Ohnishi A, Hayakawa Y, Matsuda Y, Kwon K W, Takahashi T A, Sekiguchi S
Biochemical Laboratory, Hokkaido University, Sapporo, Japan.
Insect Biochem Mol Biol. 1995 Dec;25(10):1121-7. doi: 10.1016/0965-1748(95)00054-2.
Growth-blocking peptide (GBP) has been isolated for the first time from the haemolymph of the host armyworm Pseudaletia separata whose development was halted in the last larval instar stage by parasitization with the parasitoid wasp Cotesia kariyai. Recent studies demonstrated that GBP not only exists in the plasma (haemolymph without cells) of parasitized last instar larvae, but also in the plasma of nonparasitized penultimate (5th) instar larvae. Monoclonal antibodies were prepared to measure the titers of GBP in nonparasitized and parasitized larval plasma. One of three monoclonal antibodies raised against GBP, which is the most specific for GBP, was used to quantify the concentration of plasma GBP. As this antibody recognized two plasma peptides other than GBP in crude plasma fractions, each plasma peptide fraction was separated by a reversed phase HPLC, and then plasma GBP level was measured by ELISA. The highest level of plasma GBP detected on Day 0 of the penultimate instar larvae was gradually decreased throughout the larval growth except for the temporary increase on Day 0 of last larval instar. After parasitization on Day 0 of last larval instar, two peaks of plasma GBP titer were detected during the last larval instar, one day and six days after parasitization. This characteristic increase and decrease in plasma GBP level was also observed by transferring last instar larvae of the armyworm from 25 to 10 degrees C, as a result of which larvae delayed pupation by more than 15 days. From these results, it is reasonable to propose that plasma GBP in lepidopteran larvae might control certain upstream steps in a cascade of events leading to pupation; thus, an elevated level of plasma GBP interferes with normal metamorphosis from larvae to pupae.
生长阻滞肽(GBP)首次从宿主粘虫(Pseudaletia separata)的血淋巴中分离出来,该宿主粘虫在最后一龄幼虫阶段因被寄生蜂梨形齿腿姬蜂(Cotesia kariyai)寄生而发育停滞。最近的研究表明,GBP不仅存在于被寄生的最后一龄幼虫的血浆(无细胞血淋巴)中,也存在于未被寄生的倒数第二(第五)龄幼虫的血浆中。制备了单克隆抗体来测量未被寄生和被寄生幼虫血浆中GBP的滴度。针对GBP产生的三种单克隆抗体中,对GBP最具特异性的一种用于定量血浆GBP的浓度。由于该抗体在粗血浆组分中识别出除GBP之外的两种血浆肽,每个血浆肽组分通过反相高效液相色谱法进行分离,然后通过酶联免疫吸附测定法测量血浆GBP水平。在倒数第二龄幼虫的第0天检测到的血浆GBP最高水平在整个幼虫生长过程中逐渐下降,除了在最后一龄幼虫的第0天出现暂时升高。在最后一龄幼虫的第0天被寄生后,在最后一龄幼虫期间检测到血浆GBP滴度出现两个峰值,分别在寄生后的第1天和第6天。通过将粘虫的最后一龄幼虫从25℃转移到10℃也观察到了血浆GBP水平的这种特征性升高和降低,结果幼虫化蛹延迟超过15天。从这些结果来看,有理由推测鳞翅目幼虫血浆中的GBP可能控制导致化蛹的一系列事件中的某些上游步骤;因此,血浆GBP水平升高会干扰从幼虫到蛹的正常变态。