vwa1 Knockout in Zebrafish Causes Abnormal Craniofacial Chondrogenesis by Regulating FGF Pathway.
vwa1 Knockout in Zebrafish Causes Abnormal Craniofacial Chondrogenesis by Regulating FGF Pathway.
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DOI：
10.3390/genes14040838
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发表时间：
2023-03-30
期刊：
影响因子：
3.5
通讯作者：
Chen, Xiaowei
中科院分区：
文献类型：
--
作者：
Niu, Xiaomin;Zhang, Fuyu;Ping, Lu;Wang, Yibei;Zhang, Bo;Wang, Jian;Chen, Xiaowei
半面短小症 (HFM) 是一种罕见的第一和第二咽弓发育障碍，与 VWA1（包含 1 的冯维勒布兰德因子 A 结构域）的点突变有关，该突变在五代谱系中编码蛋白质 WARP。然而，VWA1 突变与 HFM 发病机制的关系尚不清楚。在这里，我们试图通过使用 CRISPR/Cas9 生成 vwa1 敲除斑马鱼品系，在分子水平上阐明 VWA1 突变的影响。突变体和脆性体表现出软骨畸形，包括发育不全的梅克尔软骨和腭方软骨、畸形的角软骨、角鳃软骨畸形或缺失。软骨细胞表现出较小的尺寸和纵横比，并且排列不规则。原位杂交和RT-qPCR显示barx1和col2a1a表达减少，表明颅神经嵴细胞（CNCC）凝集和分化异常。突变体中 CNCC 的增殖和存活也受到损害。 FGF 通路组分（包括 fgf8a、fgfr1、fgfr2、fgfr3、fgfr4 和 runx2a）的表达下降，这表明 VWA1 在调节 FGF 信号传导中发挥作用。我们的结果表明，VWA1 通过影响 CNCC 的浓缩、分化、增殖和凋亡而对斑马鱼软骨形成至关重要，并且可能通过调节 FGF 途径影响软骨形成。
Hemifacial microsomia (HFM), a rare disorder of first- and second-pharyngeal arch development, has been linked to a point mutation in VWA1 (von Willebrand factor A domain containing 1), encoding the protein WARP in a five-generation pedigree. However, how the VWA1 mutation relates to the pathogenesis of HFM is largely unknown. Here, we sought to elucidate the effects of the VWA1 mutation at the molecular level by generating a vwa1-knockout zebrafish line using CRISPR/Cas9. Mutants and crispants showed cartilage dysmorphologies, including hypoplastic Meckel’s cartilage and palatoquadrate cartilage, malformed ceratohyal with widened angle, and deformed or absent ceratobranchial cartilages. Chondrocytes exhibited a smaller size and aspect ratio and were aligned irregularly. In situ hybridization and RT-qPCR showed a decrease in barx1 and col2a1a expression, indicating abnormal cranial neural crest cell (CNCC) condensation and differentiation. CNCC proliferation and survival were also impaired in the mutants. Expression of FGF pathway components, including fgf8a, fgfr1, fgfr2, fgfr3, fgfr4, and runx2a, was decreased, implying a role for VWA1 in regulating FGF signaling. Our results demonstrate that VWA1 is essential for zebrafish chondrogenesis through effects on condensation, differentiation, proliferation, and apoptosis of CNCCs, and likely impacts chondrogenesis through regulation of the FGF pathway.
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DOI：
10.1038/nrm3528
发表时间：
2013-03
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
--
作者：
Mork L;Crump G
通讯作者：
Crump G
影响因子：
3.6
作者：
Grabb, W C
通讯作者：
Grabb, W C
影响因子：
4.8
作者：
Allen, Justin M.;Zamurs, Laura;Bateman, John F.
通讯作者：
Bateman, John F.
影响因子：
2.7
作者：
Lee, KH;Xu, QH;Breitbart, RE
通讯作者：
Breitbart, RE