Li Xiaohui, Zhang Chunyan, Feng Cui, Zhang Zewen, Feng Nannan, Sha Hang, Luo Xiangzhong, Zou Guiwei, Liang Hongwei
Yangtze River Fisheries Research Institude, Chinese Academy of Fishery Sciences, Wuhan 430223, China.
Laboratory of Zooligical Systematics and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding 071002, China.
Animals (Basel). 2024 May 13;14(10):1451. doi: 10.3390/ani14101451.
Bone morphogenetic protein 6 (BMP-6) is a constituent of the TGF-β superfamily, known for its ability to stimulate bone and cartilage formation. The investigation of 's involvement in the formation of intermuscular bones in fish has garnered significant attention in recent years. The rib cage is an important skeletal structure that plays a protective function for internal organs in fish. However, there has been limited research conducted on the effects of the bmp6 gene on rib development. Silver carp is one of four major fish in China, favoured for its affordability and tender muscle. Nevertheless, the presence of numerous intermuscular bones in silver carp significantly hinders the advancement of its palatability and suitability for processing. This study showcases the effective utilisation of CRISPR/Cas9 technology for the purpose of disrupting the gene in silver carp, leading to the creation of chimeras in the P generation, marking the first instance of such an achievement. The chimeras exhibited complete viability, normal appearance, and partial intermuscular bones loss, with approximately 30% of them displaying rib bifurcation or bending. Subsequently, a transcriptome analysis on ribs of P chimeras and wild-type silver carp was conducted, leading to the identification of 934 genes exhibiting differential expression, of which 483 were found to be up-regulated and 451 were found to be down-regulated. The results of the KEGG analysis revealed that the "NF-kappa B signalling pathway", "Hippo signalling pathway", "osteoclast differentiation", and "haematopoietic cell lineage" exhibited enrichment and displayed a significant correlation with bone development. The up-regulated genes such as , , and in pathways may facilitate the proliferation and differentiation of osteoclasts, whereas the down-regulation of genes such as and in pathways may hinder the formation and specialisation of osteoblasts, ultimately resulting in rib abnormalities. This study presents novel findings on the impact of bmp6 gene deletion on the rib development of silver carp, while simultaneously investigating the previously unexplored molecular mechanisms underlying rib defects in fish.
骨形态发生蛋白6（BMP-6）是转化生长因子-β（TGF-β）超家族的一个组成部分，以其刺激骨骼和软骨形成的能力而闻名。近年来，对其在鱼类肌间骨形成中的作用的研究备受关注。肋骨是鱼类体内对内脏起保护作用的重要骨骼结构。然而，关于bmp6基因对肋骨发育影响的研究有限。鲢鱼是中国四大主要鱼类之一，因其价格实惠且肉质鲜嫩而受到青睐。然而，鲢鱼体内大量的肌间骨严重阻碍了其口感的提升和加工适用性。本研究展示了CRISPR/Cas9技术在破坏鲢鱼该基因方面的有效应用，从而在P代产生了嵌合体，这是首次取得这样的成果。这些嵌合体表现出完全的生存能力、正常的外观，并且部分肌间骨缺失，其中约30%显示出肋骨分叉或弯曲。随后，对P代嵌合体和野生型鲢鱼的肋骨进行了转录组分析，鉴定出934个差异表达基因，其中483个上调，451个下调。KEGG分析结果显示，“NF-κB信号通路”“Hippo信号通路”“破骨细胞分化”和“造血细胞谱系”呈现富集，并且与骨骼发育显著相关。通路中的上调基因如 、 和 可能促进破骨细胞的增殖和分化，而通路中 和 等基因的下调可能阻碍成骨细胞的形成和特化，最终导致肋骨异常。本研究揭示了bmp6基因缺失对鲢鱼肋骨发育的影响的新发现，同时研究了鱼类肋骨缺陷背后此前未被探索的分子机制。
