Endo Y, Ohnishi A, Hayakawa Y
Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo, Kyoto, Japan
J Insect Physiol. 1998 Sep;44(9):859-866. doi: 10.1016/s0022-1910(98)00017-1.
Growth-blocking peptide (GBP) has been purified for the first time from the haemolymph of the host armyworm Pseudaletia separata whose growth is inhibited and shows developmental arrest in the last larval instar stage when parasitized by the parasitoid wasp Cotesia kariyai. GBP naturally occurs in the haemolymph of lepidopteran larvae but its concentration is very low during the last larval instar in comparison with that in the penultimate larval instar. However, by 24h after parasitization or polydnavirus (PdV)-infection on day 0 of the last larval instar, a four-fold increase in GBP level, compared with synchronous non-parasitized control larvae, is observed. Although Northern blot analysis indicates that GBP mRNA is transcribed in brain-nerve cord and fat body, plasma GBP is likely to be secreted mainly from fat body because the GBP mRNA level is approximately 100-fold higher in fat body than that in brain-nerve cord. RT-PCR analysis demonstrates the constant expression of GBP mRNA in both parasitized (or PdV-infected) and non-parasitized larval fat body, which suggests that parasitism does not influence transcriptional level, but might influence post-transcriptional level to elevate plasma GBP concentration. This interpretation was supported by estimating GBP precursor levels in fat body of PdV-infected and non-infected larvae. Virus infection appears to elevate the GBP precursor levels in fat body to about six times greater than that in non-infected last instar larvae by 6h after PdV-injection. The GBP processing enzyme activity that occurs in Golgi body-rich extract of the fat body is increased by about 90% after parasitization or PdV-injection.
生长阻滞肽(GBP)首次从宿主粘虫(Pseudaletia separata)的血淋巴中纯化得到。当被寄生蜂梨小食心虫(Cotesia kariyai)寄生时,宿主粘虫的生长受到抑制,并在最后一龄幼虫阶段出现发育停滞。GBP天然存在于鳞翅目幼虫的血淋巴中,但与倒数第二龄幼虫相比,其在最后一龄幼虫期的浓度非常低。然而,在最后一龄幼虫第0天被寄生或感染多角体病毒(PdV)后24小时,与同步未被寄生的对照幼虫相比,GBP水平增加了四倍。虽然Northern印迹分析表明GBP mRNA在脑-神经索和脂肪体中转录,但血浆GBP可能主要从脂肪体分泌,因为脂肪体中的GBP mRNA水平比脑-神经索中的高约100倍。RT-PCR分析表明,在被寄生(或感染PdV)和未被寄生的幼虫脂肪体中,GBP mRNA均持续表达,这表明寄生不影响转录水平,但可能影响转录后水平以提高血浆GBP浓度。通过估计感染PdV和未感染的幼虫脂肪体中的GBP前体水平,这一解释得到了支持。病毒感染似乎在注射PdV后6小时将脂肪体中的GBP前体水平提高到比未感染的最后一龄幼虫高约六倍。寄生或注射PdV后,脂肪体中富含高尔基体提取物中的GBP加工酶活性增加了约90%。