Galvani Gerónimo L, Fruttero Leonardo L, Coronel María F, Nowicki Susana, Demartini Diogo R, Defferrari Marina S, Postal Melissa, Canavoso Lilián E, Carlini Célia R, Settembrini Beatriz P
Facultad de Ciencias Biomédicas, Universidad Austral, Pilar, Provincia de Buenos Aires, Argentina; Museo Argentino de Ciencias Naturales, Ciudad Autónoma de Buenos Aires, Argentina.
Dpto. Bioquímica Clínica, Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
Biochim Biophys Acta. 2015 Feb;1850(2):255-62. doi: 10.1016/j.bbagen.2014.11.008. Epub 2014 Nov 13.
Triatoma infestans is the main vector of Chagas'disease in Southern Cone countries. In triatomines, symptoms suggesting neurotoxicity were observed after treatment with Jaburetox (Jbtx), the entomotoxic peptide obtained from jackbean urease. Here, we study its effect in the central nervous system (CNS) of this species.
Immunohistochemistry, Western blots, immunoprecipitation, two-dimensional electrophoresis, tandem mass spectrometry and enzymatic assays were performed.
Anti-Jbtx antibody labeled somata of the antennal lobe only in Jbtx-treated insects. Western blot assays of nervous tissue using the same antibody reacted with a 61kDa protein band only in peptide-injected insects. Combination of immunoprecipitation, two-dimensional electrophoresis and tandem mass spectrometry identified UDP-N-acetylglucosamine pyrophosphorylase (UDP-GlcNAcP) as a molecular target for Jbtx. The activity of UDP-GlcNAcP increased significantly in the CNS of Jbtx-treated insects. The effect of Jbtx on the activity of nitric oxide synthase (NOS) and NO production was investigated as NO is a recognized messenger molecule in the CNS of T. infestans. NOS activity and NO levels decreased significantly in CNS homogenates of Jbtx-treated insects.
UDP-GlcNAcP is a molecular target of Jbtx. Jbtx impaired the activity of T. infestans nitrergic system, which may be related with early behavioral effects.
We report that the CNS of Triatoma infestans is a target for the entomotoxic peptide and propose that a specific area of the brain is involved. Besides potentially providing tools for control strategies of Chagas' disease vectors our data may be relevant in various fields of research as insect physiology, neurobiology and protein function.
克氏锥蝽是南锥体国家恰加斯病的主要传播媒介。在用从刀豆脲酶中获得的昆虫毒性肽Jaburetox(Jbtx)处理后,在锥蝽中观察到了提示神经毒性的症状。在此,我们研究其在该物种中枢神经系统(CNS)中的作用。
进行了免疫组织化学、蛋白质印迹、免疫沉淀、二维电泳、串联质谱和酶活性测定。
抗Jbtx抗体仅在经Jbtx处理的昆虫中标记触角叶的神经元胞体。使用相同抗体对神经组织进行的蛋白质印迹分析仅在注射肽的昆虫中与一条61kDa的蛋白带发生反应。免疫沉淀、二维电泳和串联质谱相结合鉴定出UDP-N-乙酰葡糖胺焦磷酸化酶(UDP-GlcNAcP)是Jbtx的分子靶点。在经Jbtx处理的昆虫的中枢神经系统中,UDP-GlcNAcP的活性显著增加。由于一氧化氮(NO)是克氏锥蝽中枢神经系统中公认的信使分子,因此研究了Jbtx对一氧化氮合酶(NOS)活性和NO产生的影响。在经Jbtx处理的昆虫的中枢神经系统匀浆中,NOS活性和NO水平显著降低。
UDP-GlcNAcP是Jbtx的分子靶点。Jbtx损害了克氏锥蝽的硝化能系统的活性,这可能与早期行为效应有关。
我们报告克氏锥蝽的中枢神经系统是这种昆虫毒性肽的作用靶点,并提出大脑的一个特定区域与之相关。除了可能为恰加斯病传播媒介的控制策略提供工具外,我们的数据可能在昆虫生理学、神经生物学和蛋白质功能等各个研究领域具有相关性。