Recent human next-generation sequencing (NGS) studies indicate a correlation between ANKEF1 (ankyrin repeat and EF-hand domain containing protein 1) expression and cilia formation or function. Additionally, a single study conducted in the African clawed frog (Xenopus laevis) showed ankef1 is down-regulated after pharmacological fibroblast growth factor (FGF) inhibition and plays a role in protocadherin-mediated cell protrusion and adhesion. That study also revealed a critical role for ankef1 in the embryonic development of the frog, with morphants exhibiting phenotypes including spina bifida and a shortened body axis. Interestingly, while little is known about ANKEF1 function in other vertebrate systems, recent proteomic data has shown ANKEF1 enriched in ciliated cells. Likewise, publicly available EST profile databases imply ANKEF1 expression in multiple human tissues, including high levels in the testes. Together, these previous studies suggest an important role for ANKEF1 in ciliated tissues and during embryonic development. Here, we report cloning of zebrafish (Danio rerio) ankef1a, as well as its paralog, ankef1b, and expression analyses by whole-mount in situ hybridization (WISH) and quantitative polymerase chain reaction (qPCR) during embryonic development and in adult tissues. WISH shows both forms are ubiquitously expressed early in development, with more discrete expression of both transcripts in embryonic tissues known to precede or possess motile cilia, including dorsal forerunner cells (DFC) and the otic vesicles, respectively. Additionally, both transcripts are enriched in the developing pharynx and swim bladder. Our qPCR results indicate enhanced expression in the testes, along with increased expression in brain. Certainly, our experiments in the zebrafish model system with ankef1a and ankef1b provide a solid foundation for future studies to uncover the molecular pathways through which Ankef1 acts in both healthy and disease states.

最近的人类下一代测序（NGS）研究表明ANKEF1（锚蛋白重复序列​​和包含蛋白质的EF手域）的表达与纤毛形成或功能之间存在相关性。此外，在非洲爪蛙（Xenopus laevis）中进行的一项研究表明，药理性成纤维细胞生长因子（FGF）抑制后ankef1下调，并且在原钙粘蛋白介导的细胞突出和粘附中起作用。该研究还揭示了ankef1的关键作用在青蛙的胚胎发育中，吗啡具有表现出包括脊柱裂和缩短的体轴的表型。有趣的是，虽然对ANKEF1在其他脊椎动物系统中的功能了解甚少，但最近的蛋白质组学数据显示ANKEF1富含纤毛细胞。同样，可公开获得的EST配置文件数据库暗示ANKEF1在多种人体组织中表达，包括睾丸中的高水平表达。总之，这些先前的研究表明ANKEF1在纤毛组织中和胚胎发育过程中具有重要作用。在这里，我们报告了斑马鱼（Danio rerio）ankef1a的克隆及其旁系同源物ankef1b，并在胚胎发育过程中和成年组织中通过全位原位杂交（WISH）和定量聚合酶链反应（qPCR）进行表达分析。WISH显示两种形式都在发育早期广泛表达，两种转录物在已知早于或具有运动性纤毛的胚胎组织中分别离散表达，这些组织分别包括背部前突细胞（DFC）和耳囊。另外，两个转录物均富集在发育中的咽和游泳膀胱中。我们的qPCR结果表明，睾丸中的表达增强，而脑中的表达增加。当然，我们在使用ankef1a和ankef1b的斑马鱼模型系统中进行的实验 为进一步研究揭示Ankef1在健康和疾病状态下发挥作用的分子途径提供了坚实的基础。