Conditional gene targeting in mouse high endothelial venules.
Conditional gene targeting in mouse high endothelial venules.
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DOI：
10.4049/jimmunol.0802327
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发表时间：
2009-05-01
期刊：
影响因子：
--
通讯作者：
Saga Y
中科院分区：
文献类型：
--
作者：
Kawashima H;Hirakawa J;Tobisawa Y;Fukuda M;Saga Y
高内皮微静脉（HEVs）是次级淋巴器官的特化血管，由具有特征性立方形态的内皮细胞组成。淋巴细胞选择性地粘附并迁移穿过HEV以启动免疫应答。在这项研究中，我们建立了一个新的转基因小鼠系表达Cre重组酶的基因编码戊型肝炎表达的磺基转移酶，N-乙酰葡糖胺-6-O-磺基转移酶2（GlcNAc 6ST-2）的转录控制下，使用细菌人工染色体重组。将这些转基因小鼠与R 0 SA 26报告菌株杂交，R 0 SA 26报告菌株在Cre介导的重组后表达lacZ，并用5-溴-4-氯-5-吲哚基-β-D-半乳糖苷对所得后代进行染色，表明Cre重组酶在次级淋巴器官中的mAb MECA 79反应性HEV中特异性表达，但在转基因小鼠的任何其他血管中均不表达。Cre重组酶的表达与新生儿淋巴结中从不成熟的mAb MECA 367反应性HEV到成熟的mAb MECA 79反应性HEV的发育转换相关。除HEV外，Cre重组酶也在结肠绒毛中强烈表达，其再现了GlcNAc 6ST-2 GFP/GFP敲入小鼠和通过RT-PCR证实的GlcNAc 6ST-2的内在表达。此外，用抗微生物剂处理揭示了Cre重组酶在转基因小鼠中的结肠表达受结肠中的肠道细菌调节。此外，Cre重组酶在脑、睾丸、胃、小肠和肺中的一小部分细胞中表达。鉴于Cre重组酶的限制性表达，该转基因小鼠系应可用于使用Cre/loxP系统阐明组织特异性基因功能。
High endothelial venules (HEVs) are specialized blood vessels of secondary lymphoid organs composed of endothelial cells with a characteristic cuboidal morphology. Lymphocytes selectively adhere to and migrate across HEVs to initiate immune responses. In this study, we established a novel transgenic mouse line expressing Cre recombinase under the transcriptional control of the gene encoding HEV-expressed sulfotransferase, N-acetylglucosamine-6-O-sulfotransferase 2 (GlcNAc6ST-2), using bacterial artificial chromosome recombineering. Crossing these transgenic mice with the ROSA26 reporter strain, which expresses lacZ following Cre-mediated recombination, and staining the resulting progeny with 5-bromo-4-chloro-5-indolyl-β-D-galactoside indicated that Cre recombinase was specifically expressed in mAb MECA79-reactive HEVs in secondary lymphoid organs but not in any other blood vessels of the transgenic mice. The expression of Cre recombinase correlated with a developmental switch, from immature, mAb MECA367-reactive HEVs to mature, mAb MECA79-reactive HEVs in neonatal lymph nodes. In addition to the HEVs, Cre recombinase was also strongly expressed in the colonic villi, which recapitulated the intrinsic expression of GlcNAc6ST-2 as confirmed in GlcNAc6ST-2GFP/GFP knock-in mice and by RT-PCR. Furthermore, treatment with an antimicrobial agent revealed that the colonic expression of Cre recombinase in the transgenic mice was regulated by commensal bacteria in the colon. In addition, Cre recombinase was expressed in a small subset of cells in the brain, testis, stomach, small intestine, and lung. In view of the restricted expression of Cre recombinase, this transgenic mouse line should be useful for elucidating tissue-specific gene functions using the Cre/loxP system.
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影响因子：
7.8
作者：
Bistrup, A;Bhakta, S;Lee, J K;Belov, Y Y;Gunn, M D;Zuo, F R;Huang, C C;Kannagi, R;Rosen, S D;Hemmerich, S
通讯作者：
Hemmerich, S
影响因子：
56.9
作者：
Butcher, EC;Picker, LJ
通讯作者：
Picker, LJ
影响因子：
4.8
作者：
Hiraoka, N;Kawashima, H;Fukuda, M
通讯作者：
Fukuda, M
影响因子：
15.9
作者：
HANNINEN, A;TAYLOR, C;MICHIE, SA
通讯作者：
MICHIE, SA
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL