Loxhd1b inhibits the hair cell development in zebrafish: Possible relation to the BDNF/TrkB/ERK pathway.
Loxhd1b inhibits the hair cell development in zebrafish: Possible relation to the BDNF/TrkB/ERK pathway.
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DOI：
10.3389/fncel.2022.1065309
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发表时间：
2022
影响因子：
5.3
通讯作者：
Wang, Dawei
中科院分区：
文献类型：
--
作者：
Liu, Jingwen;Zhang, Xu;Zhang, Qingchen;Wang, Rongrong;Ma, Jingyu;Bai, Xiaohui;Wang, Dawei
脂氧合酶同源结构域1（LOXHD 1）突变导致常染色体隐性遗传，导致患者高频和中频听力损失。迄今为止，关于LOXHD 1基因表达定位的研究有限。本研究旨在观察Loxhd 1b在斑马鱼、C57 BL/6小鼠耳蜗和HEI-OC 1细胞中的表达。采用斑马鱼胚胎原位杂交技术，研究Loxhd 1b基因在斑马鱼听觉系统中的表达。免疫荧光染色分析Loxhd 1b在C57 BL/6小鼠耳蜗和HEI-OC 1细胞中的表达。采用共聚焦显微活体成像技术，检测敲除Loxhd 1b基因的斑马鱼侧线神经瘤和内耳毛细胞的数量和形态特征。利用显微镜观察敲低Loxhd 1b基因对斑马鱼耳石和半规管发育的影响。转录组测序用于鉴定下游分子和相关信号通路，并通过蛋白质印迹、免疫染色和拯救实验进行验证。原位杂交结果显示，Loxhd 1b在斑马鱼内耳和嗅孔中均有表达，免疫组化结果显示，Loxhd 1在C57 BL/6小鼠耳蜗和HEI-OC 1细胞中均有表达。loxhd 1b基因敲低导致斑马鱼内耳脊髓和侧线神经瘤数量减少，伴随听力功能减弱，还导致耳石和耳滤泡发育缺陷。转录组学分析结果揭示了下游分子脑源性神经营养因子（BDNF），并基于Western印迹、免疫染色和拯救实验验证了Loxhd 1b和BDNF通过协同调节BDNF/TrkB/ERK通路来调节斑马鱼毛细胞的形成。这是第一次发现BDNF/TrkB/ERK通路在Loxhd 1b对斑马鱼毛细胞发育和听觉发育的分子调控中起关键作用。
Mutations in lipoxygenase homology domain 1 (LOXHD1) cause autosomal recessive inheritance, leading to high-frequency and intermediate-frequency hearing losses in patients. To date, studies on the localization of LOXHD1 gene expression are limited. In this study, we aimed to observe the expressions of Loxhd1b in zebrafish, C57BL/6 murine cochlea, and HEI-OC1 cells. The expression of Loxhd1b in the auditory system of zebrafish was explored by in situ hybridization experiments of zebrafish embryos. The expression of Loxhd1b in cochlear and HEI-OC1 cells of C57BL/6 mice was analyzed by immunofluorescence staining. Confocal microscopic in vivo imaging was used to detect the number and morphological characteristics of lateral line neuromasts and inner ear hair cells in zebrafish that knocked down Loxhd1b gene. The effect of knockdown Loxhd1b gene on the development of zebrafish otolith and semicircular canal was observed using microscopic. Transcriptome sequencing was used to identify downstream molecules and associated signaling pathways and validated by western blotting, immunostaining, and rescue experiments. Results of the in situ hybridization with zebrafish embryos at different time points showed that Loxhd1b was expressed in zebrafish at the inner ear and olfactory pores, while the immunostaining showed that Loxhd1 was expressed in both C57BL/6 mouse cochlea and HEI-OC1 cells. Loxhd1b knockdown causes a decrease in the number of spinal and lateral line neuromasts in the inner ear of zebrafish, accompanied by weakened hearing function, and also leads to developmental defects of otoliths and ear follicles. The results of transcriptomics analysis revealed the downstream molecule brain-derived neurotrophic factor (BDNF) and verified that Loxhd1b and BDNF regulate the formation of zebrafish hair cells by synergistic regulation of BDNF/TrkB/ERK pathway based on western blotting, immunostaining, and rescue experiments. This was the first time that the BDNF/TrkB/ERK pathway was identified to play a critical role in the molecular regulation of the development of zebrafish hair cells and the auditory development by Loxhd1b.
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影响因子：
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作者：
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影响因子：
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作者：
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影响因子：
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作者：
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DOI：
10.1007/978-3-319-21059-9_18
发表时间：
2016-01-01
期刊：
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影响因子：
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作者：
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通讯作者：
Ramcharitar, John