The conserved role of the AKT/GSK3 axis in cell survival and glycogen metabolism in Rhipicephalus (Boophilus) microplus embryo tick cell line BME26
The conserved role of the AKT/GSK3 axis in cell survival and glycogen metabolism in Rhipicephalus (Boophilus) microplus embryo tick cell line BME26
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DOI：
10.1016/j.bbagen.2012.12.016
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发表时间：
2013-03-01
影响因子：
3
通讯作者：
Logullo, Carlos
中科院分区：
文献类型：
--
作者：
de Abreu, Leonardo Araujo;Calixto, Christiano;Logullo, Carlos
背景：蜱的胚胎发生是在严格控制的条件下进行的代谢密集过程，其组成成分知之甚少。方法：为了研究AKT（蛋白激酶B）在糖原代谢和细胞活力中的作用，在RNA干扰（RNAi）和化学抑制实验的同时，对微加棘头猪（Rhipicephalus microplus）进行糖原测定、鉴定和克隆。结果：10-DEBC化学抑制AKT可降低糖原含量，而阿斯特保龙抑制GSK3则相反。RmAKT ORF全长1584 bp，编码60.1 kDa的多肽链。系统发育和序列分析显示脊椎动物和蜱类之间存在显著差异。AKT或GSK3敲除的细胞靶转录物水平均降低70%，但AKT的降低也降低了糖原含量、细胞活力和细胞膜通透性。然而，GSK3的减少促进了糖原含量的增加。此外，GSK3抑制或基因沉默对暴露于紫外线后的BME26活性均有保护作用。AKT和GSK3在胚胎发育过程中广泛表达。综上所述，我们的数据支持AKT和GSK3的拮抗作用，并强烈表明这种信号轴在蜱胚胎中是保守的，AKT位于GSK3的上游。一般意义：AKT/GSK3轴在蜱虫中以一种整合糖原代谢和细胞存活的方式保守，并表现出系统发育差异，这可能对开发新的控制方法很重要。(C) 2013 Elsevier B.V.版权所有
Background: Tick embryogenesis is a metabolically intensive process developed under tightly controlled conditions and whose components are poorly understood.Methods: In order to characterize the role of AKT (protein kinase B) in glycogen metabolism and cell viability, glycogen determination, identification and cloning of an AKT from Rhipicephalus microplus were carried out, in parallel with experiments using RNA interference (RNAi) and chemical inhibition.Results: A decrease in glycogen content was observed when AKT was chemically inhibited by 10-DEBC treatment, while GSK3 inhibition by alsterpaullone had an opposing effect. RmAKT ORF is 1584-bp long and encodes a polypeptide chain of 60.1 kDa. Phylogenetic and sequence analyses showed significant differences between vertebrate and tick AKTs. Either AKT or GSK3 knocked down cells showed a 70% reduction in target transcript levels, but decrease in AKT also reduced glycogen content, cell viability and altered cell membrane permeability. However, the GSK3 reduction promoted an increase in glycogen content. Additionally, either GSK3 inhibition or gene silencing had a protective effect on BME26 viability after exposure to ultraviolet radiation. R. microplus AKT and GSK3 were widely expressed during embryo development. Taken together, our data support an antagonistic role for AKT and GSK3, and strongly suggest that such a signaling axis is conserved in tick embryos, with AKT located upstream of GSK3.General significance: The AKT/GSK3 axis is conserved in tick in a way that integrates glycogen metabolism and cell survival, and exhibits phylogenic differences that could be important for the development of novel control methods. (C) 2013 Elsevier B.V. All rights reserved.