Testing the Stability of Plasma Protein and Whole Blood RNA in Archived Blood of Loggerhead Sea Turtles, Caretta caretta
Testing the Stability of Plasma Protein and Whole Blood RNA in Archived Blood of Loggerhead Sea Turtles, Caretta caretta
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测试红海龟存档血液中血浆蛋白和全血 RNA 的稳定性
DOI：
10.1089/bio.2020.0026
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发表时间：
2020
影响因子：
1.6
通讯作者：
Lynch, Jennifer M.
中科院分区：
文献类型：
--
作者：
Townsend, Kelly;Ness, Jennifer;Hoguet, Jennifer;Stacy, Nicole I.;Komoroske, Lisa M.;Lynch, Jennifer M.
样品储存条件会影响生物测量的准确性和再现性。在最低可用温度下快速储存样品被认为是理想的选择，但在偏远和后勤困难的现场条件下采样时并不总是可行，就像海龟的情况一样。本研究的目的是检查红海龟样本中血浆蛋白的稳定性和全血 RNA 的质量，这些样本是历时 18 年精心收集的样本的一部分。这些生物变量通常用于评估海龟的健康状况；因此，有必要在储存过程中保持这些组件的完整性。对 2018 年（n=≥3）、2008 年（n=≥3）和 2001 年（n=≥3）采集的海龟个体肝素化血浆进行蛋白电泳。血浆还来自 2018 年采集的四只海龟样本，并经过不同的储存温度。全血收集在含有肝素钠的采血管或含有 RNA 防腐剂的 PAXgene 管中。这些都经过了不同的存储处理，这些处理可能在后勤困难的现场采样期间发生。经过各种处理后，血浆蛋白在不同收集年份之间表现出微小差异，并且即使在 38°C 下暴露三个小时，也没有观察到储存处理之间的差异。使用 RNA 完整性数 (RIN) 评估全血的 RNA 质量。冷冻的肝素钠管和长时间解冻后的 PAXgene 管的 RIN 较差。从从未冷冻的肝素钠管和冷冻延迟长达 11 天的 PAXgene 管中获得高质量 RNA。总的来说，这些结果表明血浆蛋白随着时间的推移和暴露于不良的储存条件时保持稳定，而海龟血液在冷冻后和保存不当时会迅速降解。在规划采样方案和物流以实现最佳的长期样品保存时，考虑这些方面非常重要。
Sample storage conditions can affect accuracy and reproducibility of biological measurements. Storing samples rapidly at the lowest available temperatures is considered ideal but is not always feasible when sampling in remote and logistically challenging field conditions, as is often the case with sea turtles. The objective of this study was to examine the stability of plasma proteins and quality of whole blood RNA from loggerhead sea turtle samples collected as part of an eighteen-year-long curated specimen collection. These biological variables are often used to assess sea turtle health; therefore, it is necessary to maintain the integrity of these components during storage. Protein electrophoresis was conducted on heparinized plasma from individual turtles collected in 2018 (n= 3), 2008 (n= 3), and 2001 (n= 3). Plasma was also pooled from four turtles sampled in 2018 and subjected to various storage temperatures. Whole blood was collected in blood collection tubes containing sodium heparin or PAXgene tubes with an RNA preservative. These were subjected to different storage treatments that can possibly occur during logistically difficult field sampling. Following various treatments, plasma proteins showed minor differences across collection years and no differences among storage treatments were observed, even when exposed to 38°C for three hours. RNA quality was assessed from whole blood using an RNA integrity number (RIN). RINs were poor from sodium heparin tubes that were frozen and from PAXgene tubes after an extended thaw. High-quality RNA was obtained from sodium heparin tubes that were never frozen and from PAXgene tubes with freezing delayed by up to 11 days. Overall, these results indicate that plasma proteins remain stable over time and when exposed to undesirable storage conditions, and RNA degrades rapidly in sea turtle blood after freezing and when not properly preserved. These aspects are important to consider when planning sampling protocols and logistics for optimal long-term sample preservation.