单核细胞过氧化物酶体增殖物激活受体γ条件敲除小鼠的繁殖和基因型鉴定

doi:10.3969/j.issn.1674-5817.2018.03.011

实验动物与比较医学 ›› 2018, Vol. 38 ›› Issue (3): 222-226.DOI: 10.3969/j.issn.1674-5817.2018.03.011

单核细胞过氧化物酶体增殖物激活受体γ条件敲除小鼠的繁殖和基因型鉴定

郑欣洵¹, 张智静¹, 黄宝艺¹, 廖子君², 刘建军³, 罗涛¹

1.北京大学深圳医院,深圳 518035;
2.深圳市第二人民医院,深圳 518035;
3.深圳市疾病预防控制中心,深圳 518000

收稿日期:2017-07-07 修回日期:2018-04-22 出版日期:2018-06-25 发布日期:2021-03-01
作者简介:郑欣洵(1991-),女,广州医科大学麻醉学硕士研究生。E-mail:873444046@qq.com
基金资助:
国家自然科学基金(81271205),深圳市科技创新委员会基础研究(自由探索)项目(201703073000395)

Generation and Genotyping of Monocyte-Peroxisome Proliferator-activated Receptor γ Conditional Knockout Mice

ZHENG Xin-xun¹, ZHANG Zhi-jing¹, HUANG Bao-yi¹, LIAO Zi-jun², LIU Jian-jun³, LUO Tao¹

1. Peking University Shenzhen Hospital,Shenzhen 518035,China;
2. The Second People's Hospital of Shenzhen,Shenzhen 518035,China;
3. Shenzhen Center for Disease Control and Prevention,Shenzhen 518000,China

Received:2017-07-07 Revised:2018-04-22 Online:2018-06-25 Published:2021-03-01

摘要/Abstract

摘要： 目的构建和鉴定单核细胞过氧化物酶体增殖物激活受体γ(PPARγ)条件性基因敲除小鼠。方法将引进的纯合子PPARγ^-loxp小鼠和单核细胞特异性表达cre重组酶的Cx3cr1^cre小鼠进行杂交,得到F1代,用RT-PCR方法鉴定其基因型。将F1代进行自交产生F2代,鉴定基因型,筛选实验所需的PPARγ条件性基因敲除小鼠(实验组, PPARγ-ko)及对照组小鼠(对照组, PPARγ-wt),利用免疫荧光及Western blotting方法鉴定敲除效果。结果筛选出基因型为PPARγ ^loxp/loxp Cx3cr1^cre/-和PPARγ ^loxp/loxp Cx3cr1^cre/cre的小鼠即为本实验所要构建的单核细胞PPARγ条件基因敲除小鼠,基因型为PPARγ ^loxp/loxp Cx3cr1^-/- 小鼠为实验对照组小鼠。结论成功获得单核细胞PPARγ条件性基因敲除小鼠,为单核细胞上PPARγ功能的研究提供小鼠模型。

关键词: 单核细胞过氧化物酶体增殖物激活受体γ(PPARγ), 条件性基因敲除, Cre/loxp系统, 单核细胞

Abstract: Objective To generate monocyte- peroxisome proliferator-activated receptor γ(PPARγ) conditional knockout mice and to identify the genotype.Methods The imported PPARγ-loxp mice and CX3CR1-Cre mice were hybridized to generate F1 generation,and the genotypes were identified by RT-PCR.Using the same method to generate F2 generation and detect their genotypes.Then the effect of PPARγgene knock-out were identified by using immunofluorescence and Western blotting.Result The monocyte-PPARγknockout mice (PPARγ^loxp/loxp Cx3cr1^cre/- or PPARγ^loxp/loxp Cx3cr1^cre/cre) were as test group(PPARγko),and the control group(PPARγwt) of mice (PPARγ^loxp/loxp Cx3cr1^-/-) were also identified.Conclusion Successfully established the monocyte-PPARγknock-out mice,and provide a experimental tool to further study the funtions of PPARγon monocyte .

Key words: Peroxisome proliferator-activated receptor γ(PPARγ), Conditional knockout, Cre-loxp System, Monocyte.

中图分类号:

Q95-33

郑欣洵, 张智静, 黄宝艺, 廖子君, 刘建军, 罗涛. 单核细胞过氧化物酶体增殖物激活受体γ条件敲除小鼠的繁殖和基因型鉴定[J]. 实验动物与比较医学, 2018, 38(3): 222-226.

