Expression of Pou3f3/Brn-1 and Its Genomic Methylation in Developing Auditory Epithelium
Expression of Pou3f3/Brn-1 and Its Genomic Methylation in Developing Auditory Epithelium
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DOI：
10.1002/dneu.20746
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发表时间：
2009-12-01
影响因子：
3
通讯作者：
Matsunaga, Tatsuo
中科院分区：
文献类型：
--
作者：
Mutai, Hideki;Nagashima, Reiko;Matsunaga, Tatsuo
在哺乳动物耳蜗中，感觉细胞（称为毛细胞（HC））和非感觉细胞（如支持细胞（SC）和间充质细胞）通过表达各种功能分子参与适当的听觉功能。在发育过程中，某些基因的表达通过基因组甲基化（主要的表观遗传调节机制之一）受到抑制。我们探讨了基因组区域差异甲基化的大鼠听觉上皮在出生后第1天（P1）和P14使用扩增甲基化位点（AIMS）。将ARMS片段定位于Pou 3f 3/Brn-1的3 '侧翼区。亚硫酸氢盐转化PCR和甲基化特异性定量PCR结果显示，Pou 3f 3及其附近非编码RNA的表达在P14时降低，而AIMS区及其附近CpG岛的甲基化频率在P14时升高。新生DNA甲基转移酶3a和3b的表达也表明了出生后内耳发育过程中表观遗传调控的作用。免疫组化分析显示Pou 3f 3在耳蜗的SC和间充质细胞中特异性表达，并建立了Pou 3f 3是研究内耳发育的新细胞类型标记物。Pou 3f 3或Pou 3f 2加Pou 3f 3缺陷的小鼠在胚胎耳蜗中未表现出任何异常。Pou 3f 3的缺失既不影响HC祖细胞的增殖，也不影响HC祖细胞的分化活性。然而，Pou 3f 3可能对出生后耳蜗的维持或功能发育很重要。这是第一份报告，研究参与的表观遗传调控机制，在发展中的哺乳动物听觉上皮。(C)2009乳清期刊公司开发神经生物学69：913-930，2009
In the mammalian cochlea, both the sensory cells-called hair cells (HCs)-and nonsensory cells such as supporting cells (SCs) and mesenchymal cells participate in proper auditory function through the expression of various functional molecules. During development, expression of certain genes is repressed through genomic methylation, one of the major epigenetic regulatory mechanisms. We explored the genomic regions that were differentially methylated in rat auditory epithelium at postnatal day 1 (P1) and P14 using amplification of intermethylated sites (AIMS). An ARMS fragment was mapped to the 3'-flanking region of Pou3f3/Brn-1. Bisulfite-converted PCR and quantitative methylation-specific PCR showed that the methylation frequency of the AIMS region and the adjacent CpG island was increased at P14, when the expression of Pou3f3 and the noncoding RNAs nearby decreased. Expression of de novo DNA methyltransferases 3a and 3b also suggests a role of epigenetic regulation during postnatal inner ear development. Immunohistochemical analysis showed that Pou3f3 was expressed specifically in the SCs and mesenchymal cells in the cochlea and established that Pou3f3 is a new cell-type marker for studying inner ear development. Mice deficient in Pou3f3 or Pou3f2 plus Pou3f3 did not exhibit any abnormality in the embryonic cochlea. Absence of Pou3f3 affected neither the proliferation nor the differentiation activities of HC progenitor cells. Pou3f3 may, however, be important for the maintenance or functional development of the postnatal cochlea. This is the first report to study involvement of an epigenetic regulatory mechanism in the developing mammalian auditory epithelium. (C) 2009 Whey Periodicals, Inc. Develop Neurobiol 69:913-930, 2009