Zitnan D, Kingan T G, Kramer S J, Beckage N E
Department of Entomology, University of California, Riverside 92521, USA.
J Comp Neurol. 1995 May 22;356(1):83-100. doi: 10.1002/cne.903560106.
Fifth instar larvae of Manduca sexta that were parasitized by the braconid wasp Cotesia congregata failed to develop after the parasitoid larvae emerged, and these host larvae lingered for 2-3 weeks in a quiescent, nonfeeding state without initiating a larval molt or metamorphosis. This study was focused on the neuroendocrine changes associated with the host's developmental arrest. Immunohistochemical studies suggested that the host brain neurosecretory cells as well as their axon terminals in the corpora cardiaca-corpora allata complex accumulated multiple neuropeptides. The extent of accumulation in cells and axons increased with time, so that hosts examined 7-14 days after the wasps emerged showed the most intense staining with antibodies against prothoracicotropic hormone, bombyxin, allatotropin, allatostatin, diuretic hormone, eclosion hormone, proctolin, and FMRFamide. Increased levels of prothoracicotropic hormone and FMRFamide-like peptides in the brains of parasitized larvae were confirmed using Western blots and enzyme-linked immunosorbent assay (ELISA), respectively. Starvation of the unparasitized larvae induced some accumulation of the neuropeptides; however, the intensity of staining and number of immunopositive cells and axons were in most cases clearly higher in the parasitized larvae. Our results suggest that accumulation of the neuropeptides is associated with developmental arrest of parasitized larvae. Because a similar developmental arrest occurs in a wide range of parasitized insects, our findings may have relevance for many other species. Moreover, these data illustrate the potential value of using parasitized M. sexta larvae as a model for studying the mechanisms governing the rates of neuropeptide expression, processing, packaging, and release, as well as providing a rich source of neuropeptides, thus facilitating their isolation and characterization.
被茧蜂科黄蜂聚集茧蜂寄生的烟草天蛾五龄幼虫,在寄生蜂幼虫羽化后无法发育,这些寄主幼虫会在静止、不进食的状态下徘徊2至3周,既不开始幼虫蜕皮也不变态。本研究聚焦于与寄主发育停滞相关的神经内分泌变化。免疫组织化学研究表明,寄主脑内神经分泌细胞及其在咽侧体 - 心侧体复合体中的轴突终末积累了多种神经肽。细胞和轴突中的积累程度随时间增加,因此在黄蜂羽化后7至14天检查的寄主,用抗促前胸腺激素、家蚕素、促咽侧体素、咽侧体抑制素、利尿激素、羽化激素、促肠肌动素和FMRF酰胺的抗体染色最为强烈。分别使用蛋白质免疫印迹法和酶联免疫吸附测定(ELISA)证实了被寄生幼虫脑内促前胸腺激素和FMRF酰胺样肽水平升高。未被寄生的幼虫饥饿会诱导一些神经肽的积累;然而,在大多数情况下,被寄生幼虫的染色强度以及免疫阳性细胞和轴突的数量明显更高。我们的结果表明,神经肽的积累与被寄生幼虫的发育停滞有关。由于在多种被寄生昆虫中都会发生类似的发育停滞,我们的发现可能与许多其他物种相关。此外,这些数据说明了利用被寄生的烟草天蛾幼虫作为模型来研究神经肽表达、加工、包装和释放速率调控机制的潜在价值,同时提供了丰富的神经肽来源,从而便于对其进行分离和鉴定。