Molecular Control of Patterning in Early Ear Development

早期耳朵发育模式的分子控制

• 批准号: 7854446
• 负责人: KATE Francesca BARALD
• 金额: $ 5.45万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7854446
• 关键词: Adopted; Affect; Afferent Neurons; Alder plant; Animals; Antisense Oligonucleotides; Attention; Biological Models; Cell Count; Cell Differentiation process; Cell Line; Cell Lineage; Cells; Cellular Morphology; Chick Embryo; Cochlea; Complement; Defect; Development; Ear; Embryo; Ganglia; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Goals; Grant; Growth Factor; Hair Cells; Knock-out; Knockout Mice; Label; Laboratories; Labyrinth; Length; Modeling; Molecular; Morphogenesis; Morphology; Mus; Myxoid cyst; Natural regeneration; Neurons; Pattern; Play; Population; Process; Production; RNA Interference; Regulation; Regulator Genes; Role; Sensory; Sensory Hair; Signal Pathway; Staging; Supporting Cell; Testing; Time; Translations; Tretinoin; Undifferentiated; Work; bone morphogenetic protein 4; chordin; gain of function; inhibitor/antagonist; loss of function; model development; mouse model; nerve supply; neuron development; novel; otoconia; overexpression; precursor cell; prevent; promoter; protein expression; research study; transcription factor

DESCRIPTION (provided by applicant): The long-term goals of this work are to understand the molecular signaling pathways involved in the development and regeneration of inner ear sensory and neuronal populations. The studies done during the initial grant period were directed at discovering the upstream genes that regulate expression of the growth factor Bone Morphogenetic Protein 4 (BMP4) and its role in the formation of sensory hair cells (HC) in inner ear development. The work done to date has demonstrated that BMP4/antagonist cascades are critical for normal inner ear morphogenesis (Gerlach et al, 2000;Gerlach-Bank et al, 2002 and Gerlach-Bank et al, 2004) and has identified a novel BMP4 promoter, which is expressed in the inner ear and which is repressed by retinoic acid (RA), explaining why RA-treated inner ears resemble BMP4 antagonist-treated ears (Thompson et al, 2003). In this renewal application, attention is focused on the downstream genes, particularly the role of ZIC genes in inner ear development. The hypothesis that directs this work is that ZIC genes are critical transcription factors that serve as regulatory genes, channeling undifferentiated otocyst precursor cells either to a sensory neuron or sensory hair cell (HC)/supporting cell (SC) fate from a common precursor cell in the otocyst. This laboratory has shown that both ZIC1 and ZIC 2 are expressed at the right time and in the right cells to play critical regulatory roles in this cell lineage selection (Warner et al, 2003). In addition, studies of immortalized inner ear cell lines and ZIC2 knockdown mouse models in this laboratory (Gerlach-Bank et al, in prep.) have provided additional support for this hypothesis. The experiments in this proposal therefore investigate the role of ZIC genes and the atonal class downstream genes they control in sensory cell and sensory neuron lineage specification in the inner ear. For these experiments immortalized otocyst cell lines that represent neuron-like, hair cell- (HC) or supporting cell (SC)-like, and neuronal/sensory precursor-like IMO cell populations (precursor-like) will be used (Germiller et al, submitted). Findings from the cell line experiments will then be tested in functional experiments in "real" inner ears in embryos of chick and ZIC knockout and knockdown mice. The hypotheses to be tested include: ZIC1 genes control sensory neuron formation through regulation of Neurogeninl and NeuroD and ZIC2 genes control sensory hair cell formation in the inner ear by regulating Math1. ZIC1 is also known to downregulate MathI (Ebert et al, 2003) and in some cases in the CNS, BMP4 is thought to upregulate Mathl (Ebert et al, 2003; Alder et al, 1999) but whether this is through effects on ZIC genes is unknown. Gene profiling through microarrays and analyses of loss- and gain-of-function experiments in these model systems will allow tests of all these hypotheses.

