Selective expression of eGFP in mouse perivascular astrocytes by modification of the Mlc1 gene using T2A-based ribosome skipping.
Selective expression of eGFP in mouse perivascular astrocytes by modification of the Mlc1 gene using T2A-based ribosome skipping.
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使用基于 T2A 的核糖体跳跃修饰 Mlc1 基因，在小鼠血管周围星形胶质细胞中选择性表达 eGFP。
DOI：
10.1002/dvg.23071
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发表时间：
2017-10
期刊：
影响因子：
--
通讯作者：
McCarty JH
中科院分区：
文献类型：
--
作者：
Toutounchian JJ;McCarty JH
Perivascular astrocyte end feet closely juxtapose cerebral blood vessels to regulate important developmental and physiological processes including endothelial cell proliferation and sprouting as well as the formation of the blood-brain barrier (BBB). The mechanisms underlying these events remain largely unknown due to a lack of experimental models for identifying perivascular astrocytes and distinguishing these cell types from other astroglial populations. Megalencephalic leukoencephalopathy with subcortical cysts 1 (Mlc1) is a transmembrane protein that is expressed in perivascular astrocyte end feet where it controls BBB development and homeostasis. Based on this knowledge, we used T2A peptide-skipping strategies to engineer a knock-in mouse model in which the endogenous Mlc1 gene drives expression of enhanced green fluorescent protein (eGFP), without impacting expression of Mlc1 protein. Analysis of fetal, neonatal and adult Mlc1-eGFP knock-in mice revealed a dynamic spatiotemporal expression pattern of eGFP in glial cells, including nestin-expressing neuroepithelial cells during development and glial fibrillary acidic protein (GFAP)-expressing perivascular astrocytes in the post-natal brain. EGFP was not expressed in neurons, microglia, oligodendroglia, or cerebral vascular cells. Analysis of angiogenesis in the neonatal retina also revealed enriched Mlc1-driven eGFP expression in perivascular astrocytes that contact sprouting blood vessels and regulate blood-retinal barrier permeability. A cortical injury model revealed that Mlc1-eGFP expression is progressively induced in reactive astrocytes that form a glial scar. Hence, Mlc1-eGFP knock-in mice are a new and powerful tool to identify perivascular astrocytes in the brain and retina and characterize how these cell types regulate cerebral blood vessel functions in health and disease.
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影响因子：
25
作者：
John Lin CC;Yu K;Hatcher A;Huang TW;Lee HK;Carlson J;Weston MC;Chen F;Zhang Y;Zhu W;Mohila CA;Ahmed N;Patel AJ;Arenkiel BR;Noebels JL;Creighton CJ;Deneen B
通讯作者：
Deneen B
影响因子：
9.8
作者：
Leegwater, PAJ;Yuan, BQ;van der Knaap, MS
通讯作者：
van der Knaap, MS
影响因子：
3.2
作者：
McCarty JH
通讯作者：
McCarty JH
影响因子：
3.3
作者：
Nagelhus, EA;Mathiisen, TM;Ottersen, OP
通讯作者：
Ottersen, OP
影响因子：
16.2
作者：
Gallo V;Deneen B
通讯作者：
Deneen B