Ohkubo Yukihiro, Miyoshi Kazuchika, Yoshida Mitsutoshi
Laboratory of Animal Reproduction, United Graduate School of Agricultural Science, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065, Japan.
Reprod Fertil Dev. 2012;24(2):370-81. doi: 10.1071/RD11048.
The present study was carried out to compare the gene expression profiles of in vitro-generated embryos derived from adult and prepubertal Japanese Black cattle oocytes using GeneChip Bovine Genome Array (containing 24072 probe sets representing over 23000 transcripts). Microarray experiments were performed on populations of 8- to 16-cell stage embryos and blastocysts derived from adult (24-35 months old) versus prepubertal (9-10 months old) Japanese Black cattle oocytes matured and fertilised in vitro. In total, 591 (2.4%) and 490 (2.0%) genes were differentially expressed in prepubertal and adult bovine in 8- to 16-cell and blastocyst stage embryos, respectively. Out of these, 218 and 248 genes were upregulated, while 373 and 242 were downregulated in prepubertal and adult 8- to 16-cell and blastocysts stage embryos, respectively. Gene ontology classification regarding biological process, molecular functions and cellular component revealed diversity in transcript abundances between prepubertal and adult groups in both the distinct developmental stages. Quantitative reverse transcription-PCR validated the expression differences of some selected transcripts as identified by microarray analysis. To our knowledge, this is the first report indicating the significant number of genes differentially expression (>2-fold, P<0.01) in preimplantition embryos between adult and prepubertal Japanese Black cattle during in vitro development.
本研究旨在使用基因芯片牛基因组阵列(包含代表超过23000个转录本的24072个探针组)比较成年和青春期前日本黑牛卵母细胞体外生成胚胎的基因表达谱。对来自成年(24 - 35月龄)和青春期前(9 - 10月龄)日本黑牛卵母细胞体外成熟和受精的8至16细胞期胚胎和囊胚群体进行了微阵列实验。总共,在青春期前和成年牛的8至16细胞期胚胎和囊胚期胚胎中,分别有591个(2.4%)和490个(2.0%)基因差异表达。其中,在青春期前和成年的8至16细胞期胚胎和囊胚期胚胎中,分别有218个和248个基因上调,373个和242个基因下调。关于生物过程、分子功能和细胞成分的基因本体分类显示,在两个不同发育阶段,青春期前和成年组之间的转录本丰度存在差异。定量逆转录 - PCR验证了微阵列分析确定的一些选定转录本的表达差异。据我们所知,这是第一份报告表明在体外发育过程中,成年和青春期前日本黑牛植入前胚胎之间存在大量差异表达基因(>2倍,P<0.01)。