纤毛转运蛋白140谱系示踪小鼠模型构建

doi:10.3969/j.issn.1005-4979.2021.03.002

《口腔颌面外科杂志》 ›› 2021, Vol. 31 ›› Issue (3): 137-141. doi: 10.3969/j.issn.1005-4979.2021.03.002

纤毛转运蛋白140谱系示踪小鼠模型构建

陈雨贝(), 孙瑶()

上海牙组织修复与再生工程技术研究中心，同济大学口腔医学院，同济大学附属口腔医院口腔种植科，上海 200072

收稿日期:2020-06-17 修回日期:2020-06-27 出版日期:2021-06-28 发布日期:2021-07-08
通讯作者: 孙瑶，教授. E-mail: yaosun@tongji.edu.cn
作者简介:
陈雨贝（1994—），女，江苏苏州人，硕士研究生. E-mail: 564443202@qq.com
基金资助:
国家自然科学基金(81822012)

Generation of ciliary gene IFT140 lineage tracing mouse model

CHEN Yubei(), SUN Yao()

Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Department of Oral Implantology, School and Hospital of Stomatology, Tongji University, Shanghai 200072, China

Received:2020-06-17 Revised:2020-06-27 Online:2021-06-28 Published:2021-07-08

摘要/Abstract

摘要：

目的：研究纤毛转运蛋白140（intraflagellar transport protein 140，IFT140）阳性细胞在小鼠生长发育过程中的动态分布与转归。方法：利用Cre/loxP系统构建IFT140谱系示踪小鼠动物模型，将构建的IFT140-CreER小鼠与R26RtdTomato小鼠进行交配繁殖，获得基因型为IFT140-CreER；R26RtdTomato小鼠。用他莫昔芬（tamoxifen）腹腔注射诱导Cre重组酶表达，并在特定的时间点观察IFT140阳性细胞的分布、分化及转归规律。结果：聚合酶链反应（polymerase chain reaction，PCR）验证小鼠基因型符合IFT140-CreER；R26RtdTomato。结果显示，阳性细胞在纤毛富集的组织中信号清晰，模型构建成功；骨组织中的IFT140阳性细胞首先出现在骨髓中，随着骨骼发育分布于骨小梁及骨皮质表面。结论：本研究成功构建了谱系示踪小鼠模型，为观察IFT140在小鼠体内重要器官尤其是骨组织发育中的作用，提供实验动物模型。

关键词: 细胞谱系示踪, 纤毛转运蛋白, 纤毛转运蛋白140

Abstract:

Objective: To investigate the dynamic distribution and outcome of intraflagellar transport protein 140 (IFT140) positive cells during mouse growth and development. Methods: The IFT140 lineage tracing mouse model was established by Cre/loxP system. The constructed IFT140-CreER mice were mated with R26RtdTomato mice and IFT140-CreER; R26RtdTomato mice were obtained. Cre recombinase was induced by intraperitoneal injection of tamoxifen. The distribution, differentiation and prognosis of IFT140 positive cells were observed at specific time points. Results: The genotype of the IFT140-CreER; R26RtdTomato mice was verified by polymerase chain reaction(PCR). The signal of positive cells in cilia enriched tissues was clear, so the model was constructed successfully. IFT140 positive cells first appeared in bone marrow and distributed on the surface of trabecular and cortical bone during bone development. Conclusion: In this study, a lineage tracing mouse model was successfully constructed to provide an experimental animal model for dynamically observing the role of IFT140 in the development of vital organs in mice, especially bone tissue.

Key words: cell lineage tracing, intraflagellar transport protein, intraflagellar transport protein 140

中图分类号:

R782

陈雨贝, 孙瑶. 纤毛转运蛋白140谱系示踪小鼠模型构建[J]. 《口腔颌面外科杂志》, 2021, 31(3): 137-141.

CHEN Yubei, SUN Yao. Generation of ciliary gene IFT140 lineage tracing mouse model[J]. 《Journal of Oral and Maxillofacial Surgery》, 2021, 31(3): 137-141.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2021/V31/I3/137

图/表 4

图1 IFT140-CreER; R26RtdTomato示踪小鼠模型构建 A. IFT140-CreER小鼠基因敲入位点示意；B. IFT140-CreER; R26RtdTomato示踪小鼠模型构建示意。

Figure 1 Generation of IFT140-CreER; R26RtdTomato mouse model

图2 IFT140-CreER小鼠基因型鉴定 MUT：IFT140-CreER条带；WT：野生型条带。

Figure 2 Genotype identification of IFT140-CreER mice

图3 IFT140阳性细胞在体内的分布 A. IFT140阳性细胞在切牙中的分布（×10）；B. IFT140阳性细胞在磨牙中的分布（×10）；C. IFT140阳性细胞在毛囊中的分布（×20）；D. IFT140阳性细胞在鼻腔中的分布（×4）；E. IFT140阳性细胞在脑组织中的分布（×10）；F. IFT140阳性细胞在舌组织中的分布（×20）。

Figure 3 Distribution of IFT140 positive cells in vivo

图4 在股骨发育早期IFT140阳性细胞的分布 A. tamoxifen注射及收样时间点示意；B、E. P4+3时IFT140阳性细胞在股骨中的分布（×4，×20）；C、F. P4+7时IFT140阳性细胞在股骨中的分布（×4，×20）；D、G. P4+14时IFT140阳性细胞在股骨中的分布（×4，×20）；H. IFT140阳性细胞的面积统计。*表示P<0.05,与对照组（P4+3）比较。

Figure 4 Distribution of IFT140 positive cells in early femur development

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