In vitro attachment of bovine hatched blastocysts on fibronectin is mediated by integrin in a RGD dependent manner.
In vitro attachment of bovine hatched blastocysts on fibronectin is mediated by integrin in a RGD dependent manner.
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DOI：
10.1262/jrd.51.47
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发表时间：
2005-02
期刊：
影响因子：
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通讯作者：
Masashi Takahashi;Misa Takahashi;S. Hamano;Hitomi Takahashi;A. Okano
中科院分区：
文献类型：
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作者：
Masashi Takahashi;Misa Takahashi;S. Hamano;Hitomi Takahashi;A. Okano
我们研究了细胞外基质蛋白对牛孵化囊胚体外附着和生长的影响。体外产生的牛孵化囊胚在纤连蛋白或层粘连蛋白包被的培养皿上培养。孵化的囊胚在纤连蛋白包被的培养皿上粘附并生长，而在层粘连蛋白包被的培养皿上没有观察到附着。含有 Arg-Gly-Asp (RGD) 序列的合成肽存在时，纤连蛋白上的附着和生长受到显着抑制，该序列与纤连蛋白受体（整合素 α5β1）相互作用，但不被含有 Arg-Gly-Glu (RGE) 序列的对照肽抑制。在培养基中添加抗纤连蛋白受体（整合素α5β1）抗体也抑制了纤连蛋白包被的培养皿上的附着和生长。随后，我们分别通过逆转录聚合酶链反应（RT-PCR）和免疫染色检查了孵化囊胚中α5和β1整合素亚基的mRNA表达和蛋白质表达。在囊胚中检测到 mRNA 和蛋白质的表达。这些结果表明，牛孵化囊胚的滋养外胚层细胞已经通过整合素介导的方式获得了体外在纤连蛋白上粘附和生长的能力。
We investigated the effect of extracellular matrix protein on in vitro attachment and outgrowth of bovine hatched blastocysts. In vitro produced bovine hatched blastocysts were cultured on a fibronectin- or laminin-coated Petri dishes. Hatched blastocysts adhered and outgrew on the fibronectin-coated dish whereas no attachment was observed on the laminin-coated dish. The attachment and outgrowth on fibronectin were significantly inhibited in the presence of synthetic peptides containing the Arg-Gly-Asp (RGD) sequence, which interacts with the fibronectin receptor (integrin alpha5beta1), but were not inhibited by the control peptides containing the Arg-Gly-Glu (RGE) sequence. Addition of anti-fibronectin receptor (integrin alpha5beta1) antibody to the culture medium also inhibited the attachment and outgrowth on fibronectin-coated Petri dishes. Subsequently we examined mRNA expression and protein expression of alpha5 and beta1 integrin subunit in the hatched blastocyst by reverse transcription-polymerase chain reaction (RT-PCR) and immunostaining, respectively. Expression of both mRNA and protein were detected in blastocysts. These results indicate that trophectoderm cells of bovine hatched blastocysts have already acquired the ability to adhere and outgrow on fibronectin in vitro by an integrin- mediated manner.