Live imaging of Aiptasia larvae, a model system for coral and anemone bleaching, using a simple microfluidic device
Live imaging of Aiptasia larvae, a model system for coral and anemone bleaching, using a simple microfluidic device
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DOI：
10.1038/s41598-019-45167-2
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发表时间：
2019-06-25
影响因子：
4.6
通讯作者：
Fordyce, Polly M.
中科院分区：
文献类型：
--
作者：
Van Treuren, Will;Brower, Kara K.;Fordyce, Polly M.
珊瑚礁及其相关的各种生态系统具有巨大的生态重要性。近年来，珊瑚健康受到人类活动和气候变化带来的环境压力的严重影响，威胁到几个主要珊瑚礁生态系统的灭绝。珊瑚礁的损害是由一个过程介导的所谓的“珊瑚漂白”，珊瑚，海葵，和其他刺胞动物失去了光合藻类共生体（家庭共生藻）后，压力诱导，导致大幅减少主机的能量收获，并最终，珊瑚死亡。这种重要的刺胞藻共生关系消失的机制仍然知之甚少。海葵的幼虫，Exaiptasia pallida（通常被称为“Aiptasia”）是一个有吸引力的模式生物来研究这个过程，但他们是大的（类似于100毫米的长度，类似于75毫米的直径），可变形的，和高度能动的，复杂的长期成像和限制研究这种关键的内共生关系在活的生物体。在这里，我们报告'Traptasia'，一个简单的微流体装置，多个陷阱，旨在隔离和成像个人，活幼虫Aiptasia和他们的藻类共生体在延长的时间课程。使用通过流体流动模拟和聚合物珠加载测试参数化的陷阱设计，我们捕获了含有藻类共生体的Aiptasia幼虫，并证明了稳定的成像>10小时。我们可视化Aiptasia幼虫内的藻类，并观察环境压力下的藻类驱逐。据我们所知，该设备是第一个能够对刺胞动物幼虫及其藻类共生体进行延时，高通量实时成像的设备，并且在进一步的实施中，可以为不同环境压力下刺胞动物漂白的细胞机制提供重要的见解。“Traptasia”装置使用简单，需要最少的外部设备，无需专门的操作培训，并且可以很容易地使用本文提供的陷阱优化数据来研究各种大型能动生物。
Coral reefs, and their associated diverse ecosystems, are of enormous ecological importance. In recent years, coral health has been severely impacted by environmental stressors brought on by human activity and climate change, threatening the extinction of several major reef ecosystems. Reef damage is mediated by a process called 'coral bleaching' where corals, sea anemones, and other cnidarians lose their photosynthetic algal symbionts (family Symbiodiniaceae) upon stress induction, resulting in drastically decreased host energy harvest and, ultimately, coral death. The mechanism by which this critical cnidarian-algal symbiosis is lost remains poorly understood. The larvae of the sea anemone, Exaiptasia pallida (commonly referred to as 'Aiptasia') are an attractive model organism to study this process, but they are large (similar to 100 mm in length, similar to 75 mm in diameter), deformable, and highly motile, complicating long-term imaging and limiting study of this critical endosymbiotic relationship in live organisms. Here, we report 'Traptasia', a simple microfluidic device with multiple traps designed to isolate and image individual, live larvae of Aiptasia and their algal symbionts over extended time courses. Using a trap design parameterized via fluid flow simulations and polymer bead loading tests, we trapped Aiptasia larvae containing algal symbionts and demonstrated stable imaging for >10 hours. We visualized algae within Aiptasia larvae and observed algal expulsion under an environmental stressor. To our knowledge, this device is the first to enable time-lapsed, high-throughput live imaging of cnidarian larvae and their algal symbionts and, in further implementation, could provide important insights into the cellular mechanisms of cnidarian bleaching under different environmental stressors. The 'Traptasia' device is simple to use, requires minimal external equipment and no specialized training to operate, and can easily be adapted using the trap optimization data presented here to study a variety of large, motile organisms.