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犬弓首蛔虫、牛弓首蛔虫和猪蛔虫中精氨酸激酶(AK)的免疫定位。

Immunolocalization of arginine kinase (AK) in Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides.

机构信息

Department of Veterinary Pathobiology, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, Peradeniya, Sri Lanka.

出版信息

Parasitol Res. 2012 Aug;111(2):663-71. doi: 10.1007/s00436-012-2884-z. Epub 2012 Mar 8.

Abstract

Arginine kinase (AK) is a member of the phosphagen kinase family. AK plays a major role in cellular energy metabolism in invertebrates including nematodes. In the present study, we performed the direct immunofluorescence test to determine the immunolocalization of AK in different stages of the life cycle (eggs, larvae, and adult worms) of Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides. Our results indicated variable levels of expression of AK in different stages. Moreover, strong fluorescence was observed in cleaving eggs than in dormant eggs. The highest activity of the enzyme was observed in the fully developed eggs. This may be due to high expression of AK in embryonic development, which is associated with increased energy demand due to cleavage and cellular differentiation. Surprisingly, expression of AK is significantly higher in the middle part and posterior end compared to anterior end of the larvae. In addition, AK is highly concentrated in cellular and metabolically active parts of the body such as hypodermis, muscle, intestine, ovaries, oviducts, and uterus, while it is absent in noncellular areas like cuticle. The present study revealed the presence of AK in T. canis, A. lumbricoides, and T. vitulorum and that it plays a major role in energy metabolism of these nematodes. Interestingly, antiserum was prepared against the recombinant T. canis AK and reacts with the native AKs of T. canis, A. lumbricoides, and T. vitulorum. AK levels could vary in relation to maximum potential rates of ATP turnover, oxidative capacity, and energy output. Further studies on subcellular localization of AK in these important helminths provide new information for researchers to develop effective anthelmintics against the parasites of veterinary and of public health importance.

摘要

精氨酸激酶(AK)是磷酸肌酸激酶家族的一员。AK 在包括线虫在内的无脊椎动物的细胞能量代谢中起着重要作用。在本研究中,我们进行了直接免疫荧光试验,以确定 AK 在犬弓首蛔虫、牛弓首蛔虫和猪蛔虫生命周期的不同阶段(卵、幼虫和成虫)中的免疫定位。我们的结果表明 AK 在不同阶段的表达水平不同。此外,在分裂卵中观察到的荧光比休眠卵强。该酶的最高活性出现在完全发育的卵中。这可能是由于 AK 在胚胎发育中的高表达,这与由于分裂和细胞分化而导致的能量需求增加有关。令人惊讶的是,在幼虫的中部和后端表达 AK 的水平明显高于前端。此外,AK 高度集中在身体的细胞和代谢活跃部位,如皮下组织、肌肉、肠、卵巢、输卵管和子宫,而在无细胞区域如角质层中则不存在。本研究表明 AK 存在于犬弓首蛔虫、猪蛔虫和牛弓首蛔虫中,并且在这些线虫的能量代谢中起主要作用。有趣的是,针对重组犬弓首蛔虫 AK 制备的抗血清与犬弓首蛔虫、猪蛔虫和牛弓首蛔虫的天然 AK 反应。AK 水平可能会因最大潜在 ATP 周转率、氧化能力和能量输出而有所不同。对这些重要寄生虫 AK 的亚细胞定位的进一步研究为研究人员提供了新的信息,以开发针对兽医和公共卫生重要寄生虫的有效驱虫药。

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