Hunde, Alemu Regassa: Transcriptome dynamics and molecular cross-talk between bovine oocyte and its companion cumulus cells. - Bonn, 2010. - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn.
Online-Ausgabe in bonndoc: https://nbn-resolving.org/urn:nbn:de:hbz:5N-21866
@phdthesis{handle:20.500.11811/4212,
urn: https://nbn-resolving.org/urn:nbn:de:hbz:5N-21866,
author = {{Alemu Regassa Hunde}},
title = {Transcriptome dynamics and molecular cross-talk between bovine oocyte and its companion cumulus cells},
school = {Rheinische Friedrich-Wilhelms-Universität Bonn},
year = 2010,
month = jul,

volume = 147,
note = {The bi-directional communication between the oocyte and its companion cumulus cells (CCs) is crucial for development and functions of both cell types. The objectives of this study were to identify transcripts that are exclusively expressed either in oocyte or CCs and those which are differentially expressed when the oocyte matures with or with out their companion CCs and vice versa and to investigate functional changes associated with transcripts that are significantly changed during CCs in vitro maturation (IVM). In experiment 1A, CCs were physically removed from oocytes at GV stage by repeated in and out pipetting and the resulting denuded oocytes (DOs) and their companion CCs were frozen. In experiment 1B, intact COCs were cultured; CCs were physically removed and the resulting denuded MII oocytes and their CCs were frozen. In experiment 2, CCs were physically removed from oocytes at GV stage and the resulting (OO-CCs) and other pools of intact oocytes (OO+CCs) were cultured and their CCs were physically removed and both oocytes were frozen. In experiment 3, the ooplasm were micro surgically removed at GV stage and the resulting oocytectomized complexes (CCs-OO) and other intact complexes (CCs+OO) were cultured. The ooplasm were removed from CCs+OO and the resulting MII CCs frozen. In experiment 4, CCs were physically removed from their enclosed oocytes both at GV and MII stages and frozen for subsequent total RNA isolation. In both cases, cells were cultured for 22 hrs and each experiment was repeated three times using pools of biological replicates (n=150). 15 μg of fragmented and biotin labelled cRNA was hybridized with Affymetrix GeneChip®Bovine Genome Array and data were analyzed using linear model for microarray. Significantly changed gene ontology (GO) terms, gene networks and canonical pathways were analyzed using GO consortium and Ingenuity pathway analysis (IPA) respectively. In experiment 1A, of 13162 detected probe sets, 1516 and 2727 are exclusively expressed in GV oocytes and CCs respectively, and 8919 are expressed in both. Similarly, in experiment 1B, of 13602 detected probe sets, 1423 and 3100 are exclusively expressed in MII oocytes and CCs respectively, and 9079 are expressed in both. In experiment 2, 265 transcripts are differentially expressed of which 217 and 48 are over expressed in OO+CCs and OO-CCs, respectively. In experiment 3, of 566 differentially expressed transcripts, 320 and 246 are over expressed in CCs+OO and CCs-OO, respectively. In experiment 4, of 12827 detected probe sets, 4689 and 834 are exclusively expressed in GV and MII CCs respectively, while 7304 are expressed in both. Oocyte specific transcripts include those involved in transcription (IRF6, POU5F1, MYF5, MED18), translation (EIF2AK1, EIF4ENIF1), biopolymer metabolic process (MOS, ACVR1, ZNF529, MAP3K3), DNA replication (MCM6, NASP, ORC6L), protein amino acid phosphorylation (MAP4K2, PRKCH, MOS) and CC specific ones include those involved in macromolecule biosynthetic process (APOA1, USPL1, APOE, NANS), carbohydrate metabolism (HYAL1, PFKL, PYGL, MPI), protein metabolic processes (IHH, APOA1, PLOD1), steroid biosynthetic process (APOA1, CYP11A1, HSD3B1, HSD3B7). 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). Signal transduction, cholesterol biosynthetic processes, DNA replication, cellular growth and proliferation, actin filament polymerisation and cell adhesion are among the top significantly changed biological functions associated with transcripts that are differentially expressed between GV and MII CCs. In conclusion, this study generated large scale gene expression data from different oocyte and CCs samples that would enhance our understanding of the molecular mechanisms underlying oocyte-CCs dialogue in general and oocyte maturation in particular.},
url = {http://hdl.handle.net/20.500.11811/4212}
}

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