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Hemocyanin and hexamerins expression in response to hypoxia in stoneflies (Plecoptera, Insecta)
Archives of Insect Biochemistry and Physiology ( IF 2.2 ) Pub Date : 2020-09-26 , DOI: 10.1002/arch.21743
Maribet Gamboa 1
Affiliation  

Many freshwater ecosystems worldwide undergo hypoxia events that can trigger physiological, behavioral, and molecular responses in many organisms. Among such molecular responses, the regulation of the hemocyanin (Hc) protein expression which plays a major role in oxygen transportation within aquatic insects remains poorly understood. The stoneflies (Plecoptera) are aquatic insects that possess a functional Hc in the hemolymph similar to crustacean that co‐occurs with a nonfunctional Hc protein, hexamerins (Hx). However, the role of both proteins during hypoxia remains undetermined. Here, we evaluated the effect of hypoxia on the expression of Hc and Hx proteins via a comparison between hypoxia and normoxia amino acid sequence variation and protein expression pattern within 23 stonefly species. We induced short‐term hypoxia in wild‐caught stoneflies species, sequenced the target region of Hc and Hx by complementary DNA synthesis, characterized the protein biochemistry using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, ultrafiltration, and polarographic fluorometric method, and amplified the genome region of the hypoxia‐inducible factor (HIF) transcriptional response element that regulated Hc using genome walking library approach. We found a lack of Hc expression in all examined species during hypoxia conditions, despite recognition of the HIF gene region as a possible regulatory factor of Hc, suggesting that compensatory responses as metabolic changes or behavioral tracheal movements to enhance respiratory efficiency could be possible mechanics to compensate for hypoxia. A short Hc‐like novel isoform was detected instead in these 23 species, possibly due to either protein degradation or alternative splicing mechanisms, suggesting that the protein could be performing a different function other than oxygen transportation. Hx during hypoxia was expressed and exhibited species‐level amino acid changes, highlighting a possible role during hypoxia. Our results demonstrate that hypoxia could enable a similar potential adaptive response of multiple species regarding specific physiological requirements, thereby shedding light on community behavior in stress environments that may help us to improve conservation practices and biomonitoring.

中文翻译:

血红蛋白和六聚体蛋白表达对低氧的响应(Plecoptera,Insecta)

全球许多淡水生态系统都发生缺氧事件,这些事件可以触发许多生物体的生理,行为和分子反应。在这样的分子反应中,对血蓝蛋白(Hc)蛋白表达的调节在水生昆虫内的氧气运输中起主要作用仍然知之甚少。石蝇(Plecoptera)是一种水生昆虫,其在血淋巴中具有功能性Hc,类似于甲壳类动物,而甲壳类动物与非功能性Hc蛋白六聚体(Hx)同时出现。但是,两种蛋白在缺氧过程中的作用仍未确定。在这里,我们通过比较23个石蝇种类中的缺氧和正常氧氨基酸序列变异和蛋白质表达模式,评估了缺氧对Hc和Hx蛋白表达的影响。我们在野生捕捞的蝇类中诱导了短期缺氧,通过互补DNA合成对Hc和Hx的靶区域进行了测序,使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,超滤和极谱荧光法对蛋白质的生物化学进行了表征,并扩增了基因组使用基因组步行文库方法调节Hc的低氧诱导因子(HIF)转录反应元件的区域。我们发现在缺氧条件下所有受检物种中均缺乏Hc表达,尽管已将HIF基因区识别为可能的Hc调节因子,这表明作为代谢变化或行为性气管运动以增强呼吸效率的代偿反应可能是补偿缺氧。相反,在这23个物种中检测到了一种短的Hc样新型同工型,这可能是由于蛋白质降解或其他剪接机制所致,这表明该蛋白质可能具有除氧气转运以外的其他功能。缺氧期间的Hx被表达并表现出物种水平的氨基酸变化,从而突出了缺氧期间的可能作用。我们的结果表明,低氧可以使多种物种在特定的生理需求方面具有类似的潜在适应性反应,从而揭示应激环境中的群落行为,这可能有助于我们改善保护措施和生物监测。提示该蛋白质可能除了氧气运输外还具有其他功能。缺氧期间的Hx被表达并表现出物种水平的氨基酸变化,从而突出了缺氧期间的可能作用。我们的结果表明,低氧可以使多种物种在特定的生理需求方面具有类似的潜在适应性反应,从而揭示应激环境中的群落行为,这可能有助于我们改善保护措施和生物监测。提示该蛋白质可能除了氧气运输外还具有其他功能。缺氧期间的Hx被表达并表现出物种水平的氨基酸变化,从而突出了缺氧期间的可能作用。我们的研究结果表明,低氧可以使多种物种在特定的生理需求方面具有类似的潜在适应性反应,从而揭示应激环境下的群落行为,这可能有助于我们改善保护措施和生物监测。
更新日期:2020-10-12
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