ORGANIZATION OF THE SEA-URCHIN EGG ENDOPLASMIC-RETICULUM AND ITS REORGANIZATION AT FERTILIZATION
ORGANIZATION OF THE SEA-URCHIN EGG ENDOPLASMIC-RETICULUM AND ITS REORGANIZATION AT FERTILIZATION
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DOI：
10.1083/jcb.114.5.929
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发表时间：
1991-09-01
影响因子：
7.8
通讯作者：
JAFFE, LA
中科院分区：
文献类型：
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作者：
TERASAKI, M;JAFFE, LA
将荧光二碳菁DiIC18(3)(DiI)的饱和溶液注入大豆油中，对海胆卵的ER进行了染色，染料从油滴扩散到整个卵的ER膜，但不扩散到其他细胞器。共聚焦显微镜显示，巨大的脑池延伸到整个卵子的内部，在皮质处有一个管状膜网络。由于DiI的扩散局限于连续的双层，染料的扩散支持了ER是一个细胞范围的、相互连接的隔室的概念。在时间推移观察中，内池处于连续运动状态，而大脑皮层ER处于静止状态。受精后，内质网似乎变得更加细小，开始出现一个与钙波明显重合的波，并在2-3分钟时最为明显。5-8分钟后，内质网回到与未受精卵相似的组织中。在受精时，皮质网络也发生了变化；它被破坏并最终恢复。DIL标记可以连续观察原核迁移和有丝分裂期间的内质网。DiI染色的膜在迁移到卵子中心时聚集在围绕精子核和中心粒(精子星座)的微管阵列区域；这种聚集在中心体和合子核附近持续整个原核融合和前两个有丝分裂周期。我们使用了一种新的方法来观察活细胞中内质网的时空组织，我们已经证明了受精时内质网的显著重组。
The ER of eggs of the sea urchin Lytechinus pictus was stained by microinjecting a saturated solution of the fluorescent dicarbocyanine DiIC18(3) (DiI) in soybean oil; the dye spread from the oil drop into ER membranes throughout the egg but not into other organelles. Confocal microscopy revealed large cisternae extending throughout the interior of the egg and a tubular membrane network at the cortex. Since diffusion of DiI is confined to continuous bilayers, the spread of the dye supports the concept that the ER is a cell-wide, interconnected compartment. In time lapse observations, the internal cisternae were seen to be in continuous motion, while the cortical ER was stationary. After fertilization, the internal ER appeared to become more finely divided, beginning as a wave apparently coincident with the calcium wave and becoming most marked by 2-3 min. By 5-8 min the ER returned to an organization similar to that of the unfertilized egg. The cortical network also changed at fertilization; it became disrupted and eventually recovered. Dil labeling allowed continuous observations of the ER during pronuclear migration and mitosis. DiI-stained membranes accumulated in the region of the microtubule array surrounding the sperm nucleus and centriole (the sperm aster) as it migrated to the center of the egg; this accumulation persisted near the centrosomes and zygote nucleus throughout pronuclear fusion and the first two mitotic cycles. We have used a new method to observe the spatial and temporal organization of the ER in a living cell, and we have demonstrated a striking reorganization of the ER at fertilization.