摘要
目的:探讨纤毛内运输(Ift)140基因在小鼠颅缝早闭中的作用及其在成骨细胞中的功能,以揭示Ift140对颅缝发育的影响。方法:通过公共数据库中的小鼠颅缝组织单细胞数据分析,确定表达Ift140基因的细胞类型;构建前成骨细胞条件性Ift140基因敲除小鼠,通过显微-CT(micro-CT)、阿利新蓝-茜素红整体染色、苏木精-伊红染色观察成骨细胞特异性Ift140基因敲除对小鼠颅缝表型的影响;使用免疫荧光染色(ARL13B/Ac-α-tubulin)探究初级纤毛在正常小鼠和成骨细胞特异性Ift140敲除小鼠颅缝中的数量差异;通过荧光双标实验、micro-CT定量分析、牙本质基质蛋白1(DMP1)免疫荧光染色和抗酒石酸酸性磷酸酶(TRAP)染色研究成骨细胞矿化及分化过程中Ift140的作用。结果:单细胞测序分析发现Ift140在成骨相关细胞类群中有较高的表达量,在前成骨细胞条件性敲除Ift140基因后小鼠发生矢状缝颅缝早闭。初级纤毛蛋白免疫荧光染色发现,Ift140基因敲除后小鼠矢状缝中初级纤毛荧光信号不明显,数量减少。Ift140基因缺失导致成骨分化能力减弱,成骨量增加,但矿化质量下降。结论:Ift140通过调控成骨分化和矿化过程影响颅缝发育,导致颅缝早闭。
Objective:To investigate the role of the intraflagellar transport(IFT)140 gene in murine craniosynostosis and its functions in osteoblasts,thereby revealing IFT140's regulatory impact on cranial suture development.Methods:Single-cell data analysis of murine cranial suture tissues from public databases was performed to identify cell types expressing the Ift140 gene.Conditional Ift140 knockout mice in osteoblast progenitors were generated.Cranial suture phenotypes resulting from Ift140 deletion were analyzed using micro-computed tomography(micro-CT),alcian blue-alizarin red whole-mount skeletal staining,and hematoxylin-eosin histology.The role of Ift140 in osteoblast mineralization and differentiation was investigated through dynamic fluorescent double-labeling,quantitative micro-CT analysis,dentin matrix protein 1(DMP1)immunofluorescence staining,and tartrate-resistant acid phosphatase(TRAP)histochemistry.Results:Single-cell sequencing analysis revealed elevated Ift140 expression in osteoblast-lineage cell populations.Conditional Ift140 knockout in osteoblast progenitors induced premature fusion of the sagittal suture in mice.Immunofluorescence staining for primary cilia markers revealed attenuated fluorescence signals and reduced cilium numbers in the sagittal sutures of Ift140 knockout mice compared to controls.Ift140 deficiency impaired osteogenic differentiation capacity,increased bone volume,but compromised mineralization quality.Conclusions:Ift140 affects suture development and causes premature suture closure by regulating the process of osteoblast differentiation and mineralization.
作者
原震林
孙瑶
YUAN Zhenlin;SUN Yao(Shanghai Engineering Research Center of Tooth Restoration and Regeneration&Tongji Research Institute of Stomatology,Shanghai Tongji Stomatological Hospital and Dental School,Tongji University,Shanghai 200072,China)
出处
《口腔生物医学》
2025年第4期189-196,共8页
Oral Biomedicine
基金
国家自然科学基金面上项目(82270963)。
