Transcriptome analysis reveals an unexpected role of a collagen tyrosine kinase receptor gene, Ddr2, as a regulator of ovarian function.
Transcriptome analysis reveals an unexpected role of a collagen tyrosine kinase receptor gene, Ddr2, as a regulator of ovarian function.
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DOI：
10.1152/physiolgenomics.00073.2009
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发表时间：
2009-10
影响因子：
4.6
通讯作者：
Hirokazu Matsumura;K. Kano;C. Marín de Evsikova;James A. Young;P. Nishina;J. Naggert;K. Naito
中科院分区：
文献类型：
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作者：
Hirokazu Matsumura;K. Kano;C. Marín de Evsikova;James A. Young;P. Nishina;J. Naggert;K. Naito
Slie突变纯合子小鼠缺乏胶原受体，盘状结构域受体2(DDR2)，由于病因不明的外周内分泌系统失调，导致矮小和不育。我们使用系统生物学方法，通过微阵列分析识别受DDR2(Slie/Slie)突变影响的卵巢中的生物网络，并使用分子、细胞和功能生物学分析验证结果。转录组分析表明，在DDR2(Slie/Slie)突变体中，有几个基因类别发生了变化，包括性腺发育、排卵、抗细胞凋亡和类固醇激素。随后的生物学实验证实了转录组分析的预测。例如，DDR2(Slie/Slie)突变体与野生型相比，TUNEL阳性卵泡显著增加，这证实了转录组预测染色质维持减少和抗细胞凋亡。与野生型相比，DDR2(Slie/Slie)突变体的黄体生成素受体和前列腺素E、F受体的表达下降，证实了与排卵有关的激素信号通路。此外，DDR2(Slie/Slie)突变卵巢的体细胞(而不是卵母细胞)中的DDR2和黄体生成素受体的免疫组织化学缺陷表明，生殖细胞存在固有缺陷。事实上，DDR2(Slie/Slie)突变体排卵的卵母细胞明显减少，其卵母细胞能够在体外完成减数分裂和受精。总而言之，我们一致的数据表明，DDR2是卵巢功能中的一个新的关键角色，它作用于体细胞而不是生殖细胞中的经典内分泌途径。
Mice homozygous for the smallie (slie) mutation lack a collagen receptor, discoidin domain receptor 2 (DDR2), and are dwarfed and infertile due to peripheral dysregulation of the endocrine system of unknown etiology. We used a systems biology approach to identify biological networks affected by Ddr2(slie/slie) mutation in ovaries using microarray analysis and validate findings using molecular, cellular, and functional biological assays. Transcriptome analysis indicated several altered gene categories in Ddr2(slie/slie) mutants, including gonadal development, ovulation, antiapoptosis, and steroid hormones. Subsequent biological experiments confirmed the transcriptome analysis predictions. For instance, a significant increase of TUNEL-positive follicles was found in Ddr2(slie/slie) mutants vs. wild type, which confirm the transcriptome prediction for decreased chromatin maintenance and antiapoptosis. Decreases in gene expression were confirmed by RT-PCR and/or qPCR; luteinizing hormone receptor and prostaglandin type E and F receptors in Ddr2(slie/slie) mutants, compared with wild type, confirm hormonal signaling pathways involved in ovulation. Furthermore, deficiencies in immunohistochemistry for DDR2 and luteinizing hormone receptor in the somatic cells, but not the oocytes, of Ddr2(slie/slie) mutant ovaries suggest against an intrinsic defect in germ cells. Indeed, Ddr2(slie/slie) mutants ovulated significantly fewer oocytes; their oocytes were competent to complete meiosis and fertilization in vitro. Taken together, our convergent data signify DDR2 as a novel critical player in ovarian function, which acts upon classical endocrine pathways in somatic, rather than germline, cells.