Time-controllable Nkcc1 knockdown replicates reversible hearing loss in postnatal mice.
Time-controllable Nkcc1 knockdown replicates reversible hearing loss in postnatal mice.
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DOI：
10.1038/s41598-017-13997-7
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发表时间：
2017-10-19
影响因子：
4.6
通讯作者：
Tanaka KF
中科院分区：
文献类型：
--
作者：
Watabe T;Xu M;Watanabe M;Nabekura J;Higuchi T;Hori K;Sato MP;Nin F;Hibino H;Ogawa K;Masuda M;Tanaka KF
鉴定特定蛋白质对复发性和部分可逆性听力损失的因果作用一直很困难，因为缺乏提供可逆基因敲除的动物模型。我们已经开发了转基因小鼠品系Actin-tTS：：Nkcc 1 tetO/tetO，用于可操纵地表达耳蜗K+循环蛋白NKCC 1。通过将四环素依赖性转录沉默子与插入Nkcc 1翻译起始位点上游的四环素操纵子序列结合来阻断Nkcc 1转录。四环素衍生物多西环素的管理可逆调节Nkcc 1敲低。从胚胎第0天开始喂食无强力霉素食物的妊娠/哺乳期母亲的后代在出生后第35天（P35）表现出强烈的Nkcc 1表达抑制（约90%下调）和Nkcc 1无效表型。从P0（分娩）开始喂食无强力霉素食物的P35转基因小鼠显示Nkcc 1表达的抑制较弱（约70%下调），听力损失较少，耳蜗结构变化轻微。在P35用多西环素治疗这些小鼠2周，重新激活Nkcc 1转录至对照水平并改善高频听力水平;即，这些多西环素处理的小鼠表现出部分可逆的听力损失。因此，Actin-tTS：：Nkcc 1 tetO/tetO转基因小鼠系的开发提供了通过可逆敲低Nkcc 1研究可变听力损失的小鼠模型。
Identification of the causal effects of specific proteins on recurrent and partially reversible hearing loss has been difficult because of the lack of an animal model that provides reversible gene knockdown. We have developed the transgenic mouse line Actin-tTS::Nkcc1 tetO/tetO for manipulatable expression of the cochlear K+ circulation protein, NKCC1. Nkcc1 transcription was blocked by the binding of a tetracycline-dependent transcriptional silencer to the tetracycline operator sequences inserted upstream of the Nkcc1 translation initiation site. Administration of the tetracycline derivative doxycycline reversibly regulated Nkcc1 knockdown. Progeny from pregnant/lactating mothers fed doxycycline-free chow from embryonic day 0 showed strong suppression of Nkcc1 expression (~90% downregulation) and Nkcc1 null phenotypes at postnatal day 35 (P35). P35 transgenic mice from mothers fed doxycycline-free chow starting at P0 (delivery) showed weaker suppression of Nkcc1 expression (~70% downregulation) and less hearing loss with mild cochlear structural changes. Treatment of these mice at P35 with doxycycline for 2 weeks reactivated Nkcc1 transcription to control levels and improved hearing level at high frequency; i.e., these doxycycline-treated mice exhibited partially reversible hearing loss. Thus, development of the Actin-tTS::Nkcc1 tetO/tetO transgenic mouse line provides a mouse model for the study of variable hearing loss through reversible knockdown of Nkcc1.
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影响因子：
3.3
作者：
Chen J;Zhao HB
通讯作者：
Zhao HB
影响因子：
2.6
作者：
Hoa, Michael;Friedman, Rick A.;Derebery, M. Jennifer
通讯作者：
Derebery, M. Jennifer
DOI：
10.1083/jcb.137.6.1287
发表时间：
1997-06-16
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hasson T;Gillespie PG;Garcia JA;MacDonald RB;Zhao Y;Yee AG;Mooseker MS;Corey DP
通讯作者：
Corey DP
影响因子：
3.2
作者：
Crouch, JJ;Sakaguchi, N;Schulte, BA
通讯作者：
Schulte, BA
影响因子：
5.5
作者：
BOSHER, SK;WARREN, RL
通讯作者：
WARREN, RL