Tabunoki Hiroko, Dittmer Neal T, Gorman Maureen J, Kanost Michael R
Department of Science of Biological Production, Graduate School of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
Department of Biochemistry and Molecular Biophysics, Kansas State University, 141 Chalmers Hall, Manhattan, KS, 66506-3702, USA.
BMC Res Notes. 2019 Jan 7;12(1):7. doi: 10.1186/s13104-018-4041-y.
Hemolymph plays many important roles in the physiology of an insect throughout its lifetime; however, for small-bodied insects, studies are lacking because of the difficulties encountered while collecting hemolymph. The objective of our study was to develop a method to collect hemolymph plasma from various stages of Tribolium castaneum and to evaluate phenoloxidase activity in the plasma samples. We first designed a procedure for easily and quickly collecting clear hemolymph plasma from T. castaneum.
By using this method, we collected approximately 5 µl plasma from 30 individuals at the larval, pupal or adult stages. And then, we studied the expression of phenoloxidase by performing western blot analysis of the plasma samples and found that phenoloxidase is present in hemolymph in each developmental stage. We also measured phenoloxidase activity in control plasma and plasma treated with Gram-positive bacteria, Micrococcus luteus. Phenoloxidase activity was greater in some of the M. luteus-treated plasma samples compared with control samples. Thus, we developed a method to collect hemolymph plasma that is suitable for studies of phenoloxidase activity.
血淋巴在昆虫的整个生命周期中发挥着许多重要作用；然而，对于小型昆虫来说，由于在收集血淋巴时遇到困难，相关研究较少。我们研究的目的是开发一种从赤拟谷盗不同发育阶段收集血淋巴血浆的方法，并评估血浆样本中的酚氧化酶活性。我们首先设计了一种从赤拟谷盗轻松快速收集清亮血淋巴血浆的程序。
通过使用这种方法，我们从30只幼虫、蛹或成虫阶段的个体中收集了约5微升血浆。然后，我们通过对血浆样本进行蛋白质免疫印迹分析来研究酚氧化酶的表达，发现酚氧化酶存在于每个发育阶段的血淋巴中。我们还测量了对照血浆和用革兰氏阳性菌藤黄微球菌处理的血浆中的酚氧化酶活性。与对照样本相比，一些经藤黄微球菌处理的血浆样本中的酚氧化酶活性更高。因此，我们开发了一种适合于酚氧化酶活性研究的血淋巴血浆收集方法。
