Lipid rafts enriched in monosialylGb5Cer carrying the stage-specific embryonic antigen-4 epitope are involved in development of mouse preimplantation embryos at cleavage stage.
Lipid rafts enriched in monosialylGb5Cer carrying the stage-specific embryonic antigen-4 epitope are involved in development of mouse preimplantation embryos at cleavage stage.
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DOI：
10.1186/1471-213x-11-22
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发表时间：
2011-04-14
影响因子：
--
通讯作者：
Kiyokawa N
中科院分区：
文献类型：
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作者：
Sato B;Katagiri YU;Miyado K;Okino N;Ito M;Akutsu H;Okita H;Umezawa A;Fujimoto J;Toshimori K;Kiyokawa N
富含糖鞘糖脂(GSLS)、胆固醇和信号分子的脂筏不仅在配体结合启动的信号转导中发挥重要作用，而且在肌动蛋白的组织、细胞内运输和细胞极性等细胞内事件中也发挥重要作用，但它们在植入前胚胎卵裂分裂中的作用尚不清楚。在这里，我们证明了单唾液酸化Gb5Cer(MSGb5Cer)富含RAFT结构域参与了小鼠植入前胚胎卵裂阶段的发育。在小鼠着床前胚胎中，MSGb5Cer优先定位于卵裂球之间的界面。实时成像分析表明，MSGb5Cer在细胞质分裂过程中定位于卵裂沟中，通过在界面上积累，使其变厚。用甲基-β-环糊精(MbCD)去除细胞膜上的胆固醇，可减少MSGb5Cer的表达并阻止切割。MSGb5Cer与抗MSGb5Cer单抗(6E2)通过交联在界面上大量聚集，导致F-肌动蛋白在界面聚集，并抑制E-钙粘蛋白在界面的定位，导致切割停止。此外，抑制肌动蛋白与细胞松弛素D(CCD)的聚合减少了MSGb5Cer在界面上的积累。在E-钙粘素靶向的胚胎中，MSGb5Cer富含的RAFT膜结构域异位积累。这些结果表明，MSGb5Cer富含的RAFT膜结构域与皮质肌动蛋白网络和E-钙粘蛋白的分布密切相关，参与胞质分裂。
Lipid rafts enriched in glycosphingolipids (GSLs), cholesterol and signaling molecules play an essential role not only for signal transduction started by ligand binding, but for intracellular events such as organization of actin, intracellular traffic and cell polarity, but their functions in cleavage division of preimplantation embryos are not well known. Here we show that monosialylGb5Cer (MSGb5Cer)-enriched raft domains are involved in development during the cleavage stage of mouse preimplantation embryos. MSGb5Cer preferentially localizes at the interfaces between blastomeres in mouse preimplantation embryos. Live-imaging analysis revealed that MSGb5Cer localizes in cleavage furrows during cytokinesis, and that by accumulating at the interfaces, it thickens them. Depletion of cholesterol from the cell membrane with methyl-beta-cyclodextrin (MbCD) reduced the expression of MSGb5Cer and stopped cleavage. Extensive accumulation of MSGb5Cer at the interfaces by cross-linking with anti-MSGb5Cer Mab (6E2) caused F-actin to aggregate at the interfaces and suppressed the localization of E-cadherin at the interfaces, which resulted in the cessation of cleavage. In addition, suppression of actin polymerization with cytochalasin D (CCD) decreased the accumulation of MSGb5Cer at the interfaces. In E-cadherin-targeted embryos, the MSGb5Cer-enriched raft membrane domains accumulated heterotopically. These results indicate that MSGb5Cer-enriched raft membrane domains participate in cytokinesis in a close cooperation with the cortical actin network and the distribution of E-cadherin.