Institute of Animal Science, Animal Breeding and Husbandry Group, University of Bonn, Germany.
BMC Genomics. 2011 Jan 24;12:57. doi: 10.1186/1471-2164-12-57.
The bi-directional communication between the oocyte and its companion cumulus cells (CCs) is crucial for development and functions of both cell types. Transcripts that are exclusively expressed either in oocytes or CCs and molecular mechanisms affected due to removal of the communication axis between the two cell types is not investigated at a larger scale. The main objectives of this study were: 1. To identify transcripts exclusively expressed either in oocyte or CCs and 2. To identify those which are differentially expressed when the oocyte is cultured with or without its companion CCs and vice versa.
We analyzed transcriptome profile of different oocyte and CC samples using Affymetrix GeneChip Bovine Genome array containing 23000 transcripts. Out of 13162 genes detected in germinal vesicle (GV) oocytes and their companion CCs, 1516 and 2727 are exclusively expressed in oocytes and CCs, respectively, while 8919 are expressed in both. Similarly, of 13602 genes detected in metaphase II (MII) oocytes and CCs, 1423 and 3100 are exclusively expressed in oocytes and CCs, respectively, while 9079 are expressed in both. A total of 265 transcripts are differentially expressed between oocytes cultured with (OO+CCs) and without (OO-CCs) CCs, of which 217 and 48 are over expressed in the former and the later groups, respectively. Similarly, 566 transcripts are differentially expressed when CCs mature with (CCs+OO) or without (CCs-OO) their enclosed oocytes. Of these, 320 and 246 are over expressed in CCs+OO and CCs-OO, respectively.While oocyte specific transcripts include those involved in transcription (IRF6, POU5F1, MYF5, MED18), translation (EIF2AK1, EIF4ENIF1) and CCs specific ones include those involved in carbohydrate metabolism (HYAL1, PFKL, PYGL, MPI), protein metabolic processes (IHH, APOA1, PLOD1), steroid biosynthetic process (APOA1, CYP11A1, HSD3B1, HSD3B7). Similarly, while transcripts over expressed in OO+CCs are involved in carbohydrate metabolism (ACO1, 2), molecular transport (GAPDH, GFPT1) and nucleic acid metabolism (CBS, NOS2), those over expressed in CCs+ OO are involved in cellular growth and proliferation (FOS, GADD45A), cell cycle (HAS2, VEGFA), cellular development (AMD1, AURKA, DPP4) and gene expression (FOSB, TGFB2).
In conclusion, this study has generated large scale gene expression data from different oocyte and CCs samples that would provide insights into gene functions and interactions within and across different pathways that are involved in the maturation of bovine oocytes. Moreover, the presence or absence of oocyte and CC factors during bovine oocyte maturation can have a profound effect on transcript abundance of each cell types, thereby showing the prevailing molecular cross-talk between oocytes and their corresponding CCs.
卵母细胞与其伴生的卵丘细胞(CCs)之间的双向通讯对于两种细胞类型的发育和功能都至关重要。尚未在更大范围内研究仅在卵母细胞或 CCs 中特异性表达的转录本,以及由于两种细胞类型之间通讯轴的去除而受到影响的分子机制。本研究的主要目的是:1. 鉴定仅在卵母细胞或 CCs 中特异性表达的转录本,2. 鉴定当卵母细胞在有或没有其伴生 CCs 培养时以及反之亦然时差异表达的那些转录本。
我们使用包含 23000 个转录本的 Affymetrix GeneChip Bovine Genome 阵列分析了不同卵母细胞和 CC 样本的转录组谱。在处于生发泡(GV)期的卵母细胞及其伴生 CCs 中检测到的 13162 个基因中,分别有 1516 和 2727 个基因仅在卵母细胞和 CCs 中特异性表达,而 8919 个基因在两者中均有表达。同样,在处于 MII 期的卵母细胞和 CCs 中检测到的 13602 个基因中,分别有 1423 和 3100 个基因仅在卵母细胞和 CCs 中特异性表达,而 9079 个基因在两者中均有表达。在有(OO+CCs)和没有(OO-CCs)CCs 培养的卵母细胞之间有 265 个转录本存在差异表达,其中 217 和 48 个在前者和后者中表达上调。同样,当 CCs 在有(CCs+OO)或没有(CCs-OO)其内含卵母细胞时成熟时,有 566 个转录本存在差异表达。其中,在 CCs+OO 和 CCs-OO 中分别有 320 和 246 个转录本表达上调。卵母细胞特异性转录本包括参与转录(IRF6、POU5F1、MYF5、MED18)、翻译(EIF2AK1、EIF4ENIF1)的转录本,CCs 特异性转录本包括参与碳水化合物代谢(HYAL1、PFKL、PYGL、MPI)、蛋白质代谢过程(IHH、APOA1、PLOD1)、类固醇生物合成过程(APOA1、CYP11A1、HSD3B1、HSD3B7)的转录本。同样,在 OO+CCs 中表达上调的转录本参与碳水化合物代谢(ACO1、2)、分子运输(GAPDH、GFPT1)和核酸代谢(CBS、NOS2),在 CCs+OO 中表达上调的转录本参与细胞生长和增殖(FOS、GADD45A)、细胞周期(HAS2、VEGFA)、细胞发育(AMD1、AURKA、DPP4)和基因表达(FOSB、TGFB2)。
总之,本研究从不同的卵母细胞和 CCs 样本中生成了大规模的基因表达数据,这将为研究参与牛卵母细胞成熟的不同途径内和途径之间的基因功能和相互作用提供深入的了解。此外,牛卵母细胞成熟过程中卵母细胞和 CCs 因子的存在或缺失会对每个细胞类型的转录本丰度产生深远影响,从而显示出卵母细胞与其相应 CCs 之间普遍存在的分子串扰。