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. 2019 Jan 29;15(4):757–775. doi: 10.7150/ijbs.28461

Figure 3.

Figure 3

Effect of GPN pretreatment on the CG and vesicles. (A) P. lividus eggs were stained with LysoTracker-RED before and after (40 min) the treatment with 200 µM GPN or DMSO (control), and viewed in confocal microscopy. The changes of the fluorescent signals in the LysoTracker-RED-stained vesicles were examined in the same eggs 25 minutes after fertilization. Control eggs displayed full elevation of the FE, but most eggs treated with GPN failed at the FE elevation despite the formation of multiple fertilization cones (blue arrowheads). (B) Electron micrographs of the cortex of the eggs treated with 200 µM GPN or DMSO. In control eggs, cortical granules (red arrow) positioned underneath the plasma membrane were all exocytosed into the perivitelline space 5 minutes after fertilization (right panel). In GPN-pretreated eggs, CG appeared to fuse with each other (white arrow) or with the adjacent vesicles (black arrow), and to be displaced from the plasma membrane. After insemination, FE elevates only partially while some CG fused with other granules (black arrow) are still visible inside the eggs.