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目的 构建Csf1rCreRosa26YFP报告基因小鼠，检测Csf1rCre介导黄色荧光素蛋白(YFP)标记单核细胞及不同组织定居巨噬细胞的效率。方法 Rosa26YFP突变小鼠具有一个lox P侧翼的STOP序列，后跟插入Rosa26基因座的黄色荧光蛋白基因(YFP)。当与表达Csf1rCre重组酶的小鼠交配时，STOP序列被删除，并在双突变后代Csf1rCreRosa26YFP的表达组织中观察到YFP表达。将Csf1rCre小鼠与Rosa26YFP小鼠交配，通过PCR筛选出Csf1rCreRosa26YFP报告基因小鼠。分离成年Csf1rCreRosa26YFP小鼠的血液及骨髓单核细胞、肝脏巨噬细胞、肾脏巨噬细胞、肺泡巨噬细胞和脾脏巨噬细胞，标记流式抗体，通过流式细胞术分析Csf1rCre介导YFP标记组织巨噬细胞的效率。结果 Csf1rCreRosa26YFP报告基因小鼠中肾脏、肝脏、脾脏、肺泡巨噬细胞表达YFP中位数为93.25%、92.45%、91.10%、94.70%，血液单核细胞表达YFP中位数为98.20%，骨髓单核细胞表达YFP中位数为93.90%。结论 Csf1rCre可以介导YFP对各组织定居巨噬细胞以及骨髓和血液单核细胞进行示踪。同时，Csf1rCre可用作这些细胞的基因条件性敲除工具鼠。
Abstract：Objective To construct Csf1rCreRosa26YFPmice,and to detect the efficiency of Csf1rCre-mediated yellow fluorescent protein(YFP) labeling monocytes and tissue-resident macrophages in different tissues. Methods The Rosa26YFPmutant mice had a loxP-flanked STOP sequence,followed by a yellow fluorescent protein gene(YFP)inserted into the Rosa26 locus. When crossed with mice expressing the Csf1rCrerecombinase,the STOP sequence was deleted,and yellow fluorescent protein(YFP) expression was observed in the tissues of double-mutant offspring(Csf1rCreRosa26YFP). Csf1rCremice were mated with Rosa26YFPmice,and genotype of Csf1rCreRosa26YFPmice were identified by PCR. Blood and bone marrow monocytes,liver macrophages,kidney macrophages,alveolar macrophages and spleen macrophages of adult Csf1rCreRosa26YFPmice were isolated,labeled with flow cytometry antibodies,and the recombination efficiency of Csf1rCre-mediated YFP labeling tissue macrophages was analyzed by flow cytometry. Results In Csf1rCreRosa26YFPreporter mice,the median percentage of YFP + was 93. 25% in renal macrophages,92. 45% in liver macrophages,and 91. 10% in spleen macrophages. The percentage of YFP +was 94. 70% in alveolar macrophages,98. 20% in blood monocytes,and 93. 90% in bone marrow monocytes.Conclusion Csf1rCrecan be used to trace tissue-resident macrophages as well as bone marrow and blood monocytes. At the same time,Csf1rCrecan target these cells to prepare conditional gene knockout mice.
[1] Mass E,Nimmerjahn F,Kierdorf K,et al. Tissue-specific macrophages:how they develop and choreograph tissue biology[J]. Nat Rev Immunol,2023,23(9):563-79. doi:10. 1038/s41577-023-00848-y.
[2] Pidwill G R,Gibson J F,Cole J,et al. The role of macrophages in Staphylococcus aureus infection[J]. Front Immunol,2021,11:620339. doi:10. 3389/fimmu. 2020. 620339.
[3] Mass E,Ballesteros I,Farlik M,et al. Specification of tissue-resident macrophages during organogenesis[J]. Science,2016,353(6304):aaf4238. doi:10. 1126/science. aaf4238.
