Ackr3-Venus knock-in mouse lights up brain vasculature.
Ackr3-Venus knock-in mouse lights up brain vasculature.
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DOI：
10.1186/s13041-021-00862-y
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发表时间：
2021-09-28
期刊：
影响因子：
3.6
通讯作者：
Kieffer BL
中科院分区：
文献类型：
--
作者：
Ehrlich AT;Semache M;Couvineau P;Wojcik S;Kobayashi H;Thelen M;Gross F;Hogue M;Le Gouill C;Darcq E;Bouvier M;Kieffer BL
非典型趋化因子受体3，ACKR3，是一种G蛋白偶联受体，它不与G蛋白偶联，而是招募β拦阻蛋白。目前，ACKR3被认为是癌症和心血管疾病的靶点，但人们对ACKR3作为脑部疾病靶点的潜力知之甚少。此外，已经建立了小鼠品系来鉴定表达受体的细胞，但还没有工具来可视化和研究生理条件下的受体本身。在这里，我们设计了一只表达功能性ACKR3-Venus融合蛋白的敲入(KI)小鼠，以直接检测受体，特别是在成人大脑中。在HEK-293细胞中，天然受体和融合受体表现出相似的膜表达、配体诱导的转运和信号特征，表明金星融合不改变受体信号转导。我们还发现，ACKR3-Venus通过共振能量转移实现了对受体贩运的直接实时监测。在ACKR3-Venus敲入小鼠中，我们发现大脑中ACKR3mRNA水平正常，表明基因转录完整。我们完整地定位了14个外周器官和112个脑区的受体表达，发现ACKR3主要定位于这些组织中的血管系统。在外围，受体的分布与以前的报道一致。在脑内，ACKR3在血管内皮细胞、海马GABA能中间神经元和周围神经母细胞中有显著的表达。总之，我们已经培育出具有可追踪的ACKR3受体的Ackr3-Venus敲入小鼠，这将是研究界询问ACKR3生物学和相关疾病的有用工具。网上版载有补充材料，可在10.1186/s13041-021-00862-y查阅。
The atypical chemokine receptor 3, ACKR3, is a G protein-coupled receptor, which does not couple to G proteins but recruits βarrestins. At present, ACKR3 is considered a target for cancer and cardiovascular disorders, but less is known about the potential of ACKR3 as a target for brain disease. Further, mouse lines have been created to identify cells expressing the receptor, but there is no tool to visualize and study the receptor itself under physiological conditions. Here, we engineered a knock-in (KI) mouse expressing a functional ACKR3-Venus fusion protein to directly detect the receptor, particularly in the adult brain. In HEK-293 cells, native and fused receptors showed similar membrane expression, ligand induced trafficking and signaling profiles, indicating that the Venus fusion does not alter receptor signaling. We also found that ACKR3-Venus enables direct real-time monitoring of receptor trafficking using resonance energy transfer. In ACKR3-Venus knock-in mice, we found normal ACKR3 mRNA levels in the brain, suggesting intact gene transcription. We fully mapped receptor expression across 14 peripheral organs and 112 brain areas and found that ACKR3 is primarily localized to the vasculature in these tissues. In the periphery, receptor distribution aligns with previous reports. In the brain there is notable ACKR3 expression in endothelial vascular cells, hippocampal GABAergic interneurons and neuroblast neighboring cells. In conclusion, we have generated Ackr3-Venus knock-in mice with a traceable ACKR3 receptor, which will be a useful tool to the research community for interrogations about ACKR3 biology and related diseases. The online version contains supplementary material available at 10.1186/s13041-021-00862-y.
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DOI：
10.1007/s11481-015-9616-y
发表时间：
2016-03
期刊：
Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology
影响因子：
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作者：
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发表时间：
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期刊：
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影响因子：
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影响因子：
3.1
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影响因子：
3.1
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通讯作者：
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