Ishida Michiyo, Ohashi Satoshi, Kizaki Yusuke, Naito Jyun-ichiro, Horiguchi Kotaro, Harigaya Toshio
Laboratory of Functional Anatomy, Department of Life Sciences, Faculty of Agriculture, Meiji University, Japan.
J Reprod Dev. 2007 Feb;53(1):69-76. doi: 10.1262/jrd.18002. Epub 2006 Oct 24.
The placenta is a highly differentiated organ essential for embryonic growth and development. In order to search for key molecules that are associated with mouse placental lactogen II (mPL-II) gene expression, we applied mouse cDNA microarray analysis to RNAs extracted from placentae on days 10, 12, 14, 16 and 18 of pregnancy. Changes in gene expression were categorized between days 10 and 12, 12 and 14, 14 and 16 and 16 and 18 of pregnancy. After microarray analysis, which had a minimum detectable fold change for differential expression of 2, we selected 10 genes, Apoa2, Apoc2, Ceacam14, Creg1, Fmo1, Igf2, Slc2a1, Spink3, Spi1-1 and Tpbpa, exhibiting a expression pattern similar to the mPL-II gene. Furthermore, we performed real-time PCR analysis and in situ hybridization (ISH) to find correlative expression genes for the mPL-II gene. From these results, we identified a resemblance in gene expression between mPL-II and Igf2 and selected these genes for performance of double-fluorescence immunohistochemical staining. We colocalized these proteins in labyrinthine trophoblast cells. These results strongly suggest that the expression of mPL-II and Igf2 is highly related to placental development in mice. This large-scale identification of genes regulated during placentogenesis assists in further elucidation of the molecular basis of extraembryonic development and function.
胎盘是胚胎生长发育所必需的高度分化的器官。为了寻找与小鼠胎盘催乳素II(mPL-II)基因表达相关的关键分子,我们将小鼠cDNA微阵列分析应用于从妊娠第10、12、14、16和18天的胎盘提取的RNA。基因表达的变化按妊娠第10和12天、12和14天、14和16天以及16和18天进行分类。在微阵列分析后,其差异表达的最小可检测倍数变化为2,我们选择了10个基因,即载脂蛋白A2(Apoa2)、载脂蛋白C2(Apoc2)、癌胚抗原相关细胞黏附分子14(Ceacam14)、细胞周期相关蛋白1(Creg1)、黄素单加氧酶1(Fmo1)、胰岛素样生长因子2(Igf2)、溶质载体家族2成员1(Slc2a1)、丝氨酸蛋白酶抑制剂Kazal型3(Spink3)、 Spi1-1和滋养层碱性磷酸酶(Tpbpa),它们表现出与mPL-II基因相似的表达模式。此外,我们进行了实时PCR分析和原位杂交(ISH)以寻找mPL-II基因的相关表达基因。从这些结果中,我们发现mPL-II和Igf2在基因表达上存在相似性,并选择这些基因进行双荧光免疫组织化学染色。我们在迷路滋养层细胞中使这些蛋白质共定位。这些结果强烈表明mPL-II和Igf2的表达与小鼠胎盘发育高度相关。胎盘发生过程中调控基因的这种大规模鉴定有助于进一步阐明胚外发育和功能的分子基础。