Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, New York, NY 10029, USA.
Dev Dyn. 2012 Sep;241(9):1385-95. doi: 10.1002/dvdy.23826. Epub 2012 Jul 16.
Xenopus embryonic skin is composed of the superficial layer with defined apicobasal polarity and the inner layer lacking the apical domain. Multiciliated cells (MCCs) originate in the inner layer of the epidermal ectoderm and subsequently migrate to the surface. How MCCs acquire the apicobasal polarity and intercalate into the superficial layer during neurulation is largely unknown. As Rab11-dependent vesicle trafficking has been implicated in ciliary membrane assembly and in apical domain formation in epithelial cells, we assessed the involvement of Rab11 in MCC development.
Here we report that Rab11 is specifically enriched and becomes apically polarized in skin MCCs. Interference with Rab11 function by overexpression of a dominant negative mutant or injection of a specific morpholino oligonucleotide inhibited MCC intercalation into the superficial layer. Dominant negative Rab11-expressing MCC precursors revealed intrinsic apicobasal polarity, characterized by the apical domain, which is not normally observed in inner layer cells. Despite the presence of the apical domain, the cells with inhibited Rab11 function were randomly oriented relative to the plane of the tissue, thereby demonstrating a defect in planar polarity.
These results establish a requirement for Rab11 in MCC development and support a two-step model, in which the initial polarization of MCC precursors is critical for their integration into the superficial cell layer.
非洲爪蟾胚胎皮肤由具有明确的顶底极性的表层和缺乏顶端结构域的内层组成。多纤毛细胞(MCC)起源于表皮外胚层的内层,随后迁移到表面。在神经胚形成过程中,MCC 如何获得顶底极性并插入到浅层仍然知之甚少。由于 Rab11 依赖性囊泡运输与纤毛膜的组装以及上皮细胞中顶端结构域的形成有关,我们评估了 Rab11 在 MCC 发育中的作用。
在这里,我们报告 Rab11 特异性富集并在皮肤 MCC 中呈现顶端极化。通过过表达显性负突变体或注射特定的反义寡核苷酸来干扰 Rab11 功能,抑制了 MCC 插入浅层。表达显性负性 Rab11 的 MCC 前体细胞显示出固有顶底极性,其特征是存在顶端结构域,而内层细胞中通常不存在该结构域。尽管存在顶端结构域,但抑制 Rab11 功能的细胞相对于组织平面随机定向,从而表明平面极性存在缺陷。
这些结果确立了 Rab11 在 MCC 发育中的必要性,并支持了一个两步模型,其中 MCC 前体的初始极化对于它们整合到浅层细胞层至关重要。