Barhl1 is required for the differentiation of inner ear hair cell-like cells from mouse embryonic stem cells. (March 2018)

• Record Type: Journal Article
• Authors: Zhong, Chao; Chen, Zhenhuang; Luo, Xiaocui; Wang, Cuicui; Jiang, Hui; Shao, Jianzhong; Guan, Minxin; Huang, Liquan; Huang, Xiao; Wang, Jinfu
• Is Part Of: International journal of biochemistry & cell biology. Volume 96(2018)
• Page Start: 79
• Page End: 89
• Publication Date: 2018-03
• Subjects: mESCs mouse embryonic stem cells -- iPSCs induced pluripotent stem cells -- CRISPR clustered regularly interspaced short palindromic repeat -- Cas9 CRISPR- associated protein 9 -- MEF mouse embryonic fibroblast -- EBs embryoid bodies -- EPL early primitive ectoderm-like -- EBMs EPL cell aggregates -- EGF epidermal growth factor -- FGF3 fibroblast growth factor 3 -- FGF10 fibroblast growth factor 10 -- IGF-I insulin-like growth factor-1 -- RA retinoic acid -- FBS fetal bovine serum -- BSA bovine serum albumin -- PBS phosphate buffer saline -- HBSS Hanks' balanced salt solution -- EDTA ethylene diamine tetraacetic acid -- GAPDH glyceraldehyde-3-phosphate dehydrogenase -- DAPI 4′, 6-diamidino-2-phenylindole -- CDS coding sequence -- AP alkaline phosphatase -- qRT-PCR quantitative real-time polymerase chain reaction -- RT-PCR reverse transcription-polymerase chain reaction -- GO gene ontology -- RSAT regulatory sequence analysis tools -- SEM standard error of the mean -- WT wild type; Deafness -- Barhl1 -- CRISPR/Cas9 -- mESCs -- Inner ear hair cells -- Differentiation; Biochemistry -- Periodicals; Cytology -- Periodicals; Biochemistry -- Periodicals; Cell Biology -- Periodicals; Biochimie -- Périodiques; Cytologie -- Périodiques; Biochimie; Cytologie; Biochemistry; Cytology; Ressource Internet (Descripteur de forme); Périodique électronique (Descripteur de forme); Periodicals; 572.05
• DOI: 10.1016/j.biocel.2018.01.013
• Languages: English
• ISSNs: 1357-2725

Abstract

Graphical abstract: Highlights: An efficient in vitro inner ear hair cell induction method is developed. Barhl1 is not involved in the induction of mESCs toward early primitive ectoderm-like (EPL) cells and otic progenitors. Barhl1 is required for the differentiation of hair cell-like cells. Potential Barhl1 direct target genes in hair cells are identified using RNA-sequencing and bioinformatics. Abstract: Inner ear hair cells are mechanoreceptors responsible for hearing. Pathogenic defects of hair cell-specific genes are one of the major causes of deafness. The BarH class homeobox gene Barhl1 is a deafness gene expressed in developing hair cells, yet the role of Barhl1 during hair cell development remains poorly understood. In the present study, we first established an in vitro differentiation system to efficiently obtain mouse embryonic stem cell (mESC)-derived hair cell-like cells. Subsequently, a mESC line carrying a targeted disruption of Barhl1 was generated using CRISPR/Cas9 technology and subjected to the established in vitro hair cell differentiation protocol. Targeted disruption of Barhl1 does not affect the induction of mESCs toward early primitive ectoderm-like (EPL) cells and otic progenitors but strongly inhibits the differentiation of hair cell-like cells. Using RNA-sequencing and bioinformatics, we further unravel the molecular mechanism underlying Barhl1 -mediated hair cell development. Our data demonstrate the essential role of Barhl1 during hair cell development and provide a basis for the treatment of Barhl1 mutation-based deafness.