Heart Flies - effect of microgravity on heart function in Drosophila

Canton S laboratory wildtype lines (Bloomington Stock Center) and a K+ channel mutant seizure (seizurets1, a gift from Dr. Barry Ganetsky, published in Titus et al., 1997), a homolog of human hERG, were used in this study. Flies launched to the International Space Station and the corresponding ground controls were housed in polystyrene vials (75 x24 mm) with 10 ml of standard cornmeal-based fly food containing double the usual amount of 10% Tegosept and cellulose acetate plugs. Approximately 24 hours prior to launch, 15 vials were each loaded with 10 virgin females and 5 male adult flies. Loaded vials were placed in a Plexiglas tray and inserted into a vented fly box (VFB), a modified version of a Nanoracks box. The VFB was placed into a canvas case for loading into the Dragon capsule. Launch of SpaceX CRS-3 occurred at 19:25 UTC on April 18, 2014. Flies were kept in the VFB throughout the 30-day mission and were stored aboard the docked Dragon capsule that shared atmosphere with the ISS. The Dragon capsule unberthed at 13:26 UTC (Co-ordinated Universal Time) on May 18, 2014 and re-entry into the Earth’s atmosphere occurred the same day at 19:05 UTC. Following retrieval of the Dragon capsule off the coast of Mexico, the VFB was offloaded at Long Beach Port, California and transported by car to La Jolla, California in a climate-controlled vehicle. Surviving adult flies born on the ISS were collected from the vials at 23:30 UTC on May 20, 2014. Data collection, which included negative geotaxis assays, heart function studies, tissue collection for RNAseq, and tissue preservation for immunohistochemistry, was completed by 07:30 UTC on May 21, 2014 on all microg spaceflown and corresponding ground control flies. The VFB was oriented such that the g‐load was directed down into the food supply located at the bottom of the vials. This orientation was maintained throughout the entirety of the mission, from point of transfer at the lab at NASA Kennedy Space Center through stowage on the Dragon capsule, and again from Dragon un-berthing to opening of the VFB in the lab at SBP Medical Discovery Institute in La Jolla, California. Ground control flies were prepared and reared under identical conditions in ground-based incubators with temperature and humidity controlled to levels recorded on the ISS.

本研究使用了Canton S实验室野生型果蝇品系（源自布卢明顿果蝇种质中心Bloomington Stock Center），以及钾离子通道突变体seizurets1——该突变体由Barry Ganetsky博士惠赠，相关成果发表于Titus等人1997年的研究，且为人类hERG基因的同源物。送往国际空间站（International Space Station, ISS）的果蝇及对应的地面对照组果蝇均饲养于聚苯乙烯培养瓶（75×24 mm）中，瓶内添加10 mL标准玉米粉果蝇培养基，该培养基中10% Tegosept的添加量为常规用量的两倍，并配有醋酸纤维素棉塞。发射前约24小时，每个培养瓶中接种10只未交配雌性果蝇与5只成年雄性果蝇。接种后的培养瓶被放置于有机玻璃托盘内，再装入透气果蝇培养箱（vented fly box, VFB）——该装置为Nanoracks箱的改良版本。随后，VFB被装入帆布收纳箱，以便转运至“龙”货运飞船（Dragon capsule）舱内。本研究所用果蝇随SpaceX CRS-3任务于2014年4月18日世界协调时（Coordinated Universal Time, UTC）19:25发射升空。整个30天的太空任务期间，果蝇均留存于VFB中，并存放于与国际空间站大气相通的对接状态龙飞船舱内。龙飞船于2014年5月18日世界协调时13:26与空间站分离，并于当日世界协调时19:05再入地球大气层。龙飞船于墨西哥近海回收后，VFB在加利福尼亚州长滩港卸载，并通过温控车辆转运至加利福尼亚州拉霍亚市。2014年5月20日世界协调时23:30，研究人员从培养瓶中收集了在国际空间站内孵化存活的成年果蝇。所有太空飞行（微重力环境）组及对应地面对照组果蝇的相关数据采集工作于2014年5月21日世界协调时07:30前完成，采集内容包括负趋地性运动实验、心脏功能检测、用于RNA测序（RNAseq）的组织样本采集，以及用于免疫组织化学染色的组织固定保存。VFB的安装方向需保证重力加速度载荷向下指向培养瓶底部的培养基。该方向在整个任务周期内均保持不变：从美国国家航空航天局（National Aeronautics and Space Administration, NASA）肯尼迪航天中心实验室的转运环节，到龙飞船舱内的存放阶段，再到龙飞船分离后于加利福尼亚州拉霍亚市SBP医学发现研究所实验室打开VFB的全过程，均严格维持该朝向。地面对照组果蝇的制备与饲养条件与太空组完全一致，饲养环境的温湿度参数均设置为国际空间站在轨记录的数值。

搜集汇总

背景与挑战

背景概述

该数据集研究了微重力对果蝇心脏功能的影响，使用Canton S野生型和seizurets1突变体果蝇，在国际空间站进行30天微重力暴露实验，并设置了地面对照组。数据主要通过RNA测序技术生成，包括13个样本的转录组数据，旨在分析微重力条件下心脏相关基因表达的变化，为空间生物学和心脏功能研究提供基础。