Range Sebastian, Oeh Uwe, Lorenz Matthias W, Etzel Winfried, Nauen Ralf, Hoffmann Klaus H
Animal Ecology I, University of Bayreuth, D-95440 Bayreuth, Germany.
Comp Biochem Physiol B Biochem Mol Biol. 2002 May;132(1):191-202. doi: 10.1016/s1096-4959(01)00522-x.
The in vitro production of juvenile hormones (JH) was investigated by using corpora allata (CA) of larvae and corpora cardiaca-corpora allata (CC-CA) complexes of adult females of the fall armyworm Spodoptera frugiperda. In female moths, JH release was high compared to that in 5th and 6th instar larvae. Concentrations of 0.11-0.12 mM methionine, 180-200 mM Na(+), 5.8-8.3 mM K(+), 10-50 mM Ca(2+) and a pH range of 5.7-6.3 yielded optimal incorporation of L-[methyl-(3)H] methionine in vitro by CC-CA complexes. The highest hourly incorporation occurred during a 9-h incubation period following a 1.5-h lag-phase. JH release from CC-CA complexes of adult females was shown to be age-dependent with a peak value on day 2 (approx. 4 pmol h(-1) CA(-1)). By a combination of reversed phase (RP)- and normal phase (NP)-high performance liquid chromatography (HPLC), two major labelled products released by the complex were separated. One compound co-migrated with chemically synthesized JH II diol, the second compound with JH III diol. Only traces of JH II and III could be detected in some samples. Gland extracts also contained both the major radiolabelled products. Double labelling experiments using [3H]methionine and [14C]acetate confirmed their de novo synthesis in CC-CA complexes of female moths. The nature of chemically synthesized reference JH III diol was proved by LC-MS (ESI mass spectrometry) and 1H-NMR (nuclear magnetic resonance spectroscopy).
利用草地贪夜蛾幼虫的咽侧体（CA）和成年雌虫的心侧体 - 咽侧体（CC - CA）复合体研究了保幼激素（JH）的体外产生情况。在雌蛾中，与5龄和6龄幼虫相比，JH释放量较高。0.11 - 0.12 mM的甲硫氨酸、180 - 200 mM的Na⁺、5.8 - 8.3 mM的K⁺、10 - 50 mM的Ca²⁺以及5.7 - 6.3的pH范围可使CC - CA复合体在体外对L - [甲基 - ³H]甲硫氨酸的掺入达到最佳效果。在1.5小时的延迟期后的9小时孵育期内，每小时的掺入量最高。成年雌虫的CC - CA复合体释放的JH显示出年龄依赖性，在第2天达到峰值（约4 pmol h⁻¹ CA⁻¹）。通过反相（RP）和正相（NP）高效液相色谱（HPLC）相结合的方法，分离出了复合体释放的两种主要标记产物。一种化合物与化学合成的JH II二醇共迁移，第二种化合物与JH III二醇共迁移。在一些样品中仅能检测到痕量的JH II和III。腺体提取物中也含有这两种主要的放射性标记产物。使用[³H]甲硫氨酸和[¹⁴C]乙酸盐的双重标记实验证实了它们在雌蛾的CC - CA复合体中是从头合成的。通过液相色谱 - 质谱联用（ESI质谱）和¹H - 核磁共振光谱（NMR）证明了化学合成的参考JH III二醇的性质。
