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A missense mutation in solute carrier family 12, member 1 ( SLC12A1 ) causes hydrallantois in Japanese Black cattle

View Article: PubMed Central - PubMed

ABSTRACT

Background: Hydrallantois is the excessive accumulation of fluid within the allantoic cavity in pregnant animals and is associated with fetal mortality. Although the incidence of hydrallantois is very low in artificial insemination breeding programs in cattle, recently 38 cows with the phenotypic appearance of hydrallantois were reported in a local subpopulation of Japanese Black cattle. Of these, 33 were traced back to the same sire; however, both their parents were reported healthy, suggesting that hydrallantois is a recessive inherited disorder. To identify autozygous chromosome segments shared by individuals with hydrallantois and the causative mutation in Japanese Black cattle, we performed autozygosity mapping using single-nucleotide polymorphism (SNP) array and exome sequencing.

Results: Shared haplotypes of the affected fetuses spanned 3.52 Mb on bovine chromosome 10. Exome sequencing identified a SNP (g.62382825G > A, p.Pro372Leu) in exon 10 of solute carrier family 12, member 1 (SLC12A1), the genotype of which was compatible with recessive inheritance. SLC12A1 serves as a reabsorption molecule of Na+-K+-2Cl− in the apical membrane of the thick ascending limb of the loop of Henle in the kidney. We observed that the concentration of Na+-Cl− increased in allantoic fluid of homozygous SLC12A1 (g.62382825G > A) in a hydrallantois individual. In addition, SLC12A1-positive signals were localized at the apical membrane in the kidneys of unaffected fetuses, whereas they were absent from the apical membrane in the kidneys of affected fetuses. These results suggested that p.Pro372Leu affects the membrane localization of SLC12A1, and in turn, may impair its transporter activity. Surveillance of the risk-allele frequency revealed that the carriers were restricted to the local subpopulation of Japanese Black cattle. Moreover, we identified a founder individual that carried the mutation (g.62382825G > A).

Conclusions: In this study, we mapped the shared haplotypes of affected fetuses using autozygosity mapping and identified a de novo mutation in the SLC12A1 gene that was associated with hydrallantois in Japanese Black cattle. In kidneys of hydrallantois-affected fetuses, the mutation in SLC12A1 impaired the apical membrane localization of SLC12A1 and reabsorption of Na+-K+-2Cl− in the thick ascending limb of the loop of Henle, leading to a defect in the concentration of urine via the countercurrent mechanism. Consequently, the affected fetuses exhibited polyuria that accumulated in the allantoic cavity. Surveillance of the risk-allele frequency indicated that carriers were not widespread throughout the Japanese Black cattle population. Moreover, we identified the founder individual, and thus could effectively manage the disorder in the population.

Electronic supplementary material: The online version of this article (doi:10.1186/s12864-016-3035-1) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus

Identification of a founder individual for hydrallantois in Japanese Black cattle. gDNA was extracted from semen (germ-line cells) from sire_X, the father, maternal grandsire, maternal great-grandsire, and from endometrial cells (somatic cells) derived from the uterus of the mother of sire_X. The gDNA was genotyped using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. a, b Non-risk-haplotypes/non-risk-allele carriers, and risk-haplotype/risk-allele carriers are represented with blue-filled symbols and magenta-filled symbols, respectively. Individuals for which we could not determine the haplotype or the allele by genotyping are denoted by empty symbols (ND). c Calibration curve of g.62382825G > A. d Sequence electropherograms G/A allele molar ratio in gDNA from semen (germ-line cells, d2) and gDNA from dermal fibroblast (somatic cells, d3) from sire_X was calculated with the ratio of the peak height of G:A alleles in gDNA of the calibration sample (d1)
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Fig6: Identification of a founder individual for hydrallantois in Japanese Black cattle. gDNA was extracted from semen (germ-line cells) from sire_X, the father, maternal grandsire, maternal great-grandsire, and from endometrial cells (somatic cells) derived from the uterus of the mother of sire_X. The gDNA was genotyped using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. a, b Non-risk-haplotypes/non-risk-allele carriers, and risk-haplotype/risk-allele carriers are represented with blue-filled symbols and magenta-filled symbols, respectively. Individuals for which we could not determine the haplotype or the allele by genotyping are denoted by empty symbols (ND). c Calibration curve of g.62382825G > A. d Sequence electropherograms G/A allele molar ratio in gDNA from semen (germ-line cells, d2) and gDNA from dermal fibroblast (somatic cells, d3) from sire_X was calculated with the ratio of the peak height of G:A alleles in gDNA of the calibration sample (d1)

Mentions: The most effective way to eliminate the causative mutations from a population is to identify a founder individual and then perform targeted genotyping of the related sires used for breeding. Pedigree analysis showed that parents of the nine affected fetuses traced back to sire_X (Fig. 2). To trace back the risk-haplotype and the risk-allele of sire_X, we genotyped the parents, the maternal grandsire, and the maternal great-grandsire of sire_X (Fig. 6a, b) using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. gDNA was extracted from the semen (germ-line cells) of sire_X, the father, the maternal grandsire, the maternal great-grandsire, and from the endometrial cells (somatic cells) derived from the uterus of the mother of sire_X (Fig. 6a, b). We found that the mother of sire_X was heterozygous for the risk-haplotype, whereas the father, maternal grandsire, and maternal great-grandsire did not have the risk-haplotype (Fig. 6a), indicating that the risk-haplotype was transmitted from the mother. However, the mother did not have the risk-allele (Fig. 6b), suggesting that a de novo mutation was generated in the germ-line cells of the mother or in the germ-line cells of sire_X with mosaic status.Fig. 6


