Li Yiping, Hernandez-Martinez Salvador, Unnithan Gopalan C, Feyereisen René, Noriega Fernando G
Department of Biochemistry and Molecular Biophysics, and Center for Insect Science, University of Arizona, Tucson, AZ 85721, USA.
Insect Biochem Mol Biol. 2003 Dec;33(12):1307-15. doi: 10.1016/j.ibmb.2003.07.003.
The synthesis of juvenile hormone III (JH III) by the isolated corpora allata (CA) of Aedes aegypti adult female was studied using an in vitro radiochemical assay. We dissected the corpora allata-corpora cardiaca (CA-CC) complex attached to a piece of aorta. The complex was left connected to the intact head capsule to facilitate the visualization and transfer of the glands. A linear increase in the cumulative amount of biosynthesized JH III was found for at least the first 6 h of incubation; approximately 45% of the synthesized JH III was present in the medium. There was a dependence of JH III synthesis on exogenous methionine supply. Using reversed phase high performance liquid chromatography two major labeled products biosynthesized by the CA were separated. They co-migrated with JH III and methyl farnesoate (MF). The identity of the biosynthesized JH III was confirmed by gas chromatography-mass spectrometry. JH III synthesis was only 2.0 fmol/pair gland/h immediately after adult emergence, but increased to 32.6 fmol/ pair gland/h 18 h later in sugar-fed females. Two days after emergence, the CA biosynthetic activity slowly started to decrease, and reached values of around 5.3 fmol/pair gland/h by one week after emergence. Synthesis of JH was similar from either sugar-fed females mated or unmated. A blood meal resulted in a decrease of JH III synthesis in CA from mated females by 12 h after feeding and from virgin females by 24 h after feeding. JH III biosynthesis remained low for at least 96 h in mated females, but was back to higher levels 72 h after feeding in virgin females. Rates of JH III biosynthesis closely reflected the hemolymph levels of JH III both after emergence and after a blood meal described by Shapiro et al. (1986). The activity of the CA in Aedes aegypti females seems to be regulated by developmental changes and nutritional signals, and to be independent of mating stimulus.
利用体外放射化学分析法,研究了埃及伊蚊成年雌蚊分离的咽侧体(CA)合成保幼激素III(JH III)的情况。我们解剖了附着在一段主动脉上的咽侧体 - 心侧体(CA - CC)复合体。该复合体保持与完整的头壳相连,以利于腺体的观察和转移。在至少最初6小时的孵育过程中,生物合成的JH III累积量呈线性增加;约45%合成的JH III存在于培养基中。JH III的合成依赖于外源性甲硫氨酸的供应。使用反相高效液相色谱法,分离出了CA生物合成的两种主要标记产物。它们与JH III和法尼醇甲酯(MF)共迁移。通过气相色谱 - 质谱法确认了生物合成的JH III的身份。刚羽化后,JH III的合成仅为2.0 fmol/对腺体/小时,但在取食糖水的雌蚊中,18小时后增加到32.6 fmol/对腺体/小时。羽化两天后,CA的生物合成活性开始缓慢下降,到羽化一周后达到约5.3 fmol/对腺体/小时的值。取食糖水的已交配或未交配雌蚊的JH合成情况相似。一次血餐导致已交配雌蚊的CA中JH III合成在进食后12小时下降,未交配雌蚊在进食后24小时下降。在已交配雌蚊中,JH III生物合成至少96小时保持在低水平,但未交配雌蚊在进食后72小时恢复到较高水平。JH III生物合成速率在羽化后和血餐之后都密切反映了Shapiro等人(1986年)描述的JH III的血淋巴水平。埃及伊蚊雌蚊的CA活性似乎受发育变化和营养信号调节,且与交配刺激无关。