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Molecular Effects of Polymorphism in the 3'UTR of Unc-5 homolog C Associated with Conception Rate in Holsteins.

Sugimoto M, Gotoh Y, Kawahara T, Sugimoto Y - PLoS ONE (2015)

Bottom Line: To enhance our understanding of the genes involved in conception rates, we conducted a genome-wide association study (GWAS) using 822 Holsteins and identified a single-nucleotide polymorphism (SNP) associated with conception rate: A+169G in the 3' untranslated region (UTR) of unc-5 homolog C (UNC5C).G9a-knockout mice have previously been demonstrated to be subfertile, and we found that UNC5C, G9a, and Netrin-1 expression levels increased from the 4-cell stage to the blastocyst stage in fertilized murine embryos, whereas Rhox5 expression decreased.This study provides novel insights into the role of UNC5C during embryonic development.

View Article: PubMed Central - PubMed

Affiliation: National Livestock Breeding Center, Nishigo, Fukushima, Japan.

ABSTRACT
Conception rates among dairy cows in Japan have declined in recent decades. To enhance our understanding of the genes involved in conception rates, we conducted a genome-wide association study (GWAS) using 822 Holsteins and identified a single-nucleotide polymorphism (SNP) associated with conception rate: A+169G in the 3' untranslated region (UTR) of unc-5 homolog C (UNC5C). Cows with higher conception rates carried the A polymorphism in the UNC5C 3'UTR. Luciferase assays and quantitative analysis of allele ratios revealed that UNC5C transcripts with the A polymorphism were expressed at higher levels than those carrying the G polymorphism. UNC5C transmits either pro- or anti-apoptotic signals depending on the availability of its ligand, Netrin-1. UNC5C expression is negatively regulated by reproductive homeobox X-linked 5 (Rhox5), and the Rhox5 locus is methylated by G9a methyltransferase. G9a-knockout mice have previously been demonstrated to be subfertile, and we found that UNC5C, G9a, and Netrin-1 expression levels increased from the 4-cell stage to the blastocyst stage in fertilized murine embryos, whereas Rhox5 expression decreased. Repression of UNC5C, G9a, or Netrin-1 or forced expression of Rhox5 in the anterior nucleus stage inhibited development to the blastocyst stage, suggesting that cows carrying the G polymorphism in UNC5C might have lower conception rates because of the poor development of preimplantation embryos. This study provides novel insights into the role of UNC5C during embryonic development.

No MeSH data available.


G9a decreases Rhox5 expression.A. Representative images showing UNC5C, Rhox5, G9a, and Netrin-1 (each shown by pink fluorescence) in the anterior nucleus, 2-cell, 4-cell, morula, and blastocyst stages. Bar: 10 μm. B. Representative images showing mCherry (pink fluorescence) in murine embryos at 24 or 96 h after electroporation with mCherry only (Control) or with mCherry and siUNC5C (siUNC5C). Bar: 10 μm. C. Survival rates of murine embryos at 96 h after electroporation with mCherry only (Control), mCherry and siUNC5C (siUNC5C), mCherry and Rhox5 (Rhox5), mCherry and siG9a (siG9a) or mCherry and siNetrin-1 (siNetrin-1).
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pone.0131283.g006: G9a decreases Rhox5 expression.A. Representative images showing UNC5C, Rhox5, G9a, and Netrin-1 (each shown by pink fluorescence) in the anterior nucleus, 2-cell, 4-cell, morula, and blastocyst stages. Bar: 10 μm. B. Representative images showing mCherry (pink fluorescence) in murine embryos at 24 or 96 h after electroporation with mCherry only (Control) or with mCherry and siUNC5C (siUNC5C). Bar: 10 μm. C. Survival rates of murine embryos at 96 h after electroporation with mCherry only (Control), mCherry and siUNC5C (siUNC5C), mCherry and Rhox5 (Rhox5), mCherry and siG9a (siG9a) or mCherry and siNetrin-1 (siNetrin-1).

Mentions: Because G9a-knockout female mice are subfertile [20], G9a, Rhox5, and UNC5C expression might affect fertility. To explore this possibility, we first assessed G9a, Rhox5, and UNC5C expression in fertilized murine embryos. As shown in Fig 6A, the expression levels of UNC5C and G9a increased from the 4-cell stage to the blastocyst stage. However, Rhox5 expression decreased from the 4-cell stage to the blastocyst stage, whereas the expression of Netrin-1, which is a ligand of UNC5C, increased. These distinct expression patterns suggest that the G9a-Rhox5-UNC5C pathway might be important for embryonic development.


