. The Biological bulletin. Biology; Zoology; Marine biology. 106 T. C. JEGLA AND J. D. 00 50-. " I 2 20-HYDR0XYECDYS0NE(ng) Figure 1. Response of first stage Lunulas larvae to doses of 20-hydroxyecdysone in the range of pg to 20 ng. The larvae were 15 to 16 days of age in a molt cycle of 23 days. ful injections. Larvae were held on the pipette tip for about 20 sec after injection to insure entry of the entire injection volume into the hemocoele, then transferred to a labeled culture bowl. Larvae were discarded if they were injected with an excess of
. The Biological bulletin. Biology; Zoology; Marine biology. 106 T. C. JEGLA AND J. D. 00 50-. " I 2 20-HYDR0XYECDYS0NE(ng) Figure 1. Response of first stage Lunulas larvae to doses of 20-hydroxyecdysone in the range of pg to 20 ng. The larvae were 15 to 16 days of age in a molt cycle of 23 days. ful injections. Larvae were held on the pipette tip for about 20 sec after injection to insure entry of the entire injection volume into the hemocoele, then transferred to a labeled culture bowl. Larvae were discarded if they were injected with an excess of more than an estimated 5% of the experimental solution. Routinely, larvae were given one injection of /A of test solution (equals about 4% of the larval volume). Although they will tolerate /x\, considerable swelling does occur with the larger volume. The experiment was stopped 5 to 10 days after injection, depending on age of the larvae at time of injection. All animals were inspected for molting (actually throughout the experiment) or production of a new cuticle. The dorsal prosoma of ttnmolted larvae was dissected with watchmaker's forceps under the stereo- scopic microscope and a new cuticle of about 1 /im thickness was easily detected. Early apolysis was not reliably detected in gross dissection, therefore was not included as an effect. Animals molting before 96 hr post-injection time were discarded. Experimental effects were thus based solely on new cuticle forma- tion and molting after a set period of time. The number of larvae affected among the survivors (usually 100%) was converted to a per cent effect: for experimental groups this was corrected for the corresponding control effect by the following method: if 100 - % Control Effect = Y; and 100 - % Experimental Effect = X; then X -r- Y = % Effect, corrected. Radioimmune assay technique An antibody to 20-hydroxyecdysone was kindly provided by Professor G. R. Wyatt of Queens University, Kingston, Ontario, Canada
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