Changes in the apolipophorin III in Galleria mellonella larvae treated with Pseudomonas aeruginosa exotoxin A

IF 2.3 2区 农林科学 Q1 ENTOMOLOGY
Bartłomiej Iwański, Mariola Andrejko
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Abstract

In the present study, we have demonstrated a correlation in time between changes in the amount of apolipophorin III (apoLp-III) in the fat body and hemocytes of Galleria mellonella larvae challenged with Pseudomonas aeruginosa exotoxin A (exoA). An increase in the amount of apoLp-III was detected 1–8 h after the challenge; then, a temporary decrease was observed after 15 h followed by an increase in the level of apoLp-III, however to a different extent. The profile of apoLp-III forms in the hemolymph, hemocytes, and fat body of the exoA-challenged larvae was analyzed using two-dimensional electrophoresis (IEF/SDS-PAGE) and immunoblotting with anti-apoLp-III antibodies. Two apoLp-III forms differing in isoelectric point values estimated at ∼ 6.5 and ∼ 6.1 in the hemolymph and ∼ 6.5 and ∼ 5.9 in the hemocytes as well as one isoform with pI ∼ 6.5 in the fat body with an additional apoLp-III-derived polypeptide with estimated pI ∼ 6.9 were detected in the control insects. The injection of exoA caused a significant decrease in the abundance of both apoLp-III isoforms in the insect hemolymph. In the hemocytes, a decrease in the amount of the pI ∼ 5.9 isoform was detected, while the major apoLp-III isoform (pI ∼ 6.5) remained unchanged. In addition, appearance of an additional apoLp-III-derived polypeptide with an estimated pI ∼ 5.2 was observed. Interestingly, there were no statistically significant differences in the amount of the main isoform in the fat body between the control and exoA-challenged insects, but the polypeptide with pI ∼ 6.9 disappeared completely. It should be noted that the decrease in the amount of apoLp-III and other proteins was especially noticeable at the time points when exoA was detected in the studied tissues.

Abstract Image

铜绿假单胞菌外毒素A对意大利黑加仑幼虫载脂蛋白III的影响
在本研究中,我们已经证明了用铜绿假单胞菌外毒素a(exoA)攻击的意大利黑加仑幼虫的脂肪体和血细胞中载脂蛋白III(apoLp III)的量在时间上的变化之间的相关性。攻击后1-8小时检测到apoLp III的量增加;然后,在15小时后观察到暂时的下降,随后apoLp III的水平增加,但程度不同。使用二维电泳(IEF/SDS-PAGE)和抗apoLp III抗体的免疫印迹分析了外显子A攻击幼虫的血淋巴、血细胞和脂肪体中apoLpⅢ形式的分布。在对照昆虫中检测到两种等电点值不同的apoLp III形式,血淋巴中估计为~6.5和~6.1,血细胞中估计为-6.5和~5.9,以及脂肪体中一种pI为~6.5的同种型和一种额外的来源于apoLpⅢ的多肽,估计为pI为-6.9。exoA的注射导致昆虫血淋巴中两种apoLp III亚型的丰度显著降低。在血细胞中,检测到pI~5.9亚型的数量减少,而主要的apoLp III亚型(pI~6.5)保持不变。此外,观察到一种额外的apoLp III衍生多肽的出现,估计pI~5.2。有趣的是,对照昆虫和exoA攻击昆虫脂肪体中主要异构体的数量没有统计学上的显著差异,但pI~6.9的多肽完全消失。应该注意的是,apoLp III和其他蛋白质的量的减少在所研究的组织中检测到exoA的时间点特别明显。
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来源期刊
Journal of insect physiology
Journal of insect physiology 生物-昆虫学
CiteScore
4.50
自引率
4.50%
发文量
77
审稿时长
57 days
期刊介绍: All aspects of insect physiology are published in this journal which will also accept papers on the physiology of other arthropods, if the referees consider the work to be of general interest. The coverage includes endocrinology (in relation to moulting, reproduction and metabolism), pheromones, neurobiology (cellular, integrative and developmental), physiological pharmacology, nutrition (food selection, digestion and absorption), homeostasis, excretion, reproduction and behaviour. Papers covering functional genomics and molecular approaches to physiological problems will also be included. Communications on structure and applied entomology can be published if the subject matter has an explicit bearing on the physiology of arthropods. Review articles and novel method papers are also welcomed.
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