Function of the small guanosine triphosphate-binding protein RhoA in the process of implantation
Function of the small guanosine triphosphate-binding protein RhoA in the process of implantation
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DOI：
10.1210/jc.85.12.4742
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发表时间：
2000-12-01
影响因子：
5.8
通讯作者：
Yoshimura, Y
中科院分区：
文献类型：
--
作者：
Shiokawa, S;Sakai, K;Yoshimura, Y
小 GTP 酶的 Rho 家族在响应细胞外刺激的细胞运动和形态控制中占据关键地位。 Rho 蛋白触发收缩应力纤维的形成，从而调节细胞运动。我们探讨了 RhoA 在人子宫内膜和蜕膜中的表达和功能。 RhoA 免疫反应性在增殖期和分泌中期主要分布在腺上皮。在蜕膜中，RhoA 的表达在基质细胞和腺上皮中更为明显。通过免疫印迹法评估增殖期和分泌中期子宫内膜以及蜕膜中的RhoA蛋白水平；在所有细胞裂解物中均检测到分子量为 21 kDa 的 RhoA 蛋白单条带。发现培养的人类蜕膜细胞几乎没有肌动蛋白应力纤维。蜕膜细胞在用肉毒梭菌产生的胞外酶 C3 处理后失去了肌动蛋白应力纤维，而用溶血磷脂酸 (LPA) 刺激的人类蜕膜细胞中出现了新的肌动蛋白应力纤维。胚胎从透明带孵化后，小鼠囊胚附着在培养的人类蜕膜细胞上。大多数孵化的囊胚在 24 小时内附着在人类蜕膜细胞上。培养 48 小时后，附着在蜕膜细胞上的囊胚表现出广泛的生长。用 C3 外酶或 LPA 处理蜕膜细胞不会影响囊胚的孵化和附着率，但胚胎在蜕膜细胞上的生长受到 C3 外酶处理的抑制，且呈剂量依赖性。相反，向蜕膜细胞中添加LPA会剂量依赖性地增加蜕膜细胞上胚胎的生长。这些发现表明蜕膜细胞中的 RhoA 对于附着后的胚胎发育和分化很重要。
The Rho family of small GTPases occupies a key position in the control of cell motility and morphology in response to extracellular stimuli. Rho proteins trigger the formation of contractile stress fibers, resulting in regulation of cell motility. We explored the expression and function of RhoA in human endometrium and decidua. RhoA immunoreactivity had a predominantly glandular epithelial distribution in the proliferative phase and midsecretory phase. In decidua, the expression of RhoA was more pronounced in the stromal cells as well as in the glandular epithelium. RhoA protein levels in proliferative phase and midsecretory phase endometrium as well as decidua were evaluated by immunoblotting; a single band of RhoA protein with a molecular mass of 21 kDa was detected in all cell lysates. Cultured human decidual cells were found to have few actin stress fibers. Decidual cells lost their actin stress fibers by the treatment with C3, an exoenzyme produced by Clostridium botulinum, whereas new actin stress fibers appeared in human decidual cells stimulated with lysophosphatidic acid (LPA). Mouse blastocysts became attached to cultured human decidual cells after embryos hatched from the zona pellucida. The majority of hatched blastocysts attached to human decidual cells within 24 h. Blastocysts attached to decidual cells exhibited extensive outgrowth after 48 h in culture. Treatment of decidual cells with C3 exoenzyme or LPA did not affect the rates of hatching and attachment of blastocysts, but outgrowth of embryos on decidual cells was inhibited by C3 exoenzyme treatment in a dose-dependent manner. Contrariwise, addition of LPA to decidual cells dose dependently increased the outgrowth of embryos on decidual cells. These findings suggest that RhoA in decidual cells is important for embryonic development and differentiation after attachment.