Department of Life Science, BK21 Plus Program, Graduate School, Chungnam National University, Daejeon 34134, Korea.
School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu 41566, Korea.
Mol Cells. 2017 Dec 31;40(12):945-953. doi: 10.14348/molcells.2017.0216. Epub 2017 Dec 22.
We report the biological functions of a zebrafish homologue of RING-finger protein 152 (rnf152) during embryogenesis. rnf152 was initially identified as a brain-enriched E3 ligase involved in early embryogenesis of zebrafish. Expression of rnf152 was ubiquitous in the brain at 24 hpf but restricted to the eyes, midbrain-hindbrain boundary (MHB), and rhombomeres at 48 hpf. Knockdown of rnf152 in zebrafish embryos caused defects in the eyes, MHB, and rhombomeres (r1-7) at 24 hpf. These defects in rnf152-deficient embryos were analyzed by whole-mount in situ hybridization (WISH) using neuroD, deltaD, notch1a, and notch3 probes. NeuroD expression was abolished in the marginal zone, outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL) of the eyes at 27 hpf. Furthermore, deltaD and notch1a expression was remarkably reduced in the ONL, INL, subpallium, tectum, cerebellum, and rhombomeres (r1-7) at 24 hpf, whereas notch3 expression was reduced in the tectum, cerebellum, and rhombomeres at 24 hpf. Finally, we confirmed that expression of Notch target genes, her4 and ascl1a, also decreased significantly in these areas at 24 hpf. Thus, we propose that Rnf152 is essential for development of the eyes, midbrain and hindbrain, and that Delta-Notch signaling is involved.
我们报告了 RING-finger 蛋白 152（rnf152）在斑马鱼胚胎发生过程中的生物学功能。rnf152 最初被鉴定为一种脑丰富的 E3 连接酶，参与斑马鱼的早期胚胎发生。rnf152 在 24 hpf 时在大脑中普遍表达，但在 48 hpf 时仅限于眼睛、中脑-后脑边界（MHB）和菱脑节（r1-7）。rnf152 在斑马鱼胚胎中的敲低导致 24 hpf 时眼睛、MHB 和菱脑节（r1-7）的缺陷。用神经 D、deltaD、 notch1a 和 notch3 探针进行全胚胎原位杂交（WISH）分析 rnf152 缺陷胚胎中的这些缺陷。在 27 hpf 时，眼睛的边缘区、外核层（ONL）、内核层（INL）和节细胞层（GCL）中的神经 D 表达被消除。此外，在 24 hpf 时，ONL、INL、subpallium、顶盖、小脑和菱脑节（r1-7）中的 deltaD 和 notch1a 表达显著减少，而在 24 hpf 时，顶盖、小脑和菱脑节（r1-7）中的 notch3 表达减少。最后，我们证实这些区域的 Notch 靶基因 her4 和 ascl1a 的表达也在 24 hpf 时显著降低。因此，我们提出 Rnf152 对眼睛、中脑和后脑的发育是必不可少的，并且 Delta-Notch 信号通路参与其中。
