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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

Rhox5 decreases UNC5C expression.A. Representative immunoblots with anti-UNC5C, anti-Rhox5, and anti-actin antibodies for OV2944-HM-1 transfected with vector (Control) or Rhox5. B. The relative luciferase activity at the 3’UTR of UNC5C in OV2944-HM-1 co-transfected with vector (Control) or Rhox5. The data are presented as the means ± SEM (n = 6). The p-value was calculated using Student’s t-test.
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pone.0131283.g004: Rhox5 decreases UNC5C expression.A. Representative immunoblots with anti-UNC5C, anti-Rhox5, and anti-actin antibodies for OV2944-HM-1 transfected with vector (Control) or Rhox5. B. The relative luciferase activity at the 3’UTR of UNC5C in OV2944-HM-1 co-transfected with vector (Control) or Rhox5. The data are presented as the means ± SEM (n = 6). The p-value was calculated using Student’s t-test.

Mentions: UNC5C is a proapoptotic molecule [13], and Rhox5 controls its expression in Sertoli cells in the murine testis [20]. To examine whether the Rhox5 expression level affected the UNC5C concentration in the murine ovary, we transfected OV2944-HM-1 with an empty vector or Rhox5 (a Rhox5 expression plasmid). As shown in Fig 4A, increased Rhox5 expression decreased UNC5C expression. Moreover, co-transfection of reporters covering both the 5’ and 3’UTRs of UNC5C with an empty vector or Rhox5 revealed that increased expression of Rhox5 decreased the luciferase activities of both reporters carrying the A and G alleles at the 3’UTR (Fig 4B), suggesting that its expression may affect that of UNC5C.


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)

Rhox5 decreases UNC5C expression.A. Representative immunoblots with anti-UNC5C, anti-Rhox5, and anti-actin antibodies for OV2944-HM-1 transfected with vector (Control) or Rhox5. B. The relative luciferase activity at the 3’UTR of UNC5C in OV2944-HM-1 co-transfected with vector (Control) or Rhox5. The data are presented as the means ± SEM (n = 6). The p-value was calculated using Student’s t-test.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0131283.g004: Rhox5 decreases UNC5C expression.A. Representative immunoblots with anti-UNC5C, anti-Rhox5, and anti-actin antibodies for OV2944-HM-1 transfected with vector (Control) or Rhox5. B. The relative luciferase activity at the 3’UTR of UNC5C in OV2944-HM-1 co-transfected with vector (Control) or Rhox5. The data are presented as the means ± SEM (n = 6). The p-value was calculated using Student’s t-test.
Mentions: UNC5C is a proapoptotic molecule [13], and Rhox5 controls its expression in Sertoli cells in the murine testis [20]. To examine whether the Rhox5 expression level affected the UNC5C concentration in the murine ovary, we transfected OV2944-HM-1 with an empty vector or Rhox5 (a Rhox5 expression plasmid). As shown in Fig 4A, increased Rhox5 expression decreased UNC5C expression. Moreover, co-transfection of reporters covering both the 5’ and 3’UTRs of UNC5C with an empty vector or Rhox5 revealed that increased expression of Rhox5 decreased the luciferase activities of both reporters carrying the A and G alleles at the 3’UTR (Fig 4B), suggesting that its expression may affect that of UNC5C.

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