[4] Blériot C,Chakarov S,Ginhoux F. Determinants of resident tissue macrophage identity and function[J]. Immunity,2020,52(6):957-70. doi:10. 1016/j. immuni. 2020. 05. 014.
[5] Schulz C,Gomez Perdiguero E,Chorro L,et al. A lineage of myeloid cells independent of Myb and hematopoietic stem cells[J].Science,2012,336(6077):86-90. doi:10. 1126/science.1219179.
[6] Nagata S. Apoptosis and clearance of apoptotic cells[J]. Annu Rev Immunol,2018,36:489-517. doi:10. 1146/annurevimmunol-042617-053010.
[7] Davies L C,Jenkins S J,Allen J E,et al. Tissue-resident macrophages[J]. Nat Immunol,2013,14(10):986-95. doi:10.1038/ni. 2705.
[8] Jenkins S J,Allen J E. The expanding world of tissue-resident macrophages[J]. Eur J Immunol,2021,51(8):1882-96.doi:10. 1002/eji. 202048881.
[9] Shang M,Cappellesso F,Amorim R,et al. Macrophage-derived glutamine boosts satellite cells and muscle regeneration[J]. Nature,2020,587(7835):626-31. doi:10. 1038/s41586-020-2857-9.
[10] Cox N,Crozet L,Holtman I R,et al. Diet-regulated production of PDGFcc by macrophages controls energy storage[J]. Science,2021,373(6550):eabe9383. doi:10. 1126/science. abe9383.
[11] Shi J,Hua L,Harmer D,et al. Cre driver mice targeting macrophages[J]. Methods Mol Biol,2018,1784:263-75. doi:10.1007/978-1-4939-7837-3_24.
[12] Abram C L,Roberge G L,Hu Y,et al. Comparative analysis of the efficiency and specificity of myeloid-Cre deleting strains using Rosa-EYFP reporter mice[J]. J Immunol Methods,2014,408:89-100. doi:10. 1016/j. jim. 2014. 05. 009.
[13] Sahasrabuddhe V,Ghosh H S. Cx3Cr1-Cre induction leads to microglial activation and IFN-1 signaling caused by DNA damage in early postnatal brain[J]. Cell Rep,2022,38(3):110252. doi:10. 1016/j. celrep. 2021. 110252.
[14] Schneider C,Nobs S P,Kurrer M,et al. Induction of the nuclear receptor PPAR-γ by the cytokine GM-CSF is critical for the differentiation of fetal monocytes into alveolar macrophages[J]. Nat Immunol,2014,15(11):1026-37. doi:10. 1038/ni. 3005.
[15] Georgiades P,Ogilvy S,Duval H,et al. Vav Cre transgenic mice:a tool for mutagenesis in hematopoietic and endothelial lineages[J]. Genesis,2002,34(4):251-6. doi:10. 1002/gene.10161.
[16] Hu B,Duan S,Wang Z,et al. Insights into the role of CSF1R in the central nervous system and neurological disorders[J]. Front Aging Neurosci,2021,13:789834. doi:10. 3389/fnagi. 2021.789834.
[17] Chitu V,Richard Stanley E. Regulation of embryonic and postnatal development by the CSF-1 receptor[J]. Curr Top Dev Biol,2017,123:229-75. doi:10. 1016/bs. ctdb. 2016. 10. 004.
[18] Bencheikh L,Diop M K,Rivière J,et al. Dynamic gene regulation by nuclear colony-stimulating factor 1 receptor in human monocytes and macrophages[J]. Nat Commun,2019,10(1):1935.doi:10. 1038/s41467-019-09970-9.
基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.09.001
中图分类号:R392
引用信息:
[1]李晓宇,赵殿元,张胜权,等.Csf1r~(Cre)介导的黄色荧光素蛋白可有效标记各组织定居巨噬细胞[J].安徽医科大学学报,2025,60(09):1567-1573.DOI:10.19405/j.cnki.issn1000-1492.2025.09.001.
基金信息:
国家自然科学基金项目(编号:32270941)~~