Dlx3b-4b和Gata3是斑马鱼内耳毛细胞发育的主要调节因子

内耳毛细胞是听觉、平衡感觉神经环路的开关，极易损伤、死亡。先天性、获得性毛细胞缺失均导致内耳功能障碍。内耳毛细胞发育受不同信号和转录因子如Fgf， Bmp， Dlx3b-4b和Gata3等调控，但其机理还不十分清楚。敲低Fgf依赖的内耳Dlx3b或Gata3功能导致毛细胞减少，而双敲低导致无毛细胞，说明Dlx3b和Gata3是内耳毛细胞发育调控的核心。本研究计划运用光切割缓释型morpholino技术，诱导型转基因鱼系，CRISPRi/CRISPR-on技术，小分子阻断化合物，TALENs 或CRISPR/Cas9基因组修饰技术、RNA-Seq和ChIP-Seq等手段全面阐明一个基于Fgf，Bmp，Dlx3b-4b和Gata3的斑马鱼内耳毛细胞发育调控网络。本研究有助于加深理解听力、平衡障碍成因，揭示内耳毛细胞命运决定机制和调控新基因，并为内耳毛细胞再生研究提供理论依据。

Mechanosensory hair cells (HCs) of inner ear, delicate, fragile and sensitive to damaging factors, are trigger of hearing or vestibular neural circuitry. Heritable or gained HC defects often lead to otic dysfunctions. Otic HC development is under control of FGF, Bmp, Dlx3b-4b and Gata3 however, the underlying molecular genetic mechanisms are largely unknown. Our preliminary study reveals that Fgf-dependent otic Dlx3b and Gata3, when individually knocked down their functions, results in significant HC loss. Double knock down of both transcription factors completely abolishes HCs, indicating that Dlx3b and Gata3 are center to otic HC development program. We plan to use photo-cleavable morpholino oligos, inducible transgenic lines, CRISPRi/CRISPR-on strategy, small chemical inhibitors, TALENS and CRISPR/Cas9 genome editing tools, RNA-seq and ChIP-seq techniques to unveil a genetic regulation network directing otic HC specification. The network is based on Fgf，Bmp，Dlx3b-4band Gata3, and our proposed study will facilitate our understanding of sensual hearing or vestibular disorders, reveal novel regulators in the genetic network, and provide new insights of HC fate determination and helpful hints for HC regeneration study.