Cell type-specific transcriptome analysis reveals a major role for Zeb1 and miR-200b in mouse inner ear morphogenesis.
Cell type-specific transcriptome analysis reveals a major role for Zeb1 and miR-200b in mouse inner ear morphogenesis.
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DOI：
10.1371/journal.pgen.1002309
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发表时间：
2011-09
期刊：
影响因子：
4.5
通讯作者：
Strome SE
中科院分区：
文献类型：
--
作者：
Hertzano R;Elkon R;Kurima K;Morrisson A;Chan SL;Sallin M;Biedlingmaier A;Darling DS;Griffith AJ;Eisenman DJ;Strome SE
细胞异质性阻碍了从复杂哺乳动物器官的大规模表达谱中提取功能显著的结果和推断调控网络。哺乳动物内耳由听觉和前庭系统组成，听觉和前庭系统各自由毛细胞、支持细胞、神经元、间充质细胞、其他上皮细胞和血管组成。我们开发了一种新的协议来排序的听觉和前庭组织的新生小鼠内耳到其主要的细胞成分。经分选的细胞的转录组分析鉴定了细胞类型特异性表达簇。计算分析检测了在决定内耳细胞身份方面发挥关键作用的转录因子和microRNA。具体来说，我们的分析揭示了Zeb 1/miR-200 b通路在内耳建立上皮和间充质身份中的作用。此外，我们检测到的ZEB 1通路的调节不当，在内耳的Twirler小鼠，这表现出，除其他表型，畸形的听觉和前庭迷路。ZEB 1/miR-200 b通路的失调与Twirler突变小鼠的听觉和前庭缺陷的相关性揭示了耳聋和平衡障碍的新机制。我们的方法可以用来破译其他听力和平衡小鼠突变体的其他复杂的调控网络。哺乳动物内耳是一个高度复杂的感觉器官，超过100个基因的突变是遗传性人类非综合征性听力损失的基础。然而，人们对耳聋基因下游的信号级联反应知之甚少。全基因组表达谱分析是获得系统水平的生物过程的理解的宝贵工具。我们开发并验证了一个简单而新颖的协议，分离感觉上皮细胞，神经元，血管和间充质的听觉和前庭上皮新生野生型小鼠。我们的方案是基于流式细胞术来分选和捕获标记有针对内源性表达的分化簇（CD）抗原的市售抗体的细胞。使用这种策略，我们确定了Zeb 1和miR 200 b作为小鼠内耳上皮和间充质身份的调节因子，我们进一步确定了Twirler小鼠突变体中Zeb 1突变破坏的信号通路。我们还展示了这种方法用于表征隔室特异性基因和蛋白质-蛋白质网络的实用性。实施这种分离策略来研究具有听力和平衡表型的其他小鼠突变体可以克服理解耳聋基因功能的许多障碍。
Cellular heterogeneity hinders the extraction of functionally significant results and inference of regulatory networks from wide-scale expression profiles of complex mammalian organs. The mammalian inner ear consists of the auditory and vestibular systems that are each composed of hair cells, supporting cells, neurons, mesenchymal cells, other epithelial cells, and blood vessels. We developed a novel protocol to sort auditory and vestibular tissues of newborn mouse inner ears into their major cellular components. Transcriptome profiling of the sorted cells identified cell type–specific expression clusters. Computational analysis detected transcription factors and microRNAs that play key roles in determining cell identity in the inner ear. Specifically, our analysis revealed the role of the Zeb1/miR-200b pathway in establishing epithelial and mesenchymal identity in the inner ear. Furthermore, we detected a misregulation of the ZEB1 pathway in the inner ear of Twirler mice, which manifest, among other phenotypes, malformations of the auditory and vestibular labyrinth. The association of misregulation of the ZEB1/miR-200b pathway with auditory and vestibular defects in the Twirler mutant mice uncovers a novel mechanism underlying deafness and balance disorders. Our approach can be employed to decipher additional complex regulatory networks underlying other hearing and balance mouse mutants. The mammalian inner ear is a highly complex sensory organ, and mutations in more than 100 genes underlie hereditary human non-syndromic hearing loss. Nevertheless, little is known about the signaling cascades downstream of deafness genes. Genome-wide expression profiling is an invaluable tool for gaining systems-level understanding of biological processes. We developed and validated a simple and novel protocol to isolate sensory epithelial cells, neurons, blood vessels, and mesenchyme of auditory and vestibular epithelia from newborn wild-type mice. Our protocol is based on flow cytometry to sort and capture cells labeled with commercially available antibodies to endogenously expressed cluster of differentiation (CD) antigens. Using this strategy, we identified Zeb1 and miR200b as regulators of epithelial and mesenchymal identity in the mouse inner ear, and we further identified the signaling pathway disrupted by the Zeb1 mutation in the Twirler mouse mutant. We also show the utility of this approach for characterizing compartment-specific genes and protein–protein networks. Implementation of this isolation strategy to study other mouse mutants with hearing and balance phenotypes could overcome many of the obstacles to understanding the function of deafness genes.
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DOI：
10.1111/j.1749-6632.2010.05868.x
发表时间：
2010-01-01
期刊：
YEAR IN HUMAN AND MEDICAL GENETICS: NEW TRENDS IN MENDELIAN GENETICS
影响因子：
--
作者：
Raviv, Dorith;Dror, Amiel A.;Avraham, Karen B.
通讯作者：
Avraham, Karen B.
影响因子：
4.3
作者：
Elkon, Ran;Agami, Reuven
通讯作者：
Agami, Reuven
影响因子：
7
作者：
Linhart, Chaim;Halperin, Yonit;Shamir, Ron
通讯作者：
Shamir, Ron
DOI：
10.1073/pnas.94.26.14450
发表时间：
1997-12-23
影响因子：
11.1
作者：
Cohen-Salmon, M;El-Amraoui, A;Petit, C
通讯作者：
Petit, C
影响因子：
30.8
作者：
Lewis, Morag A.;Quint, Elizabeth;Glazier, Anne M.;Fuchs, Helmut;De Angelis, Martin Hrabe;Langford, Cordelia;van Dongen, Stijn;Abreu-Goodger, Cei;Piipari, Matias;Redshaw, Nick;Dalmay, Tamas;Angel Moreno-Pelayo, Miguel;Enright, Anton J.;Steel, Karen P.
通讯作者：
Steel, Karen P.