. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 18 P. S. GLAS ET Figures 15-18. A time series of Sicyonia inguntis eggs in artificial seawater shows the elevation of the hatching envelope (HE) under light microscopy (a) and labeling of the HE with an oxidase sensitive fluorescent dye, dihydrotetramethylrosamine (b). Figure 15. (a) Normal egg before elevation at 35 min postspawn. Two sperm are visible on the exterior of the egg (arrowhead). Arrow indicates first polar body, (b) No fluorescence is visible in the cortex; however, the sperm are fluorescent (arrowhead). T
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 18 P. S. GLAS ET Figures 15-18. A time series of Sicyonia inguntis eggs in artificial seawater shows the elevation of the hatching envelope (HE) under light microscopy (a) and labeling of the HE with an oxidase sensitive fluorescent dye, dihydrotetramethylrosamine (b). Figure 15. (a) Normal egg before elevation at 35 min postspawn. Two sperm are visible on the exterior of the egg (arrowhead). Arrow indicates first polar body, (b) No fluorescence is visible in the cortex; however, the sperm are fluorescent (arrowhead). The first polar body (arrow) can also be seen. Figure 16. At 47 min postspawn. the HE has begun to lift from the oolemma. (a) The HE (HE) is separating from the oolemma and has extranumerary sperm on its exterior, (b) The cortex can be seen to react with DHTMR. Figure 17. By 60 min postspawn. the HE is fully formed, (a) A normal HE (HE) with the second polar body (pb) visible in the perivitelline space, (b) The HE fluoresces brightly, as does the cortex of the egg. The polar body does not fluoresce. Figure 18. At 100 min postspawn. the two-cell stage can be seen, (a) The cells are visible within the ele\ated HE (HE), (b) The cortex still fluoresces, but the HE does not. Bar equals 100 urn. ability of the HE (Table I). Based on the relative fluores- cent intensity inside vs. outside the PVS at an equato- rial focus, both the control and ATA-treated eggs had fluorescent dextrans within the PVS when incubated with the 4400 KDa sugar. When incubated with the 10,000 KDa dextran, the ATA-treated eggs contained flu- orescent dextrans within the PVS, but eggs in ASW did not. The entry of the dextrans was variable in the ATA- treated eggs with 40,000 and Da dextrans, although the control eggs revealed no fluorescent dextrans within the PVS (Table I). Even though extreme care was used, the presence of high molecular weight fluorescent dextrans in the PVS of ATA-treated eggs may hav
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