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刻点血蜱和微小扇头蜱中自噬相关基因的相对转录

Relative transcription of autophagy-related genes in Amblyomma sculptum and Rhipicephalus microplus ticks.

作者信息

Moura-Martiniano Nicole O, Machado-Ferreira Erik, Gazêta Gilberto S, Soares Carlos Augusto Gomes

机构信息

Laboratório de Genética Molecular de Eucariontes e Simbiontes, Departamento de Genética, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

Laboratório de Referência Nacional em Vetores das Riquetsioses, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.

出版信息

Exp Appl Acarol. 2017 Dec;73(3-4):401-428. doi: 10.1007/s10493-017-0193-z. Epub 2017 Nov 27.

DOI:10.1007/s10493-017-0193-z
PMID:29181673
Abstract

Ticks endure stressful off-host periods and perform as vectors of a diversity of infectious agents, thus engaging pathways that expectedly demand for autophagy. Little is known of ticks' autophagy, a conserved eukaryotic machinery assisting in homeostasis processes that also participates in tissue-dependent metabolic functions. Here, the autophagy-related ATG4 (autophagin-1), ATG6 (beclin-1) and ATG8 (LC3) mRNAs from the human diseases vector Amblyomma sculptum and the cattle-tick Rhipicephalus microplus were identified. Comparative qPCR quantifications evidenced different transcriptional status for the ATG genes in the salivary glands (SG), ovaries and intestines of actively feeding ticks. These ATGs had increased relative transcription under nutrient-deprivation, as determined by validation tests with R. microplus embryo-derivative cells BME26 and A. sculptum SG explants incubations in HBSS. Starvation lead to 4-31.8× and ~ 60-500× increments on the ATGs mRNA loads in BME26 and A. sculptum SG explants, respectively. PI3K inhibitor 3MA treatment also affected ATGs expression in BME26. Some ATGs were more transcribed in the SGs than in the ovaries of cattle-ticks. Amblyomma sculptum/R. microplus interspecific comparisons showed that ATG4 and ATG6 were 0.18× less expressed in A. sculptum SGs, but ~ 10-100× more expressed in their ovaries when compared to R. microplus organs. ATG4 and ATG8a transcript loads were ~ 120× and ~ 40× higher, respectively, in A. sculptum intestines when compared to cattle-ticks of similar weight category. ATGs expression in A. sculptum intestines increased with tick weight, indicating Atgs contribution to intracellular blood digestion. Possible roles of the autophagy machinery and their organ-specific expression profile on vector biology are discussed.

摘要

蜱虫会经历压力较大的非寄生期,并作为多种感染因子的传播媒介,因此会启动一些预计需要自噬的途径。人们对蜱虫的自噬了解甚少,自噬是一种保守的真核机制,有助于体内平衡过程,也参与组织依赖性代谢功能。在此,我们鉴定了来自人类疾病传播媒介刻点钝缘蜱和微小牛蜱的自噬相关基因ATG4(自噬相关蛋白1)、ATG6(贝克林1)和ATG8(微管相关蛋白轻链3)的mRNA。比较定量PCR结果表明,在饱血蜱虫的唾液腺、卵巢和肠道中,ATG基因的转录状态不同。通过用微小牛蜱胚胎衍生细胞BME26和刻点钝缘蜱唾液腺外植体在无钙镁平衡盐溶液中孵育进行验证试验,确定这些自噬相关基因在营养剥夺条件下相对转录增加。饥饿分别导致BME26和刻点钝缘蜱唾液腺外植体中自噬相关基因mRNA含量增加4至31.8倍和约60至500倍。PI3K抑制剂3MA处理也影响BME26中自噬相关基因的表达。一些自噬相关基因在微小牛蜱的唾液腺中的转录水平高于卵巢。刻点钝缘蜱/微小牛蜱种间比较表明,与微小牛蜱的器官相比,ATG4和ATG6在刻点钝缘蜱唾液腺中的表达量低0.18倍,但在其卵巢中的表达量高约10至100倍。与体重相近的微小牛蜱相比,刻点钝缘蜱肠道中ATG4和ATG8a的转录量分别高约120倍和约40倍。刻点钝缘蜱肠道中自噬相关基因的表达随蜱虫体重增加而增加,表明自噬相关基因有助于细胞内血液消化。本文讨论了自噬机制及其器官特异性表达谱在蜱虫生物学中的可能作用。

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