描述（由申请人提供）：这项工作的长期目标是了解参与内耳感觉和神经元群体发育和再生的分子信号传导途径。在最初的资助期间进行的研究旨在发现调节生长因子骨形态发生蛋白4（BMP 4）表达的上游基因及其在内耳发育中感觉毛细胞（HC）形成中的作用。迄今为止所做的工作已经证明，BMP 4/拮抗剂级联对于正常的内耳形态发生是至关重要的（盖拉赫等人，2000年; Gerlach-Bank et al，2002和Gerlach-Bank et al，2004），并鉴定了一种新的BMP 4启动子，其在内耳中表达并被视黄酸（RA）抑制，这解释了为什么RA处理的内耳类似于BMP 4拮抗剂处理的耳朵（Thompson等，2003）。在这个更新的申请中，注意力集中在下游基因上，特别是ZIC基因在内耳发育中的作用。指导这项工作的假设是，ZIC基因是作为调控基因的关键转录因子，引导未分化的耳囊前体细胞从耳囊中的共同前体细胞到感觉神经元或感觉毛细胞（HC）/支持细胞（SC）的命运。该实验室已经表明，ZIC 1和ZIC 2都在正确的时间和正确的细胞中表达，从而在该细胞谱系选择中发挥关键的调节作用（Warner et al，2003）。此外，在该实验室中永生化内耳细胞系和ZIC 2敲低小鼠模型的研究（Gerlach-Bank等，准备中）。为这一假设提供了额外的支持。因此，本提案中的实验研究了ZIC基因及其控制的无调类下游基因在内耳感觉细胞和感觉神经元谱系特化中的作用。对于这些实验，将使用代表神经元样、毛细胞（HC）或支持细胞（SC）样和神经元/感觉神经元样IMO细胞群（神经元样）的永生化耳囊细胞系（Germiller等人，提交）。然后，将在鸡胚胎和ZIC敲除和敲除小鼠的“真实的”内耳的功能实验中测试来自细胞系实验的发现。待检验的假设包括：ZIC 1基因通过调节Neurogeninl和NeuroD来控制感觉神经元形成，ZIC 2基因通过调节Math 1来控制内耳中感觉毛细胞形成。还已知ZIC 1下调MathI（Ebert et al，2003），并且在CNS中的某些情况下，认为BMP 4上调MathI（Ebert et al，2003;桤木et al，1999），但这是否是通过对ZIC基因的影响尚不清楚。通过微阵列的基因分析和在这些模型系统中的功能丧失和获得实验的分析将允许测试所有这些假设。

项目成果

A drop array culture for patterning adherent mouse embryonic stem cell-derived neurospheres.

用于对贴壁小鼠胚胎干细胞衍生的神经球进行图案化的液滴阵列培养物。

• DOI: 10.1002/term.2389
• 发表时间: 2018
• 期刊: Journal of tissue engineering and regenerative medicine
• 影响因子: 3.3
• 作者: Dixon,AngelaR;Ramirez,Yadah;Haengel,Kathryn;Barald,KateF
• 通讯作者: Barald,KateF

A student team in a University of Michigan biomedical engineering design course constructs a microfluidic bioreactor for studies of zebrafish development.

• DOI: 10.1089/zeb.2008.0572
• 发表时间: 2009-06
• 期刊: Zebrafish
• 影响因子: 2
• 作者: Shen YC;Li D;Al-Shoaibi A;Bersano-Begey T;Chen H;Ali S;Flak B;Perrin C;Winslow M;Shah H;Ramamurthy P;Schmedlen RH;Takayama S;Barald KF
• 通讯作者: Barald KF

Spatiotemporal expression of Zic genes during vertebrate inner ear development.

• DOI: 10.1002/dvdy.23978
• 发表时间: 2013-07
• 期刊: DEVELOPMENTAL DYNAMICS
• 影响因子: 2.5
• 作者: Chervenak, Andrew P.;Hakim, Ibrahim S.;Barald, Kate F.
• 通讯作者: Barald, Kate F.

Cadherin-11 controls otolith assembly: evidence for extracellular cadherin activity.

• DOI: 10.1002/dvdy.22015
• 发表时间: 2009-08
• 期刊: DEVELOPMENTAL DYNAMICS
• 影响因子: 2.5
• 作者: Clendenon, Sherry G.;Shah, Bijal;Miller, Caroline A.;Schmeisser, Glen;Walter, Amanda;Gattone, Vincent H., II;Barald, Kate F.;Liu, Qin;Marrs, James A.
• 通讯作者: Marrs, James A.

The role of Zic genes in inner ear development in the mouse: Exploring mutant mouse phenotypes.

• DOI: 10.1002/dvdy.24186
• 发表时间: 2014-11
• 期刊: Developmental dynamics : an official publication of the American Association of Anatomists
• 作者: Chervenak AP;Bank LM;Thomsen N;Glanville-Jones HC;Jonathan S;Millen KJ;Arkell RM;Barald KF
• 通讯作者: Barald KF