Cordeiro Bruno A, Tibúrcio Victor Hugo S, Hallwass Mariana, Paes Hugo C, Ribeiro Bergmann M, Báo Sônia N
Programa de Pós-Graduação em Biologia Molecular, Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, 70910-900, Brazil.
J Invertebr Pathol. 2008 May;98(1):7-19. doi: 10.1016/j.jip.2008.01.001. Epub 2008 Jan 12.
Malpighian tubules constitute the main excretion organ of insects. Infection by egt(-) recombinant AcMNPV baculovirus in lepidopteran larvae promotes early degeneration of these structures, which has been correlated with earlier death of the host. However, no trace of viral infection has been detected in that tissue. We constructed two AgMNPV recombinants with the egfp gene under control of the hsp70 promoter, one being egt(-), and used another two recombinants (one egt(-)) containing the lacZ gene. Morphological alterations in the tubules were analyzed by light and electron microscopies. Bioassays were conducted to compare the pathogenicity of recombinants. Results showed progressive presence of marker proteins and tissue degeneration without signals of infection in the tissue. Morphological and bioassay results showed increased pathogenicity for lacZ-containing recombinants compared to the egfp ones; as for egt(-) viruses, we noted higher intensity and earlier onset of alterations. The absence of infection led us to believe that Malpighian tubules degeneration is provoked initially by the death of tracheal cells attached to the tubules and later, by the death of Malpighian tubule cells themselves. Tubule cell death might be due to oncosis and apoptosis, which may be activated by depletion of energy reserves and by accumulation of marker proteins, respectively. Absence of the egt gene may be leading to a higher energetic expense due to molting, thus aggravating tubule cell death, resulting in faster death of host.
马氏管是昆虫的主要排泄器官。在鳞翅目幼虫中,感染缺乏蜕皮甾体尿苷二磷酸葡萄糖基转移酶(egt)的重组苜蓿银纹夜蛾核型多角体病毒(AcMNPV)杆状病毒会促使这些结构过早退化,这与宿主更早死亡有关。然而,在该组织中未检测到病毒感染的痕迹。我们构建了两个在热休克蛋白70(hsp70)启动子控制下带有增强型绿色荧光蛋白(egfp)基因的苜蓿银纹夜蛾核型多角体病毒(AgMNPV)重组体,其中一个缺乏egt基因,并使用了另外两个含有β-半乳糖苷酶(lacZ)基因的重组体(一个缺乏egt基因)。通过光学显微镜和电子显微镜分析了马氏管的形态变化。进行生物测定以比较重组体的致病性。结果显示标记蛋白逐渐出现且组织退化,但组织中没有感染信号。形态学和生物测定结果表明,与含有egfp基因的重组体相比,含有lacZ基因的重组体致病性增强;对于缺乏egt基因的病毒,我们注意到变化的强度更高且出现更早。未检测到感染使我们认为,马氏管退化最初是由附着在马氏管上的气管细胞死亡引起的,随后是马氏管细胞自身死亡。马氏管细胞死亡可能是由于细胞肿胀和凋亡,分别可能由能量储备耗尽和标记蛋白积累激活。缺乏egt基因可能由于蜕皮导致更高的能量消耗,从而加重马氏管细胞死亡,导致宿主更快死亡。