A missense mutation in solute carrier family 12, member 1 ( SLC12A1 ) causes hydrallantois in Japanese Black cattle
Identification of a founder individual for hydrallantois in Japanese Black cattle. gDNA was extracted from semen (germ-line cells) from sire_X, the father, maternal grandsire, maternal great-grandsire, and from endometrial cells (somatic cells) derived from the uterus of the mother of sire_X. The gDNA was genotyped using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. a, b Non-risk-haplotypes/non-risk-allele carriers, and risk-haplotype/risk-allele carriers are represented with blue-filled symbols and magenta-filled symbols, respectively. Individuals for which we could not determine the haplotype or the allele by genotyping are denoted by empty symbols (ND). c Calibration curve of g.62382825G > A. d Sequence electropherograms G/A allele molar ratio in gDNA from semen (germ-line cells, d2) and gDNA from dermal fibroblast (somatic cells, d3) from sire_X was calculated with the ratio of the peak height of G:A alleles in gDNA of the calibration sample (d1)
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Related In: Results  -  Collection

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Fig6: Identification of a founder individual for hydrallantois in Japanese Black cattle. gDNA was extracted from semen (germ-line cells) from sire_X, the father, maternal grandsire, maternal great-grandsire, and from endometrial cells (somatic cells) derived from the uterus of the mother of sire_X. The gDNA was genotyped using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. a, b Non-risk-haplotypes/non-risk-allele carriers, and risk-haplotype/risk-allele carriers are represented with blue-filled symbols and magenta-filled symbols, respectively. Individuals for which we could not determine the haplotype or the allele by genotyping are denoted by empty symbols (ND). c Calibration curve of g.62382825G > A. d Sequence electropherograms G/A allele molar ratio in gDNA from semen (germ-line cells, d2) and gDNA from dermal fibroblast (somatic cells, d3) from sire_X was calculated with the ratio of the peak height of G:A alleles in gDNA of the calibration sample (d1)
Mentions: The most effective way to eliminate the causative mutations from a population is to identify a founder individual and then perform targeted genotyping of the related sires used for breeding. Pedigree analysis showed that parents of the nine affected fetuses traced back to sire_X (Fig. 2). To trace back the risk-haplotype and the risk-allele of sire_X, we genotyped the parents, the maternal grandsire, and the maternal great-grandsire of sire_X (Fig. 6a, b) using GGP Bovine 26 K BeadChip array and g.62382825G > A using PCR. gDNA was extracted from the semen (germ-line cells) of sire_X, the father, the maternal grandsire, the maternal great-grandsire, and from the endometrial cells (somatic cells) derived from the uterus of the mother of sire_X (Fig. 6a, b). We found that the mother of sire_X was heterozygous for the risk-haplotype, whereas the father, maternal grandsire, and maternal great-grandsire did not have the risk-haplotype (Fig. 6a), indicating that the risk-haplotype was transmitted from the mother. However, the mother did not have the risk-allele (Fig. 6b), suggesting that a de novo mutation was generated in the germ-line cells of the mother or in the germ-line cells of sire_X with mosaic status.Fig. 6

View Article: PubMed Central - PubMed

ABSTRACT

Background: Hydrallantois is the excessive accumulation of fluid within the allantoic cavity in pregnant animals and is associated with fetal mortality. Although the incidence of hydrallantois is very low in artificial insemination breeding programs in cattle, recently 38 cows with the phenotypic appearance of hydrallantois were reported in a local subpopulation of Japanese Black cattle. Of these, 33 were traced back to the same sire; however, both their parents were reported healthy, suggesting that hydrallantois is a recessive inherited disorder. To identify autozygous chromosome segments shared by individuals with hydrallantois and the causative mutation in Japanese Black cattle, we performed autozygosity mapping using single-nucleotide polymorphism (SNP) array and exome sequencing.

Results: Shared haplotypes of the affected fetuses spanned 3.52 Mb on bovine chromosome 10. Exome sequencing identified a SNP (g.62382825G > A, p.Pro372Leu) in exon 10 of solute carrier family 12, member 1 (SLC12A1), the genotype of which was compatible with recessive inheritance. SLC12A1 serves as a reabsorption molecule of Na+-K+-2Cl− in the apical membrane of the thick ascending limb of the loop of Henle in the kidney. We observed that the concentration of Na+-Cl− increased in allantoic fluid of homozygous SLC12A1 (g.62382825G > A) in a hydrallantois individual. In addition, SLC12A1-positive signals were localized at the apical membrane in the kidneys of unaffected fetuses, whereas they were absent from the apical membrane in the kidneys of affected fetuses. These results suggested that p.Pro372Leu affects the membrane localization of SLC12A1, and in turn, may impair its transporter activity. Surveillance of the risk-allele frequency revealed that the carriers were restricted to the local subpopulation of Japanese Black cattle. Moreover, we identified a founder individual that carried the mutation (g.62382825G > A).

Conclusions: In this study, we mapped the shared haplotypes of affected fetuses using autozygosity mapping and identified a de novo mutation in the SLC12A1 gene that was associated with hydrallantois in Japanese Black cattle. In kidneys of hydrallantois-affected fetuses, the mutation in SLC12A1 impaired the apical membrane localization of SLC12A1 and reabsorption of Na+-K+-2Cl− in the thick ascending limb of the loop of Henle, leading to a defect in the concentration of urine via the countercurrent mechanism. Consequently, the affected fetuses exhibited polyuria that accumulated in the allantoic cavity. Surveillance of the risk-allele frequency indicated that carriers were not widespread throughout the Japanese Black cattle population. Moreover, we identified the founder individual, and thus could effectively manage the disorder in the population.

Electronic supplementary material: The online version of this article (doi:10.1186/s12864-016-3035-1) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus