Regulation of gene expression in the bovine blastocyst by colony stimulating factor 2

Abstract

BackgroundColony stimulating factor 2 can have multiple effects on the function of the preimplantation embryo that include increased potential to develop to the blastocyst stage, reduced apoptosis, and enhanced ability of inner cell mass (ICM) to remain pluripotent after culture. The objective of the current experiment was to identify genes regulated by CSF2 in the ICM and trophectoderm (TE) of the bovine blastocyst with the goal of identifying possible molecular pathways by which CSF2 increases developmental competence for survival. Embryos were produced in vitro and cultured from Day 6 to 8 in serum-free medium containing 10 ng/ml recombinant bovine CSF2 or vehicle. Blastocysts were harvested at Day 8 and ICM separated from TE by magnetic-activated cell sorting. RNA was purified and used to prepare amplified cDNA, which was then subjected to high-throughput sequencing using the SOLiD 4.0 system. Three pools of amplified cDNA were analyzed per treatment.ResultsThe number of genes whose expression was regulated by CSF2, using P < 0.05 and >1.5-fold difference as cut-offs, was 945 in the ICM (242 upregulated by CSF2 and 703 downregulated) and 886 in the TE (401 upregulated by CSF2 and 485 downregulated). Only 49 genes were regulated in a similar manner by CSF2 in both cell types. The three significant annotation clusters in which genes regulated by ICM were overrepresented were related to membrane signaling. Genes downregulated by CSF2 in ICM were overrepresented in several pathways including those for ERK and AKT signaling. The only significant annotation cluster containing an overrepresentation of genes regulated by CSF2 in TE was for secreted or extracellular proteins. In addition, genes downregulated in TE were overrepresented in TGFβ and Nanog pathways.ConclusionsDifferentiation of the blastocyst is such that, by Day 8 after fertilization, the ICM and TE respond differently to CSF2. Analysis of the genes regulated by CSF2 in ICM and TE are suggestive that CSF2 reinforces developmental fate and function of both cell lineages.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-2038-y) contains supplementary material, which is available to authorized users.

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“…Other potentially relevant downregulated genes encode transcripts associated with Alzheimer's disease (Sorl1) (Nicolas et al, 2016), leukodystrophy (Abcd1 and its downstream mediator of pathology, Ch25h) (Gong et al, 2017;Jang et al, 2016), and the CSF-2 target gene Pkch, encoding protein kinase Ch, a regulator of lipid metabolism and suppressor of nitrous oxide (NO) production by macrophages (Torisu et al, 2016;Ozawa et al, 2016; Figure 5B).…”

Genome-wide association analysis uncovers variants for reproductive variation across dog breeds and links to domestication

“…Additionally, CACNA2D3 is one of four genes recently described as influencing cranial morphology in human populations [94]. In other domesticated animals, CACNA2D3 is downregulated by Colony Stimulating Factor 2 ( CSF2 ) in the trophectoderm of pregnant cattle, which increases the ability of the preimplantation embryo to advance to the blastocyst stage [95]. In the closely related wolf, CACNA2D3 is under diversifying selection associated with environmental adaptations to altitude [96–98].…”

Genome wide association analysis identifies genetic variants associated with reproductive variation across domestic dog breeds and uncovers links to domestication

“…Additionally, CACNA2D3 is one of four genes recently described as influencing cranial morphology in human populations (Paternoster et al 2012). In other domesticated animals, CACNA2D3 is downregulated by Colony Stimulating Factor 2 ( CSF2 ) in the trophoectoderm of pregnant cattle, which increases the ability of the preimplantation embryo to advance to the blastocyst stage (Ozawa et al 2016). In the closely related wolf, CACNA2D3 is under diversifying selection associated with environmental adaptations to altitude (Pilot et al 2014, Schweizer et al 2016, Zhang et al 2014).…”