Molecular Effects of Polymorphism in the 3'UTR of Unc-5 homolog C Associated with Conception Rate in Holsteins.

Sugimoto M, Gotoh Y, Kawahara T, Sugimoto Y - PLoS ONE (2015)

G9a decreases Rhox5 expression.A. Representative images showing UNC5C, Rhox5, G9a, and Netrin-1 (each shown by pink fluorescence) in the anterior nucleus, 2-cell, 4-cell, morula, and blastocyst stages. Bar: 10 μm. B. Representative images showing mCherry (pink fluorescence) in murine embryos at 24 or 96 h after electroporation with mCherry only (Control) or with mCherry and siUNC5C (siUNC5C). Bar: 10 μm. C. Survival rates of murine embryos at 96 h after electroporation with mCherry only (Control), mCherry and siUNC5C (siUNC5C), mCherry and Rhox5 (Rhox5), mCherry and siG9a (siG9a) or mCherry and siNetrin-1 (siNetrin-1).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493121&req=5

pone.0131283.g006: G9a decreases Rhox5 expression.A. Representative images showing UNC5C, Rhox5, G9a, and Netrin-1 (each shown by pink fluorescence) in the anterior nucleus, 2-cell, 4-cell, morula, and blastocyst stages. Bar: 10 μm. B. Representative images showing mCherry (pink fluorescence) in murine embryos at 24 or 96 h after electroporation with mCherry only (Control) or with mCherry and siUNC5C (siUNC5C). Bar: 10 μm. C. Survival rates of murine embryos at 96 h after electroporation with mCherry only (Control), mCherry and siUNC5C (siUNC5C), mCherry and Rhox5 (Rhox5), mCherry and siG9a (siG9a) or mCherry and siNetrin-1 (siNetrin-1).
Mentions: Because G9a-knockout female mice are subfertile [20], G9a, Rhox5, and UNC5C expression might affect fertility. To explore this possibility, we first assessed G9a, Rhox5, and UNC5C expression in fertilized murine embryos. As shown in Fig 6A, the expression levels of UNC5C and G9a increased from the 4-cell stage to the blastocyst stage. However, Rhox5 expression decreased from the 4-cell stage to the blastocyst stage, whereas the expression of Netrin-1, which is a ligand of UNC5C, increased. These distinct expression patterns suggest that the G9a-Rhox5-UNC5C pathway might be important for embryonic development.

Bottom Line: To enhance our understanding of the genes involved in conception rates, we conducted a genome-wide association study (GWAS) using 822 Holsteins and identified a single-nucleotide polymorphism (SNP) associated with conception rate: A+169G in the 3' untranslated region (UTR) of unc-5 homolog C (UNC5C).G9a-knockout mice have previously been demonstrated to be subfertile, and we found that UNC5C, G9a, and Netrin-1 expression levels increased from the 4-cell stage to the blastocyst stage in fertilized murine embryos, whereas Rhox5 expression decreased.This study provides novel insights into the role of UNC5C during embryonic development.

View Article: PubMed Central - PubMed

Affiliation: National Livestock Breeding Center, Nishigo, Fukushima, Japan.

ABSTRACT
Conception rates among dairy cows in Japan have declined in recent decades. To enhance our understanding of the genes involved in conception rates, we conducted a genome-wide association study (GWAS) using 822 Holsteins and identified a single-nucleotide polymorphism (SNP) associated with conception rate: A+169G in the 3' untranslated region (UTR) of unc-5 homolog C (UNC5C). Cows with higher conception rates carried the A polymorphism in the UNC5C 3'UTR. Luciferase assays and quantitative analysis of allele ratios revealed that UNC5C transcripts with the A polymorphism were expressed at higher levels than those carrying the G polymorphism. UNC5C transmits either pro- or anti-apoptotic signals depending on the availability of its ligand, Netrin-1. UNC5C expression is negatively regulated by reproductive homeobox X-linked 5 (Rhox5), and the Rhox5 locus is methylated by G9a methyltransferase. G9a-knockout mice have previously been demonstrated to be subfertile, and we found that UNC5C, G9a, and Netrin-1 expression levels increased from the 4-cell stage to the blastocyst stage in fertilized murine embryos, whereas Rhox5 expression decreased. Repression of UNC5C, G9a, or Netrin-1 or forced expression of Rhox5 in the anterior nucleus stage inhibited development to the blastocyst stage, suggesting that cows carrying the G polymorphism in UNC5C might have lower conception rates because of the poor development of preimplantation embryos. This study provides novel insights into the role of UNC5C during embryonic development.

No MeSH data available.