Specific and spatial labeling of P0-Cre versus Wnt1-Cre in cranial neural crest in early mouse embryos.
Specific and spatial labeling of P0-Cre versus Wnt1-Cre in cranial neural crest in early mouse embryos.
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DOI：
10.1002/dvg.23034
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发表时间：
2017-06
期刊：
影响因子：
--
通讯作者：
Liu HX
中科院分区：
文献类型：
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作者：
Chen G;Ishan M;Yang J;Kishigami S;Fukuda T;Scott G;Ray MK;Sun C;Chen SY;Komatsu Y;Mishina Y;Liu HX
P0-Cre和Wnt 1-Cre小鼠系已被广泛用于与loxP-侧翼小鼠组合以标记和遗传修饰神经嵴（NC）细胞及其衍生物。Wnt 1-Cre一直被认为是金标准，并且由于P0-Cre转基因在标记早期胚胎阶段的NC细胞中的时间和空间分布尚不清楚，因此对P0-Cre的特异性存在担忧。我们重新访问了P0-Cre和Wnt 1-Cre模型标记的NC细胞在早期小鼠胚胎，重点是颅NC。我们发现，R26-lacZ Cre报告基因比CAAG-lacZ Cre报告基因在早期胚胎发生过程中更可靠地响应Cre活性。在E8.0-9.5（4-19体节）胚胎的颅侧NC和脊索区检测到P0-Cre中的Cre免疫信号和P0-Cre/R26-lacZ和P0-Cre/R26-RFP胚胎中的报告基因（lacZ和RFP）活性。在迁移的NC细胞中观察到P0-Cre转基因表达，并且在前脑和后脑中更广泛，但在中脑中不明显。P0-Cre和Wnt 1-Cre的Cre分布模式的差异在中脑和后脑区域中是深刻的，即，在Wnt 1-Cre胚胎中脑和P0-Cre胚胎后脑中广泛存在。P0-Cre和Wnt 1-Cre在标记颅NC中的差异可以更好地解释其NC衍生物的差异分布和由Cre驱动的遗传修饰引起的表型。
P0-Cre and Wnt1-Cre mouse lines have been widely used in combination with loxP-flanked mice to label and genetically modify neural crest (NC) cells and their derivatives. Wnt1-Cre has been regarded as the gold standard and there have been concerns about the specificity of P0-Cre because it is not clear about the timing and spatial distribution of the P0-Cre transgene in labeling NC cells at early embryonic stages. We re-visited P0-Cre and Wnt1-Cre models in the labeling of NC cells in early mouse embryos with a focus on cranial NC. We found that R26-lacZ Cre reporter responded to Cre activity more reliably than CAAG-lacZ Cre reporter during early embryogenesis. Cre immunosignals in P0-Cre and reporter (lacZ and RFP) activity in P0-Cre/R26-lacZ and P0-Cre/R26-RFP embryos were detected in the cranial NC and notochord regions in E8.0–9.5 (4–19 somite) embryos. P0-Cre transgene expression was observed in migrating NC cells and was more extensive in the forebrain and hindbrain but not apparent in the midbrain. Differences in the Cre distribution patterns of P0-Cre and Wnt1-Cre were profound in the midbrain and hindbrain regions, i.e., extensive in the midbrain of Wnt1-Cre and in the hindbrain of P0-Cre embryos. The difference between P0-Cre and Wnt1-Cre in labeling cranial NC may provide a better explanation of the differential distributions of their NC derivatives and of the phenotypes caused by Cre-driven genetic modifications.