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Cited by 26Pluripotent Stem Cells ResearchDevelopmental Biology and Gene RegulationAnimal Genetics and Reproduction

A mRNA Landscape of Bovine Embryos after Standard and MAPK-Inhibited Culture Conditions: a Comparative Analysis

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Brinkhof, Bas, van Tol, Helena TA, Groot Koerkamp, Marian JA, Riemers, Frank M, IJzer, Sascha G, Mashayekhi, Kaveh, Haagsman, Henk P, Roelen, Bernard AJ·2015·BMC Genomics
DOI 10.1186/s12864-015-1448-xPMC PMC4397860
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Abstract

BACKGROUND: Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed. RESULTS: Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated. CONCLUSION: The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells.

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    “However, a number of studies used different culture temperatures (Brinkhof et al., 2015; Xu et al., 2017), which highlights the need to declare and monitor the incubation temperature.”

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Authors
Brinkhof, B., van Tol, H. T., Groot Koerkamp, M. J., Riemers, F. M., IJzer, S. G., Mashayekhi, K., Haagsman, H. P., & Roelen, B. A.
Affiliations
Utrecht University (NL)
Corresponding author
Roelen, Bernard AJ
ORCID
Venue
BMC Genomics
Publisher
Springer Science and Business Media LLC
Published
2015-04-10
Volume
16
Issue
1
Pages
277-277
ISSN
1471-2164 (ISSN-L)
DOI
10.1186/s12864-015-1448-x
PMC
PMC4397860
OpenAlex
W2048145071
Cited by
26
References
88
Citations by year
2026: 1 · 2022: 6 · 2021: 4 · 2020: 3 · 2019: 2 · 2018: 1 · 2017: 4 · 2016: 4 · 2015: 1
FWCI
1.45
Type
journal article
Language
en
Open access
Licence
Locations
  • BMC Genomics (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (acceptedVersion)
  • Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
Keywords
bovine embryos, pluripotency, inner cell mass, trophectoderm, MAPK inhibition, Nanog, SOX2, Oct4, gene expression profiling, morula, blastocyst, embryonic development, lipid metabolism, RNA splicing
MeSH terms
Animals, Benzamides, Cattle, Cells, Cultured, Diphenylamine, Morula, RNA, Messenger, Gene Expression Regulation, Developmental, Mitogen-Activated Protein Kinase Kinases, Pluripotent Stem Cells, Embryo Culture Techniques, Protein Kinase Inhibitors, Blastocyst Inner Cell Mass

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