Hamaguchi Y, Mabuchi I
Cell Motil. 1982;2(2):103-13. doi: 10.1002/cm.970020203.
Effects of microinjection of phalloidin on fertilization and cleavage of sand dollar (Clypeaster japonicus and Scaphechinus mirabilis) eggs were studied. The drug, previously injected into unfertilized eggs, showed no effect on the elevation of the fertilization membrane upon insemination up to an intracellular concentration of 50 microM. However, the movement of the egg pronucleus to the sperm pronucleus was inhibited and the fusion of pronuclei did not occur. The subsequent development no longer took place. When phalloidin was injected into fertilized eggs, the thickness of the cortical layer increased and the microvilli became conspicuous. Both nuclear division and cleavage were inhibited at the intracellular concentration of more than 20 microM, though the latter seemed to be more sensitive to phalloidin than the former. Fluorescein-labeled phalloidin (FL-phalloidin) was injected into eggs in order to investigate F-actin localization by fluorescence microscopy. In both unfertilized and fertilized eggs, FL-phalloidin was localized in the cortical layer within 1 min after injection. It was also localized in the cortical layer as radially oriented rod-like structures when injected into fertilized eggs before the disappearance of the nuclear membrane. No distinct fluorescence was detected in the mitotic apparatus or in the cleavage furrow. FL-phalloidin redistributed gradually into egg cytoplasm. In unfertilized eggs, fluorescent rods were found especially in the egg pronucleus 30 min after injection.
研究了向海胆（日本饼海胆和奇异光棘球海胆）卵显微注射鬼笔环肽对受精和卵裂的影响。将该药物预先注射到未受精卵中，在细胞内浓度达到50微摩尔时，对授精后受精膜的隆起没有影响。然而，卵原核向精原核的移动受到抑制，原核融合未发生。随后的发育不再进行。当向受精卵中注射鬼笔环肽时，皮质层厚度增加，微绒毛变得明显。在细胞内浓度超过20微摩尔时，核分裂和卵裂均受到抑制，不过后者似乎比前者对鬼笔环肽更敏感。为了通过荧光显微镜研究F-肌动蛋白的定位，将荧光素标记的鬼笔环肽（FL-鬼笔环肽）注射到卵中。在未受精卵和受精卵中，注射后1分钟内FL-鬼笔环肽均定位于皮质层。当在核膜消失前注射到受精卵中时，它也以径向排列的杆状结构定位于皮质层。在有丝分裂器或卵裂沟中未检测到明显的荧光。FL-鬼笔环肽逐渐重新分布到卵细胞质中。在未受精卵中，注射后30分钟，荧光杆尤其出现在卵原核中。
