. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 102 P. L. HERTZLER. Figure 2. Effects of cytochalasin D (CD) treatments. For inhibition of polar body 2 (PB2) formation, eggs were transferred at 40 min post-spawning (PS) to 1 /n/W ( mg/1) CD in artificial seawater containing 50 ;uM (ATA-ASW). to facilitate removal of the hatching envelope for staining (17). At 55 min PS, eggs were washed 4 times with DMSO in ATA-ASW. For suppression of polar body 1 (FBI) formation, eggs were transferred at 15 min PS to 1 juA/CD in ATA-ASW. At 35 min PS, CD-tr
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 102 P. L. HERTZLER. Figure 2. Effects of cytochalasin D (CD) treatments. For inhibition of polar body 2 (PB2) formation, eggs were transferred at 40 min post-spawning (PS) to 1 /n/W ( mg/1) CD in artificial seawater containing 50 ;uM (ATA-ASW). to facilitate removal of the hatching envelope for staining (17). At 55 min PS, eggs were washed 4 times with DMSO in ATA-ASW. For suppression of polar body 1 (FBI) formation, eggs were transferred at 15 min PS to 1 juA/CD in ATA-ASW. At 35 min PS, CD-treated eggs were washed 4 times with DMSO in ATA-ASW. Control groups ( DMSO carrier control and No Treatment control) were subjected to identical physical manipulations as CD-treated eggs. CD-treated eggs and control groups were fixed with 4% paraformaldehyde in ASW and examined by phase-contrast microscopy. Other samples were fixed in 90% methanol/50 mA/ EGTA. pH and stained with /3-tubulin monoclonal antibody E7 as described previously (14). Nuclei and chromosomes were stained with Sytox green (Molecular Probes, Inc.) for confocal microscopy. (A) DMSO control from 40-55 min PS, dark-field image of embryos at the 16-cell stage, 3-h PS. (B) CD treatment from 40-55 min PS. dark-field image of embryos at 3-h PS. Arrows indicate cleared regions of cytoplasm. (C) DMSO control from 40-55 min PS. embryo at 3-h PS, stained with Sylox green. The image is a composite of optical slices through one-half of the embryo. (D) CD treatment from 40-55 min PS, embryo at 3-h PS. stained with Sytox green. The image is a composite of optical slices through the entire embryo. (E) Phase-contrast image of DMSO control from 15-4(1 min PS, egg at 60 min PS. Note the positions of PBI and PB2 (arrowheads) relative to the hatching envelope. (F) Phase-contrast image of CD treatment from 15-40 min PS, egg at 60 min PS. Two "second" polar bodies (arrowheads) have formed beneath the hatc
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
