Hayes D K, Bird T G, Mills G D, Frankoff H
Beltsville Agricultural Research Center, U.S. Department of Agriculture, MD 20705.
Chronobiol Int. 1990;7(5-6):413-8. doi: 10.3109/07420529009059152.
Trehalose levels were determined over two 24 hr spans in groups of face fly adults 3-4 days after emergence from the puparium. Face fly pupae were placed in rearing chambers at 27 degrees C in a staggered light-dark regimen, LD 16:8, so that at a given clock hour, samples could be obtained at several different hours after lights on (HALO). Trehalose was determined in hemolymph collected from a puncture in the intersegmental membrane of the abdomen. Treated hemolymph samples were passed through a Bio-Rad Amino 5-S disaccharide column and a Waters 410 refractive index detector was used to differentiate among sugars. The circadian acrophase derived by cosinor analysis in hemolymph trehalose (when the values were 25.49 and 26.86 micrograms/microliters on the first and second days respectively) occurred at -226 degrees (ca 15 HALO) and the bathyphase at 24 HALO. The mesor = 11.82 micrograms/microliters trehalose, the amplitude = 8.57 micrograms/microliters trehalose and the P-value for presence of a rhythm was 0.003. Based on these data, differences between control and test flies in a bioassay of hypertrehalosemic activity would be most easily observed at 0-8 HALO, while exogenous hypotrehalosemic activity would be best assayed at 12-20 HALO.
在从蛹羽化后3 - 4天的面蝇成虫组中,在两个24小时时间段内测定海藻糖水平。将面蝇蛹置于27摄氏度的饲养室中,采用交错的明暗周期,即LD 16:8,这样在给定的时钟时间,可以在光照后几个不同的小时(HALO)获取样本。从腹部节间膜穿刺收集的血淋巴中测定海藻糖。处理后的血淋巴样本通过Bio - Rad Amino 5 - S二糖柱,并使用Waters 410折光率检测器区分不同的糖类。通过余弦分析得出的血淋巴海藻糖的昼夜节律峰相位(第一天和第二天的值分别为25.49和26.86微克/微升时)出现在 - 226度(约15 HALO),谷相位出现在24 HALO。中值 = 11.82微克/微升海藻糖,振幅 = 8.57微克/微升海藻糖,节律存在的P值为0.003。基于这些数据,在高海藻糖血症活性生物测定中,对照蝇和测试蝇之间的差异在0 - 8 HALO时最容易观察到,而外源性低海藻糖血症活性在12 - 20 HALO时测定效果最佳。