Iruretagoyena J I, Davis W, Bird C, Olsen J, Radue R, Teo Broman A, Kendziorski C, Splinter BonDurant S, Golos T, Bird I, Shah D
MFM Division, OBGYN Department, University of Wisconsin, Madison, WI, USA
Gene Expression Center, University of Wisconsin, Madison, WI, USA.
Mol Hum Reprod. 2014 Jul;20(7):690-700. doi: 10.1093/molehr/gau026. Epub 2014 Mar 27.
The primitive cardiac tube starts beating 6-8 weeks post fertilization in the developing embryo. In order to describe normal cardiac development during late first and early second trimester in human fetuses this study used microarray and pathways analysis and created a corresponding 'normal' database. Fourteen fetal hearts from human fetuses between 10 and 18 weeks of gestational age (GA) were prospectively collected at the time of elective termination of pregnancy. RNA from recovered tissues was used for transcriptome analysis with Affymetrix 1.0 ST microarray chip. From the amassed data we investigated differences in cardiac development within the 10-18 GA period dividing the sample by GA in three groups: 10-12 (H1), 13-15 (H2) and 16-18 (H3) weeks. A fold change of 2 or above adjusted for a false discovery rate of 5% was used as initial cutoff to determine differential gene expression for individual genes. Test for enrichment to identify functional groups was carried out using the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Array analysis correctly identified the cardiac specific genes, and transcripts reported to be differentially expressed were confirmed by qRT-PCR. Single transcript and Ontology analysis showed first trimester heart expression of myosin-related genes to be up-regulated >5-fold compared with second trimester heart. In contrast the second trimester hearts showed further gestation-related increases in many genes involved in energy production and cardiac remodeling. In conclusion, fetal heart development during the first trimester was dominated by heart-specific genes coding for myocardial development and differentiation. During the second trimester, transcripts related to energy generation and cardiomyocyte communication for contractile coordination/proliferation were more dominant. Transcripts related to fatty acid metabolism can be seen as early as 10 weeks and clearly increase as the heart matures. Retinol receptor and gamma-aminobutyric acid (GABA) receptor transcripts were detected, and have not been described previously in human fetal heart during this period. For the first time global gene expression of heart has been described in human samples to create a database of normal development to understand and compare with known abnormal fetal heart development.
原始心管在受精后6 - 8周于发育中的胚胎开始跳动。为了描述人类胎儿妊娠早期末和中期初的正常心脏发育情况,本研究采用了微阵列和通路分析,并创建了一个相应的“正常”数据库。在妊娠终止时前瞻性收集了14例孕龄(GA)在10至18周之间的人类胎儿心脏。从回收组织中提取的RNA用于使用Affymetrix 1.0 ST微阵列芯片进行转录组分析。根据收集的数据,我们将样本按GA分为三组:10 - 12周(H1)、13 - 15周(H2)和16 - 18周(H3),研究了10 - 18 GA期内心脏发育的差异。将2倍或更高的变化倍数调整为5%的错误发现率,作为确定单个基因差异基因表达的初始截断值。使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)进行富集测试以识别功能组。阵列分析正确鉴定了心脏特异性基因,并且通过qRT - PCR证实了报道的差异表达转录本。单转录本和本体分析表明,与妊娠中期心脏相比,妊娠早期心脏中肌球蛋白相关基因的表达上调了5倍以上。相反,妊娠中期心脏显示出许多参与能量产生和心脏重塑的基因随着孕周进一步增加。总之,妊娠早期胎儿心脏发育主要由编码心肌发育和分化的心脏特异性基因主导。在妊娠中期,与能量产生以及心肌细胞通讯以进行收缩协调/增殖相关的转录本更为突出。与脂肪酸代谢相关的转录本早在10周时就可见,并且随着心脏成熟而明显增加。检测到视黄醇受体和γ - 氨基丁酸(GABA)受体转录本,在此期间之前尚未在人类胎儿心脏中描述过。首次在人类样本中描述了心脏的全局基因表达,以创建正常发育数据库,用于理解并与已知的异常胎儿心脏发育进行比较。