Dotson E M, Connat J L, Diehl P A
Institut de Zoologie, Université de Neuchâtel, Chantemerle, Neuchâtel, Switzerland.
Arch Insect Biochem Physiol. 1993;23(2):67-78. doi: 10.1002/arch.940230203.
The fate of [3H]-ecdysone ([3H]-E) was investigated in hanging drop cultures of embryos and larvae of the tick Ornithodoros moubata using HPLC. The hormone was metabolized more slowly during described periods of increasing endogenous ecdysteroid (ES) titers than during periods of low titers except for young embryos. Three different classes of metabolites were produced: 1) apolar products (AP) corresponding to C-22 fatty acid ester conjugates of E and, in some cases, of 20-hydroxyecdysone (20E), 2) unidentified polar products (PP) more polar than E, one peak of which had the same retention times as 20,26-dihydroxyecdysone, and finally, 3) 20E verified by comigration of cold standards on RP-18 and silica columns. Hydroxylation of E to 20E first became evident in cultures of 2 day old embryos and was present in all cultures of older animals. Highest production of free 20E occurred during increasing endogenous ES titers in embryos and during the ES peak in larvae. Conjugation of E to AP occurred in all stages investigated, but was more pronounced during periods of low endogenous ES titers, and may correspond to a detoxification mechanism. In contrast, PP were produced during high 20E production in embryos and during periods of high and decreasing endogenous titers in larvae.
利用高效液相色谱法(HPLC)研究了放射性标记的蜕皮激素([³H]-E)在多乳突钝缘蜱胚胎和幼虫悬滴培养物中的代谢情况。除了幼胚外,在所描述的内源性蜕皮甾体(ES)滴度升高期间,该激素的代谢速度比滴度低时更慢。产生了三类不同的代谢产物:1)非极性产物(AP),对应于E以及某些情况下20-羟基蜕皮激素(20E)的C-22脂肪酸酯共轭物;2)比E极性更强的未鉴定极性产物(PP),其中一个峰的保留时间与20,26-二羟基蜕皮激素相同;最后,3)通过冷标准品在RP-18和硅胶柱上的共迁移验证的20E。E羟基化为20E首先在2日龄胚胎培养物中明显出现,并存在于所有老龄动物培养物中。游离20E的最高产量出现在胚胎内源性ES滴度升高期间以及幼虫的ES峰值期间。E与AP的共轭发生在所有研究阶段,但在内源性ES滴度较低期间更为明显,可能对应一种解毒机制。相反,PP在胚胎中20E产量高时以及幼虫内源性滴度高和下降期间产生。