Fate-mapping analysis using Rorb-IRES-Cre reveals apical-to-basal gradient of Rorb expression in mouse cochlea.
Fate-mapping analysis using Rorb-IRES-Cre reveals apical-to-basal gradient of Rorb expression in mouse cochlea.
复制标题
使用 Rorb-IRES-Cre 进行命运图谱分析揭示了小鼠耳蜗中 Rorb 表达的顶端到基底的梯度。
DOI：
10.1002/dvdy.111
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Liu Zhiyong
中科院分区：
文献类型：
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作者：
Li Chao;Wang Yunfeng;Wang Guangqin;Lu Ying;He Shunji;Sun Yuwei;Liu Zhiyong
条件性功能丧失研究广泛使用Cre/loxP系统进行，因为这有助于避免胚胎或新生儿死亡问题。然而，缺乏内耳特异的Cre菌株，因此即使在条件基因敲除研究中也经常发生致死性。结果我们报道了一个Rorb-IRES-Crewkkin小鼠品系，在该品系中，Cre概括了内源性Rorb(RAR相关孤儿受体β)的表达模式。OfRorb‐IRES‐Cre/+；Rosa26‐CAG‐LSL‐tdTomato/+cochlear分析表明，td番茄只有在E12.5时才在顶端转弯表达。Td番茄在E15.5、E18.5和P5的顶回和中回有表达，而在基回的表达最低。然而，在P15和P30，三个轮回的听觉毛细胞(HCs)和支持细胞(SCs)大多为tdTomato+。有趣的是，直到P5才检测到tdTomato+前庭细胞，而在P15和P30出现了一些细胞。最后，我们还证实了RorbmRNA和蛋白在P30的耳蜗生毛细胞和干细胞中的表达。结论我们发现Rorb在耳蜗尖到底的表达呈梯度。耳蜗特异性RorbCre活性和从顶端到基底端的RorbCre活性应该能够区分耳蜗区和前庭区以及耳蜗区的低频和高频区的基因功能。
BackgroundConditional loss‐of‐function studies are widely conducted using the Cre/Loxp system because this helps circumvent embryonic or neonatal lethality problems. However, Cre strains specific to the inner ear are lacking, and thus lethality frequently occurs even in conditional knockout studies.ResultsHere, we report aRorb‐IRES‐Creknockin mouse strain in which the Cre recapitulates the expression pattern of endogenousRorb(RAR‐related orphan receptor beta). Analysis ofRorb‐IRES‐Cre/+;Rosa26‐CAG‐LSL‐tdTomato/+cochlear samples revealed that tdTomato was expressed at the apical turn only by E12.5. TdTomato was observed in the apical and middle turns but was minimally expressed in the basal turn at E15.5, E18.5, and P5. However, most of the auditory hair cells (HCs) and supporting cells (SCs) in all three turns were tdTomato+ at P15 and P30. Intriguingly, no tdTomato+ vestibular cells were detected until P5 and a few cells were present at P15 and P30. Finally, we also confirmedRorbmRNA and protein expression in cochlear HCs and SCs at P30.ConclusionsWe reveal thatRorbexpression exhibits an apical‐to‐basal gradient in cochleae. The cochlear‐specific and apical‐to‐basal‐gradientRorbCre activity should enable discrimination of gene functions in cochlear vs vestibular regions as well as low‐frequency vs high‐frequency regions in the cochlea.