. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. LOBSTER ECDYSTEROID METABOLISM 477 100 >s •o o LjJ -C a ^ o E 0) 0-. 12 24 36 48 60 72 84 96 Time After Injection (h) Figure 1. Changes in the hemolymph level of injected ['H]-ecdysone (E) as a function of time and molt stage. [3H]-ecdysone was injected at time zero. Concentration of labeled ecdysone in the hemolymph ot lob- sters is expressed as a percentage of the total [3H]-ecdysteroids, and de- termined by methanolic extraction of hemolymph samples followed by scintillation counting of reverse phase-HPLC fractions. Sa
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. LOBSTER ECDYSTEROID METABOLISM 477 100 >s •o o LjJ -C a ^ o E 0) 0-. 12 24 36 48 60 72 84 96 Time After Injection (h) Figure 1. Changes in the hemolymph level of injected ['H]-ecdysone (E) as a function of time and molt stage. [3H]-ecdysone was injected at time zero. Concentration of labeled ecdysone in the hemolymph ot lob- sters is expressed as a percentage of the total [3H]-ecdysteroids, and de- termined by methanolic extraction of hemolymph samples followed by scintillation counting of reverse phase-HPLC fractions. Samples were separated with gradient systems #1 or #2 (see text) with either (or both, for some samples) methanol or acetonitrile as the solvent. Molt stages A. C, D0, D,, and D2 refer to the morphological designations of Aiken (1973). Sample sizes were as given for Table I with the addition of n = 3 for stage A. Standard deviation bars are omitted for clarity. (1) a 35 min linear gradient of 20-100% methanol in water at ml/min ( min fractions); (2) a linear gradient of 20-100% acetonitrile in 20 mM Tris, pH , at ml/ min ( min fractions collected); or (3) a linear gradient of 20-100% methanol in 20 , pH , at ml/ min ( min fractions collected). In all cases, we employed a Waters HPLC system. Duplicate samples from each fraction were analyzed by scintillation spectrometry. The sum of the radioactive ecdysteroids recovered in the HPLC-fractions was equal to 70-85% of the total, un- fractionated radioactivity. The amount of each [3H]-ec- dysteroid metabolite was expressed as the percentage of the total radioactivity, and the values at each time point were compared statistically by ANOVA and Scheffe tests of arcsine transformed values (Sokal and Rohlf, 1969). Enzymatic hydrolysis The fractions resulting from HPLC that contained ec- dysteroids of greater polarity than 20E are designated "polar ; The polar fractions from individua
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