ZHENG Xin-xun, ZHANG Zhi-jing, HUANG Bao-yi, LIAO Zi-jun, LIU Jian-jun, LUO Tao. Generation and Genotyping of Monocyte-Peroxisome Proliferator-activated Receptor γ Conditional Knockout Mice[J]. Laboratory Animal and Comparative Medicine, 2018, 38(3): 222-226.

参考文献

[1] Wang J,Shen Y,Zhang Y,et al.Recent evidence of the regulatory role of PPARs in neural stem cells and their underlying mechanisms for neuroprotective effects[J].Curr Stem Cell Res Ther,2016,11(3):188-196.
[2] Braissant O,Foufelle F,Scotto C,et al.Differential expression of peroxisome proliferator-activated receptors (PPARs):tissue distribution of PPAR-alpha,-beta,and -gamma in the adult rat[J].Endocrinol,1996,137(1):354-366.
[3] Kapadia R,Yi JH,Vemuganti R,et al.Mechanisms of anti- inflammatory and neuroprotective actions of PPAR-gamma agonists[J].Front Biosci,2008,13:1813-1826.
[4] Odegaard JI,Ricardo-Gonzalez RR,Red Eagle A,et al.Alternative M2 activation of Kupffer cells by PPARdelta ameliorates obesity-induced insulin resistance[J].Cell Metab,2008,7(6):496-507.
[5] Kang K,Reilly SM,Karabacak V,et al.Adipocyte-derived Th2 cytokines and myeloid PPARdelta regulate macrophage polarization and insulin sensitivity[J].Cell Metab,2008,7(6):485-495.
[6] Moraes LA,Piqueras L,Bishop-Bailey D.Peroxisome proliferatoractivated receptors and inflammation[J].Pharmacol Ther,2006,110(3):371-385.
[7] Jiang C,Ting AT,Seed B.PPAR-gamma agonists inhibit production of monocyte inflammatory cytokines[J].Nature,1998,391(6662):82-86.
[8] Mounier R.PPARγtranscription factor controls in anti-inflammatory macrophages the expression of GDF3 that stimulates myogenic cell fusion during skeletal muscle regeneration[J].Med Sci (Paris),2017,33(5):466-469.
[9] Liu N,Zheng JX,Zhuang YS,et al.Anti-Inflammatory Effects of Schisandrin B on LPS-Stimulated BV2 Microglia via Activating PPAR-γ[J].Inflammation,2017,40(3):1006-1011.
[10] 冷潇,周艳,唐海明,等,Lck-Cre×Perpflox/flox条件敲除小鼠的繁育及基因型鉴定[J].成都医学院学报,2014,9(2):114-117.
[11] Rios FJ,Touyz RM,Montezano AC.Isolation and differentiation of murine macrophages[J].Methods Mol Biol,2017,1527:297-309.
[12] Pineda-Torra I,Gage M,de Juan A.Isolation,culture,and polarization of murine bone marrow-derived and peritoneal macrophages[J].Methods Mol Biol,2015,1339:101-109.

单核细胞过氧化物酶体增殖物激活受体γ条件敲除小鼠的繁殖和基因型鉴定

Generation and Genotyping of Monocyte-Peroxisome Proliferator-activated Receptor γ Conditional Knockout Mice

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

本文评价

[1]	杨冠琦, 张君. 芪蓟肾康颗粒总黄酮对大鼠肾小球系膜细胞外基质动态平衡的干预实验[J]. 实验动物与比较医学, 2016, 36(5): 357-360.
[2]	刘宏, 王冰, 黄凯, 项路娟, 孙红梅, 尹燕博. 不同种群比格犬单核细胞、T淋巴细胞检测[J]. 实验动物与比较医学, 2015, 35(6): 495-500.
[3]	陈方明, 潘永明, 陈亮, 施源, 朱科燕, 余旭平, 陈民利. 巴马小型猪动脉粥样硬化模型相关炎症因子的表达[J]. 实验动物与比较医学, 2014, 34(3): 193-198.
[4]	李翔, 史仍飞, 娄淑杰. 玉米肽和有氧运动对肥胖大鼠血浆Glucagon、MCP-1、PP和PYY水平的影响[J]. 实验动物与比较医学, 2014, 34(3): 214-218.
[5]	周炜, 张迪, 于卓, 查巍巍, 冯立新. 精原干细胞中Pten基因敲除小鼠模型的建立及初步的表型分析[J]. 实验动物与比较医学, 2014, 34(1): 1-5.
[6]	董征, 孙雪冬, 满秋红, 姚波, 李玉芳, 刘铁强, 黄珊, 刘广贤, 艾辉胜, 郭梅. nm23-M1基因在小鼠粒单核细胞白血病中的表达[J]. 实验动物与比较医学, 2014, 34(1): 42-45.