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Detection of differentially expressed genes between Erhualian and Large White placentas on day 75 and 90 of gestation.

Zhou QY, Fang MD, Huang TH, Li CC, Yu M, Zhao SH - BMC Genomics (2009)

Bottom Line: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality.Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds.Differential expression of six candidate imprinted genes was also confirmed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, PR China. zhouquanyong@webmail.hzau.edu.cn

ABSTRACT

Background: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality. Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds. To understand the molecular basis of placental development during late gestation in Chinese indigenous and Western breeds with different placental efficiency, female placental samples were collected from six pregnant Erhualian gilts at gestation day 75 (E75) and day 90 (E90) and from six pregnant Large White gilts at gestation day 75 (L75) and day 90 (L90). Two female placentas from one sow were used to extract RNA and then pooled in equal volumes. Twelve pooled samples were hybridized to the porcine Affymetrix GeneChip.

Results: A total of 226 and 577 transcripts were detected that were differentially expressed between E75 and L75 and between E90 and L90 (p < 0.01, q < 0.2), respectively. Gene Ontology (GO) analysis revealed that these genes belong to the class of genes that participate in angiogenesis and development. Real-time RT-PCR confirmed the differential expression of eight selected genes. Significant differential expression of five genes in the VEGF pathway was also detected between the breeds. A search of chromosomal location revealed that 44 differentially expressed genes located to QTL regions related to reproduction. Differential expression of six candidate imprinted genes was also confirmed. Three of the six genes (PLAGL1, DIRAS3, and SLC38A4) showed monoallelic expression in the porcine placenta.

Conclusion: Our study detected many genes that showed differential expression between placentas of two divergent breed of pigs, and confirmed the imprinting of three genes. These findings help to elucidate the genetic control of placental efficiency and improve the understanding of placental development.

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Imprinting status of PLAGL1, SLC38A4 and DIRAS3 genes in porcine placentas. The results indicated that PLAGL1 and SLC38A4 genes are paternal expression in porcine placenta. DIRAS3 gene is monoallelic expression in porcine placenta.
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Figure 4: Imprinting status of PLAGL1, SLC38A4 and DIRAS3 genes in porcine placentas. The results indicated that PLAGL1 and SLC38A4 genes are paternal expression in porcine placenta. DIRAS3 gene is monoallelic expression in porcine placenta.

Mentions: To further investigate the allelic expression of candidate imprinted genes in the porcine placenta, cDNA sequences of from both breeds were compared. In the PLAGL1 gene, one single nucleotide polymorphism (SNP) (T/C) was detected at position 1428 (FJ746562). In the SLC38A4 gene, an SNP (C/T) was detected at position 21 (FJ746563), and in the DIRAS3 gene, one SNP (T/G) was revealed at position 428 (NM_001044598). Restriction enzyme digestion (TaqI, Btg I, and Apek I) were used to digest the genomic DNA and cDNA of placentas. At least three heterozygotes were used to confirm the expression of imprinted genes, and the results showed that PLAGL1 and SLC38A4 were paternal expressed. Unfortunately, we could not detect the parental source of DIRAS3 gene, because that maternal genetype also was heterozygotes (Figure 4). No cSNPs were detected in other candidate imprinted genes, thus the imprinting status was not determined for the remaining genes.


Detection of differentially expressed genes between Erhualian and Large White placentas on day 75 and 90 of gestation.

Zhou QY, Fang MD, Huang TH, Li CC, Yu M, Zhao SH - BMC Genomics (2009)

Imprinting status of PLAGL1, SLC38A4 and DIRAS3 genes in porcine placentas. The results indicated that PLAGL1 and SLC38A4 genes are paternal expression in porcine placenta. DIRAS3 gene is monoallelic expression in porcine placenta.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2724418&req=5

Figure 4: Imprinting status of PLAGL1, SLC38A4 and DIRAS3 genes in porcine placentas. The results indicated that PLAGL1 and SLC38A4 genes are paternal expression in porcine placenta. DIRAS3 gene is monoallelic expression in porcine placenta.
Mentions: To further investigate the allelic expression of candidate imprinted genes in the porcine placenta, cDNA sequences of from both breeds were compared. In the PLAGL1 gene, one single nucleotide polymorphism (SNP) (T/C) was detected at position 1428 (FJ746562). In the SLC38A4 gene, an SNP (C/T) was detected at position 21 (FJ746563), and in the DIRAS3 gene, one SNP (T/G) was revealed at position 428 (NM_001044598). Restriction enzyme digestion (TaqI, Btg I, and Apek I) were used to digest the genomic DNA and cDNA of placentas. At least three heterozygotes were used to confirm the expression of imprinted genes, and the results showed that PLAGL1 and SLC38A4 were paternal expressed. Unfortunately, we could not detect the parental source of DIRAS3 gene, because that maternal genetype also was heterozygotes (Figure 4). No cSNPs were detected in other candidate imprinted genes, thus the imprinting status was not determined for the remaining genes.

Bottom Line: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality.Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds.Differential expression of six candidate imprinted genes was also confirmed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, PR China. zhouquanyong@webmail.hzau.edu.cn

ABSTRACT

Background: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality. Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds. To understand the molecular basis of placental development during late gestation in Chinese indigenous and Western breeds with different placental efficiency, female placental samples were collected from six pregnant Erhualian gilts at gestation day 75 (E75) and day 90 (E90) and from six pregnant Large White gilts at gestation day 75 (L75) and day 90 (L90). Two female placentas from one sow were used to extract RNA and then pooled in equal volumes. Twelve pooled samples were hybridized to the porcine Affymetrix GeneChip.

Results: A total of 226 and 577 transcripts were detected that were differentially expressed between E75 and L75 and between E90 and L90 (p < 0.01, q < 0.2), respectively. Gene Ontology (GO) analysis revealed that these genes belong to the class of genes that participate in angiogenesis and development. Real-time RT-PCR confirmed the differential expression of eight selected genes. Significant differential expression of five genes in the VEGF pathway was also detected between the breeds. A search of chromosomal location revealed that 44 differentially expressed genes located to QTL regions related to reproduction. Differential expression of six candidate imprinted genes was also confirmed. Three of the six genes (PLAGL1, DIRAS3, and SLC38A4) showed monoallelic expression in the porcine placenta.

Conclusion: Our study detected many genes that showed differential expression between placentas of two divergent breed of pigs, and confirmed the imprinting of three genes. These findings help to elucidate the genetic control of placental efficiency and improve the understanding of placental development.

Show MeSH