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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.


Related in: MedlinePlus

The UNC5C 3’UTR SNP controls its expression level.A. UNC5C expression levels in bovine tissues, as determined via real-time PCR. B. The relative luciferase activity of the 3’UTR region of UNC5C in OV2944-HM-1. The data are presented as the means ± SEM. The p-value was calculated using Student’s t-test. C. The average allele-specific mRNA expression level of UNC5C ± SEM in the heterozygous bovine ovaries based on SNaPshot (n = 5). The ratios of A to G relative to the genomic DNA are shown.
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pone.0131283.g003: The UNC5C 3’UTR SNP controls its expression level.A. UNC5C expression levels in bovine tissues, as determined via real-time PCR. B. The relative luciferase activity of the 3’UTR region of UNC5C in OV2944-HM-1. The data are presented as the means ± SEM. The p-value was calculated using Student’s t-test. C. The average allele-specific mRNA expression level of UNC5C ± SEM in the heterozygous bovine ovaries based on SNaPshot (n = 5). The ratios of A to G relative to the genomic DNA are shown.

Mentions: UNC5C (A+169G) is located in the 3’UTR of UNC5C and may influence the expression level of this gene. Because UNC5C is expressed in several bovine tissues, including the ovary (Fig 3A), we used the murine ovarian tumor line OV2944-HM-1 to assess luciferase activity. Reporters carrying the A allele exhibited higher enzyme activity than those carrying the G allele (Fig 3B). Consistent with the results of the luciferase assay, the level of mRNA generated in the presence of the A allele was higher than that produced in the presence of the G allele according to the allelic mRNA ratio measured in the bovine ovaries (Fig 3C). Consequently, the UNC5C expression level might affect conception rate in cattle.


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)

The UNC5C 3’UTR SNP controls its expression level.A. UNC5C expression levels in bovine tissues, as determined via real-time PCR. B. The relative luciferase activity of the 3’UTR region of UNC5C in OV2944-HM-1. The data are presented as the means ± SEM. The p-value was calculated using Student’s t-test. C. The average allele-specific mRNA expression level of UNC5C ± SEM in the heterozygous bovine ovaries based on SNaPshot (n = 5). The ratios of A to G relative to the genomic DNA are shown.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0131283.g003: The UNC5C 3’UTR SNP controls its expression level.A. UNC5C expression levels in bovine tissues, as determined via real-time PCR. B. The relative luciferase activity of the 3’UTR region of UNC5C in OV2944-HM-1. The data are presented as the means ± SEM. The p-value was calculated using Student’s t-test. C. The average allele-specific mRNA expression level of UNC5C ± SEM in the heterozygous bovine ovaries based on SNaPshot (n = 5). The ratios of A to G relative to the genomic DNA are shown.
Mentions: UNC5C (A+169G) is located in the 3’UTR of UNC5C and may influence the expression level of this gene. Because UNC5C is expressed in several bovine tissues, including the ovary (Fig 3A), we used the murine ovarian tumor line OV2944-HM-1 to assess luciferase activity. Reporters carrying the A allele exhibited higher enzyme activity than those carrying the G allele (Fig 3B). Consistent with the results of the luciferase assay, the level of mRNA generated in the presence of the A allele was higher than that produced in the presence of the G allele according to the allelic mRNA ratio measured in the bovine ovaries (Fig 3C). Consequently, the UNC5C expression level might affect conception rate in cattle.

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.


Related in: MedlinePlus