华西口腔医学杂志2024，Vol.42Issue(4)：435-443,9.DOI:10.7518/hxkq.2024.2023374
Wnt1-Cre和Pax2-Cre标记的小鼠第一鳃弓颅颌面部神经嵴细胞异质性研究
Heterogeneity of Wnt1-Cre-marked and Pax2-Cre-marked first branchial arch cranial neural crest cells in mice
摘要
Abstract
Objective This study aimed to explore the heterogeneity and gene ontology of Wnt1-Cre-marked and Pax2-Cre-marked first branchial arch cranial neural crest cells(CNCs)in mice.Methods The embryos of Wnt1-Cre;R26RmTmG and Pax2-Cre;R26RmTmG at embryonic day(E)8.0-E9.25 were collected for histological observation.We per-formed immunostaining to compare green fluorescent pro-tein(GFP)-positive CNCs in Pax2-Cre;R26RAi9 and Wnt1-Cre;R26RAi9 mice at E15.5.Single-cell RNA sequencing(scRNA-seq)was used to analyze the first branchial arch GFP-positive CNCs from Wnt1-Cre;R26RmTmG and Pax2-cre;R26RmTmGmice at E10.5.Real time fluorescence quantitative polymerase chain reaction(q-PCR)was performed to val-idate the differential genes.Results Wnt1-Cre-marked and Pax2-Cre-marked CNCs migrated from the neural plateto first and second branchial arches and to the first branchial arch,respectively,at E8.0.Although Wnt1-Cre-marked and Pax2-Cre-marked CNCs were found mostly in cranial-facial tissues,the former had higher expression in palate and tongue.The results of scRNA-seq showed that Pax2-Cre-marked CNCs specifically contributed to osteoblast differentia-tion and ossification,while Wnt1-Cre-marked CNCs participated in limb development,cell migration,and ossification.The q-PCR data also confirmed the results of gene ontology analysis.Conclusion Pax2-Cre mice are perfect experimen-tal animal models for research on first branchial arch CNCs and derivatives in osteoblast differentiation and ossification.关键词
颅颌面部神经嵴细胞/第一鳃弓/Wnt1-Cre/Pax2-Cre/单细胞测序Key words
cranial neural crest cells/first branchial arch/Wnt1-Cre/Pax2-Cre/single-cell RNA sequencing分类
生物科学引用本文复制引用
徐珏,刘双,符宏高,邵美瑛,陈美玲,黄镇..Wnt1-Cre和Pax2-Cre标记的小鼠第一鳃弓颅颌面部神经嵴细胞异质性研究[J].华西口腔医学杂志,2024,42(4):435-443,9.基金项目
National Nature Science Foundation of China(82170918) （82170918）
Natural Science Foundation of Sichuan Prov-ince(2024NSFSC0549) 国家自然科学基金面上项目(82170918) （2024NSFSC0549）
四川省自然科学基金(2024NSFSC0549) （2024NSFSC0549）
