. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. FERTILIZATION ENVELOPE ASSEMBLY 349. Figure 1. Phase microscopy of Strongylocentrotus piirpiiranix eggs activated in normal (A. E) and Na* depleted SWs (B-D and F-H). A. E: Normal Na+-SW. B, F: KCl-substituted-SW. C, G: Tns-substituted- SW. D, H: ChCl-substituted-SW. A-D: 1 min postactivation. E-H: 30 min. These micrographs were taken focusing on the fertilization envelopes, fe = fertilization envelope; PVS = perivitelline space. Scale bar = 50 ^m. (Fig. 4). FPs were collected at 10 min postactivation from eggs pooled from s
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. FERTILIZATION ENVELOPE ASSEMBLY 349. Figure 1. Phase microscopy of Strongylocentrotus piirpiiranix eggs activated in normal (A. E) and Na* depleted SWs (B-D and F-H). A. E: Normal Na+-SW. B, F: KCl-substituted-SW. C, G: Tns-substituted- SW. D, H: ChCl-substituted-SW. A-D: 1 min postactivation. E-H: 30 min. These micrographs were taken focusing on the fertilization envelopes, fe = fertilization envelope; PVS = perivitelline space. Scale bar = 50 ^m. (Fig. 4). FPs were collected at 10 min postactivation from eggs pooled from several females. The FPs of the normal and K+ -substituted SW eggs had ± Mg and ± Mg (Mean ± ) of protein/ml FP, respectively. They were not significantly different. However, the FPs of Tris- and ChCl-substituted SWs contained 162. 1 ± Mg and ± ^g- respectively. This 7- to 8-fold in- crease over normal and KC1 was highly significant (P < ). Control assays demonstrated no significant difference (95% confidence level) in TCA-precipitable BSA between normal and low Na+ SWs. Therefore, the various SWs had no adverse effects on the Lowry assay. In addition, supernatant protein from unactivated eggs in DMSO was measured and found to contribute little to the total ( ~ 1 .4 h see also Green et ai, 1990). ft- 1 ,3-glucanase secretion Glucanase activity was measured (in normal and ChCl SWs) by the amount of glucose hydrolyzed from the /8- 1,3-glucan polysaccharide laminarin by egg-derived-glu- canase in the FP (Fig. 5). Aliquots of FP of experimental and control groups were taken at 10 min postactivation. The glucose measurements from ChCl and normal SW were ± and ± ^moles glucose per ml of FP, respectively. Approximately 75% more glucanase activity was found in the FP from the ChCl eggs. This difference was significant (P < ). Control incubations of glucose were assayed in normal and ChCl-substituted SWs, an
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