Department of Comparative Physiology, Uppsala University, Norbyvägen 18 A, SE752 36 Uppsala, Sweden.
Department of Comparative Physiology, Science for Life Laboratory, Uppsala University, Norbyvägen 18 A, SE752 36 Uppsala, Sweden.
Int J Mol Sci. 2021 Jun 16;22(12):6464. doi: 10.3390/ijms22126464.
Early changes in hemocyte proteins in freshwater crayfish , in response to an injection with the fungal pattern recognition protein β-1,3-glucan (laminarin) were investigated, as well as changes after saline (vehicle) injection and in naïve animals. Injection of saline resulted in rapid recruitment of granular hemocytes from surrounding tissues, whereas laminarin injection on the other hand induced an initial dramatic drop of hemocytes. At six hours after injection, the hemocyte populations therefore were of different composition. The results show that mature granular hemocytes increase in number after saline injection as indicated by the high abundance of proteins present in granular cell vesicles, such as a vitelline membrane outer layer protein 1 homolog, mannose-binding lectin, masquerade, crustin 1 and serine protease homolog 1. After injection with the β-1,3-glucan, only three proteins were enhanced in expression, in comparison with saline-injected animals and uninjected controls. All of them may be associated with immune responses, such as a new and previously undescribed Kazal proteinase inhibitor. One interesting observation was that the clotting protein was increased dramatically in most of the animals injected with laminarin. The number of significantly affected proteins was very few after a laminarin injection when compared to uninjected and saline-injected crayfish. This finding may demonstrate some problematic issues with gene and protein expression studies from other crustaceans receiving injections with pathogens or pattern recognition proteins. If no uninjected controls are included and no information about hemocyte count (total or differential) is given, expressions data for proteins or mRNAs are very difficult to properly interpret.
本研究调查了淡水小龙虾血细胞蛋白对真菌模式识别蛋白β-1,3-葡聚糖（昆布多糖）注射的早期变化，以及盐水（载体）注射和未处理动物的变化。盐水注射导致颗粒血细胞从周围组织迅速募集，而另一方面，昆布多糖注射则导致血细胞数量最初急剧下降。因此，注射后 6 小时，血细胞群体的组成不同。结果表明，成熟的颗粒血细胞数量增加，如卵黄膜外层蛋白 1 同源物、甘露糖结合凝集素、伪装、壳聚糖 1 和丝氨酸蛋白酶同源物 1 等颗粒细胞囊泡中存在的大量蛋白质所表明的那样。与盐水注射动物和未注射对照相比，β-1,3-葡聚糖注射后仅三种蛋白质的表达增强。所有这些都可能与免疫反应有关，例如一种新的、以前未描述的 Kazal 蛋白酶抑制剂。一个有趣的观察结果是，大多数注射昆布多糖的动物中凝血蛋白显著增加。与未注射和盐水注射的小龙虾相比，注射昆布多糖后受影响的蛋白质数量非常少。这一发现可能表明，在接受病原体或模式识别蛋白注射的其他甲壳类动物中，进行基因和蛋白质表达研究存在一些问题。如果不包括未注射对照，并且未提供关于血细胞计数（总计数或差异计数）的信息，则蛋白质或 mRNA 的表达数据很难正确解释。
