consequences of diving. The work will combine laboratory experiments where cells that line the blood vessels will be exposed to conditions of low oxygen, similar to those that will be measured in diving seals in Antarctica. The investigarors will test a new idea that several short-term dives, performed before a long dive, allows seals to condition themselves. Measurements on the chemical compounds released to the blood during dives, combined with experiments on the genes that regulate them will provide clues on the biochemical pathways that help the seals tolerate these extreme conditions. The project allows for documentation of individual seal dives and provisioning of such information to the broader science community that seeks to study these seals, educating graduate and undergraduate students and a post-doctoral researcher and producing a science-outreach comic book for middle-school students to illustrate the project's science activities, goals and outcomes. Part II: Technical description: The Weddell seal is a champion diver with high natural tolerance for low blood oxygen concentration (hypoxemia) and inadequate blood supply (ischemia). The processes unique to this species protects their tissues from inflammation and oxidative stress observed in other mammalian tissues exposed to such physiological conditions. This project aims to understand the signatures of the processes that protect seals from inflammation and oxidant stress, using molecular, cellular and metabolic tools. Repetitive short dives before long ones are hypothesized to precondition seal tissues and activate the protective processes. The new aspect of this work is the study of endothelial cells, which sense changes in oxygen and blood flow, providing a link between breath-holding and cellular function. The approach is one of laboratory experiments combined with 2-years of field work in an ice camp off McMurdo Station in Antarctica. The study is structured by three main objectives: 1) laboratory experiments with arterial endothelial cells exposed to changes in oxygen and flow to identify molecular pathways responsible for tolerance of hypoxia and ischemia using several physiological, biochemical and genomic tools including CRSPR/Cas9 knochout and knockdown approaches. 2) Metabolomic analyses of blood metabolites produced by seals during long dives. And 3) Metabolomic and genomic determinations of seal physiology during short dives hypothesized to pre-condition tolerance responses. In the field, blood samples will be taken after seals dive in an isolated ice hole and its diving performance recorded. It is expected that the blood will contain metabolites that can be related to molecular pathways identified in lab experiments. Expert collaborators will provide field support, with the ice camp, dive hole for the seals, and telemetry associated with the seals’ dives. The project builds upon previous NSF-funded projects where the seal genome and cellular resources were produced. Undergraduate researchers will be recruited from institutional programs with a track record of attracting underrepresented minorities and a minority-serving institution. To further increase polar literacy training and educational impacts, the field team will include a blog where field experiences are shared and comic book preparation with an artist designed for K-12 students and public outreach.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该奖项根据《2021 年美国救援计划法案》（公法 117-2）提供全额资助。 第一部分：非技术说明：威德尔海豹是南极标志性物种，也是出色的潜水员，可以游到 2,000 英尺深并停留在水中水下长达 45 分钟然而，对于任何哺乳动物来说，潜水期间血液中的低氧浓度以及返回水面后的恢复都是需要克服的挑战，这使得它们的潜水能力成为了问题。这项研究项目将评估威德尔海豹的生理机制，以保护该物种免受潜水的影响。这项工作将结合实验室实验，使血管内壁的细胞暴露在低温条件下。与在南极洲的潜水海豹中测量的氧气类似，研究人员将测试一种新想法，即在长时间潜水之前进行几次短期潜水，以便测量释放到血液中的化学物质。期间潜水，结合对调节它们的基因的实验，将提供有关帮助海豹忍受这些极端条件的生化途径的线索，该项目可以记录个体海豹的潜水情况，并向寻求研究的更广泛的科学界提供此类信息。这些海豹为研究生和本科生以及博士后研究员提供教育，并为中学生制作一本科学推广漫画书，以说明该项目的科学活动、目标和成果第二部分：技术描述：威德尔海豹是冠军。具有高自然耐受性的潜水员该物种独特的过程可以保护其组织免受暴露于此类生理条件的其他哺乳动物组织中观察到的炎症和氧化应激的影响。这项工作的新方面是利用分子、细胞和代谢工具来预处理密封组织并激活保护过程，从而保护海豹免受炎症和氧化应激的影响。该研究是在南极洲麦克默多站附近的一个冰营中进行的实验室实验与两年的实地工作相结合的结果。其结构由三个主要目标组成：1）利用暴露于氧气和流量变化的动脉内皮细胞进行实验室实验，以使用多种生理、生化和基因组工具来识别负责缺氧和缺血耐受的分子途径，包括CRSPR/Cas9 敲除和敲低方法 2) 对海豹在长时间潜水期间产生的血液代谢物进行代谢组学分析，以及 3) 对在现场采集的海豹生理学进行代谢组学和基因组测定，以达到预先条件耐受反应。在海豹潜入一个孤立的冰洞并记录其潜水表现后，专家合作者将提供与实验室实验中分子识别途径相关的代谢物。支持，包括冰营、海豹潜水洞以及与海豹潜水相关的遥测技术。该项目建立在以前由国家科学基金会资助的项目的基础上，这些项目将从机构项目中招募本科生研究人员。吸引代表性不足的少数群体和为少数群体服务的机构的记录 为了进一步提高极地扫盲培训和教育影响，现场团队将包括一个分享现场经验的博客，并与专为 K-12 学生设计的漫画书一起制作。公共宣传。此奖项通过使用基金会的智力价值和更广泛的影响审查标准进行评估，NSF 的法定使命被认为值得支持。