Injection of Antisense RNA specific for E-cadherin demonstrates that E-cadherin facilitates compaction, the first differentiative step of the mammalian embryo.
Injection of Antisense RNA specific for E-cadherin demonstrates that E-cadherin facilitates compaction, the first differentiative step of the mammalian embryo.
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注射 E-钙粘蛋白特异性反义 RNA 表明 E-钙粘蛋白促进压缩，这是哺乳动物胚胎分化的第一个步骤。
DOI：
10.1089/ard.1992.2.153
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Erickson,RP
中科院分区：
文献类型：
--
作者：
Ao,A;Erickson,RP
我们研究了反义E-钙粘蛋白RNA在小鼠着床前胚胎中的作用。将反义RNA注射到培养2d的两细胞胚胎的每个细胞内，直到致密正常，并对异常情况进行评分。与注射对照或正义RNA的胚胎相比，注射反义RNA的胚胎表现出延迟的致密化。卵裂延迟不是原因，因为Hoechst染料33258的核染色显示，注射反义RNA的两个未压缩胚胎的核数量与在人绒毛膜促性腺激素(HCG)后相同时间注射正义RNA的致密胚胎的核数量大致相同。在注射反义RNA的胚胎中，E-钙粘蛋白的单抗的免疫荧光比注射的对照胚胎明显减弱，这表明观察到的延迟致密是由于反义RNA抑制了E-钙粘蛋白基因的表达。注射反义RNA的胚胎最终致密，然后表达E-钙粘素，但明显低于对照组。在两细胞阶段注射单个卵裂球会产生半胚胎畸形。
We have investigated the effect of antisense E-cadherin RNA in the preimplantation mouse embryo. Antisense RNA was injected into each cell of two-cell embryos that were cultured for 2 days until the normal time of compaction and scored for abnormalities. Embryos injected with the antisense RNA showed delayed compaction compared to the embryos injected with control, or sense, RNA. Delayed cleavage was not the cause because nuclear staining with Hoechst dye 33258 showed about the same number of nuclei in both uncompacted embryos injected with antisense RNA compared with compacted embryos injected with sense RNA at the same hours post human chorionic gonadotropin (hCG). Immunofluorescence with a monoclonal antibody to E-cadherin was markedly diminished in embryos injected with antisense RNA compared with control, injected embryos, suggesting that the observed delayed compaction is due to the inhibition of E-cadherin gene expression by antisense RNA. Embryos injected with antisense RNA eventually compacted and then expressed E-cadherin, but at lower apparent levels than controls. Injection of a single blastomere at the two-cell stage created half-embryo abnormalities.